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(-) Remove <label class='research-domain' title='Cellular and Developmental Biology'>LS3 (140)</label> filter LS3 (140)
Project acronym ACTOMYO
Project Mechanisms of actomyosin-based contractility during cytokinesis
Researcher (PI) Ana Costa Xavier de Carvalho
Host Institution (HI) INSTITUTO DE BIOLOGIA MOLECULAR E CELULAR-IBMC
Summary Cytokinesis completes cell division by partitioning the contents of the mother cell to the two daughter cells. This process is accomplished through the assembly and constriction of a contractile ring, a complex actomyosin network that remains poorly understood on the molecular level. Research in cytokinesis has overwhelmingly focused on signaling mechanisms that dictate when and where the contractile ring is assembled. By contrast, the research I propose here addresses fundamental questions about the structural and functional properties of the contractile ring itself. We will use the nematode C. elegans to exploit the power of quantitative live imaging assays in an experimentally tractable metazoan organism. The early C. elegans embryo is uniquely suited to the study of the contractile ring, as cells dividing perpendicularly to the imaging plane provide a full end-on view of the contractile ring throughout constriction. This greatly facilitates accurate measurements of constriction kinetics, ring width and thickness, and levels as well as dynamics of fluorescently-tagged contractile ring components. Combining image-based assays with powerful molecular replacement technology for structure-function studies, we will 1) determine the contribution of branched and non-branched actin filament populations to contractile ring formation; 2) explore its ultra-structural organization in collaboration with a world expert in electron microcopy; 3) investigate how the contractile ring network is dynamically remodeled during constriction with the help of a novel laser microsurgery assay that has uncovered a remarkably robust ring repair mechanism; and 4) use a targeted RNAi screen and phenotype profiling to identify new components of actomyosin contractile networks. The results from this interdisciplinary project will significantly enhance our mechanistic understanding of cytokinesis and other cellular processes that involve actomyosin-based contractility.
Cytokinesis completes cell division by partitioning the contents of the mother cell to the two daughter cells. This process is accomplished through the assembly and constriction of a contractile ring, a complex actomyosin network that remains poorly understood on the molecular level. Research in cytokinesis has overwhelmingly focused on signaling mechanisms that dictate when and where the contractile ring is assembled. By contrast, the research I propose here addresses fundamental questions about the structural and functional properties of the contractile ring itself. We will use the nematode C. elegans to exploit the power of quantitative live imaging assays in an experimentally tractable metazoan organism. The early C. elegans embryo is uniquely suited to the study of the contractile ring, as cells dividing perpendicularly to the imaging plane provide a full end-on view of the contractile ring throughout constriction. This greatly facilitates accurate measurements of constriction kinetics, ring width and thickness, and levels as well as dynamics of fluorescently-tagged contractile ring components. Combining image-based assays with powerful molecular replacement technology for structure-function studies, we will 1) determine the contribution of branched and non-branched actin filament populations to contractile ring formation; 2) explore its ultra-structural organization in collaboration with a world expert in electron microcopy; 3) investigate how the contractile ring network is dynamically remodeled during constriction with the help of a novel laser microsurgery assay that has uncovered a remarkably robust ring repair mechanism; and 4) use a targeted RNAi screen and phenotype profiling to identify new components of actomyosin contractile networks. The results from this interdisciplinary project will significantly enhance our mechanistic understanding of cytokinesis and other cellular processes that involve actomyosin-based contractility.
Project acronym APOQUANT
Project The quantitative Bcl-2 interactome in apoptosis: decoding how cancer cells escape death
Researcher (PI) Ana Jesús García Sáez
Host Institution (HI) EBERHARD KARLS UNIVERSITAET TUEBINGEN
Summary The proteins of the Bcl-2 family function as key regulators of apoptosis by controlling the permeabilization of the mitochondrial outer membrane. They form an intricate, fine-tuned interaction network which is altered in cancer cells to avoid cell death. Currently, we do not understand how signaling within this network, which combines events in cytosol and membranes, is orchestrated to decide the cell fate. The main goal of this proposal is to unravel how apoptosis signaling is integrated by the Bcl-2 network by determining the quantitative Bcl-2 interactome and building with it a mathematical model that identifies which interactions determine the overall outcome. To this aim, we have established a reconstituted system for the quantification of the interactions between Bcl-2 proteins not only in solution but also in membranes at the single molecule level by fluorescence correlation spectroscopy (FCS). (1) This project aims to quantify the relative affinities between an reconstituted Bcl-2 network by FCS. (2) This will be combined with quantitative studies in living cells, which include the signaling pathway in its entirety. To this aim, we will develop new FCS methods for mitochondria. (3) The structural and dynamic aspects of the Bcl-2 network will be studied by super resolution and live cell microscopy. (4) The acquired knowledge will be used to build a mathematical model that uncovers how the multiple interactions within the Bcl-2 network are integrated and identifies critical steps in apoptosis regulation. These studies are expected to broaden the general knowledge about the design principles of cellular signaling as well as how cancer cells alter the Bcl-2 network to escape cell death. This systems analysis will allow us to predict which perturbations in the Bcl-2 network of cancer cells can switch signaling towards cell death. Ultimately it could be translated into clinical applications for anticancer therapy.
The proteins of the Bcl-2 family function as key regulators of apoptosis by controlling the permeabilization of the mitochondrial outer membrane. They form an intricate, fine-tuned interaction network which is altered in cancer cells to avoid cell death. Currently, we do not understand how signaling within this network, which combines events in cytosol and membranes, is orchestrated to decide the cell fate. The main goal of this proposal is to unravel how apoptosis signaling is integrated by the Bcl-2 network by determining the quantitative Bcl-2 interactome and building with it a mathematical model that identifies which interactions determine the overall outcome. To this aim, we have established a reconstituted system for the quantification of the interactions between Bcl-2 proteins not only in solution but also in membranes at the single molecule level by fluorescence correlation spectroscopy (FCS). (1) This project aims to quantify the relative affinities between an reconstituted Bcl-2 network by FCS. (2) This will be combined with quantitative studies in living cells, which include the signaling pathway in its entirety. To this aim, we will develop new FCS methods for mitochondria. (3) The structural and dynamic aspects of the Bcl-2 network will be studied by super resolution and live cell microscopy. (4) The acquired knowledge will be used to build a mathematical model that uncovers how the multiple interactions within the Bcl-2 network are integrated and identifies critical steps in apoptosis regulation. These studies are expected to broaden the general knowledge about the design principles of cellular signaling as well as how cancer cells alter the Bcl-2 network to escape cell death. This systems analysis will allow us to predict which perturbations in the Bcl-2 network of cancer cells can switch signaling towards cell death. Ultimately it could be translated into clinical applications for anticancer therapy.
Project acronym APPL
Project Anionic PhosPhoLipids in plant receptor kinase signaling
Researcher (PI) Yvon Jaillais
Summary "In plants, receptor kinases form the largest family of plasma membrane (PM) receptors and they are involved in virtually all aspects of the plant life, including development, immunity and reproduction. In animals, key molecules that orchestrate the recruitment of signaling proteins to membranes are anionic phospholipids (e.g. phosphatidylinositol phosphate or PIPs). Besides, recent reports in animal and yeast cells suggest the existence of PM nanodomains that are independent of cholesterol and lipid phase and rely on anionic phospholipids as well as electrostatic protein/lipid interactions. Strikingly, we know very little on the role of anionic phospholipids in plant signaling. However, our preliminary data suggest that BKI1, an inhibitory protein of the steroid receptor kinase BRI1, interacts with various PIPs in vitro and is likely targeted to the PM by electrostatic interactions with these anionic lipids. These results open the possibility that BRI1, but also other receptor kinases, might be regulated by anionic phospholipids in plants. Here, we propose to analyze the function of anionic phospholipids in BRI1 signaling, using the root epidermis as a model system. First, we will ask what are the lipids that control membrane surface charge in this tissue and recruit BR-signaling component to the PM. Second, we will probe the presence of PIP-enriched nanodomains at the plant PM using super-resolution microscopy techniques and investigate the roles of these domains in BRI1 signaling. Finally, we will analyze the function of the BKI1-related plant-specific family of anionic phospholipid effectors in plant development. In summary, using a transversal approach ranging from in vitro studies to in vivo validation and whole organism physiology, this work will unravel the interplay between anionic phospholipids and receptor signaling in plants."
"In plants, receptor kinases form the largest family of plasma membrane (PM) receptors and they are involved in virtually all aspects of the plant life, including development, immunity and reproduction. In animals, key molecules that orchestrate the recruitment of signaling proteins to membranes are anionic phospholipids (e.g. phosphatidylinositol phosphate or PIPs). Besides, recent reports in animal and yeast cells suggest the existence of PM nanodomains that are independent of cholesterol and lipid phase and rely on anionic phospholipids as well as electrostatic protein/lipid interactions. Strikingly, we know very little on the role of anionic phospholipids in plant signaling. However, our preliminary data suggest that BKI1, an inhibitory protein of the steroid receptor kinase BRI1, interacts with various PIPs in vitro and is likely targeted to the PM by electrostatic interactions with these anionic lipids. These results open the possibility that BRI1, but also other receptor kinases, might be regulated by anionic phospholipids in plants. Here, we propose to analyze the function of anionic phospholipids in BRI1 signaling, using the root epidermis as a model system. First, we will ask what are the lipids that control membrane surface charge in this tissue and recruit BR-signaling component to the PM. Second, we will probe the presence of PIP-enriched nanodomains at the plant PM using super-resolution microscopy techniques and investigate the roles of these domains in BRI1 signaling. Finally, we will analyze the function of the BKI1-related plant-specific family of anionic phospholipid effectors in plant development. In summary, using a transversal approach ranging from in vitro studies to in vivo validation and whole organism physiology, this work will unravel the interplay between anionic phospholipids and receptor signaling in plants."
Project acronym ASYMMEM
Project Lipid asymmetry: a cellular battery?
Researcher (PI) André NADLER
Summary It is a basic textbook notion that the plasma membranes of virtually all organisms display an asymmetric lipid distribution between inner and outer leaflets far removed from thermodynamic equilibrium. As a fundamental biological principle, lipid asymmetry has been linked to numerous cellular processes. However, a clear mechanistic justification for the continued existence of lipid asymmetry throughout evolution has yet to be established. We propose here that lipid asymmetry serves as a store of potential energy that is used to fuel energy-intense membrane remodelling and signalling events for instance during membrane fusion and fission. This implies that rapid, local changes of trans-membrane lipid distribution rather than a continuously maintained out-of-equilibrium situation are crucial for cellular function. Consequently, new methods for quantifying the kinetics of lipid trans-bilayer movement are required, as traditional approaches are mostly suited for analysing quasi-steady-state conditions. Addressing this need, we will develop and employ novel photochemical lipid probes and lipid biosensors to quantify localized trans-bilayer lipid movement. We will use these tools for identifying yet unknown protein components of the lipid asymmetry regulating machinery and analyse their function with regard to membrane dynamics and signalling in cell motility. Focussing on cell motility enables targeted chemical and genetic perturbations while monitoring lipid dynamics on timescales and in membrane structures that are well suited for light microscopy. Ultimately, we aim to reconstitute lipid asymmetry as a driving force for membrane remodelling in vitro. We expect that our work will break new ground in explaining one of the least understood features of the plasma membrane and pave the way for a new, dynamic membrane model. Since the plasma membrane serves as the major signalling hub, this will have impact in almost every area of the life sciences.
It is a basic textbook notion that the plasma membranes of virtually all organisms display an asymmetric lipid distribution between inner and outer leaflets far removed from thermodynamic equilibrium. As a fundamental biological principle, lipid asymmetry has been linked to numerous cellular processes. However, a clear mechanistic justification for the continued existence of lipid asymmetry throughout evolution has yet to be established. We propose here that lipid asymmetry serves as a store of potential energy that is used to fuel energy-intense membrane remodelling and signalling events for instance during membrane fusion and fission. This implies that rapid, local changes of trans-membrane lipid distribution rather than a continuously maintained out-of-equilibrium situation are crucial for cellular function. Consequently, new methods for quantifying the kinetics of lipid trans-bilayer movement are required, as traditional approaches are mostly suited for analysing quasi-steady-state conditions. Addressing this need, we will develop and employ novel photochemical lipid probes and lipid biosensors to quantify localized trans-bilayer lipid movement. We will use these tools for identifying yet unknown protein components of the lipid asymmetry regulating machinery and analyse their function with regard to membrane dynamics and signalling in cell motility. Focussing on cell motility enables targeted chemical and genetic perturbations while monitoring lipid dynamics on timescales and in membrane structures that are well suited for light microscopy. Ultimately, we aim to reconstitute lipid asymmetry as a driving force for membrane remodelling in vitro. We expect that our work will break new ground in explaining one of the least understood features of the plasma membrane and pave the way for a new, dynamic membrane model. Since the plasma membrane serves as the major signalling hub, this will have impact in almost every area of the life sciences.
Project acronym AuxinER
Project Mechanisms of Auxin-dependent Signaling in the Endoplasmic Reticulum
Researcher (PI) Jürgen Kleine-Vehn
Host Institution (HI) UNIVERSITAET FUER BODENKULTUR WIEN
Summary The phytohormone auxin has profound importance for plant development. The extracellular AUXIN BINDING PROTEIN1 (ABP1) and the nuclear AUXIN F-BOX PROTEINs (TIR1/AFBs) auxin receptors perceive fast, non-genomic and slow, genomic auxin responses, respectively. Despite the fact that ABP1 mainly localizes to the endoplasmic reticulum (ER), until now it has been proposed to be active only in the extracellular matrix (reviewed in Sauer and Kleine-Vehn, 2011). Just recently, ABP1 function was also linked to genomic responses, modulating TIR1/AFB-dependent processes (Tromas et al., 2013). Intriguingly, the genomic effect of ABP1 appears to be at least partially independent of the endogenous auxin indole 3-acetic acid (IAA) (Paque et al., 2014). In this proposal my main research objective is to unravel the importance of the ER for genomic auxin responses. The PIN-LIKES (PILS) putative carriers for auxinic compounds also localize to the ER and determine the cellular sensitivity to auxin. PILS5 gain-of-function reduces canonical auxin signaling (Barbez et al., 2012) and phenocopies abp1 knock down lines (Barbez et al., 2012, Paque et al., 2014). Accordingly, a PILS-dependent substrate could be a negative regulator of ABP1 function in the ER. Based on our unpublished data, an IAA metabolite could play a role in ABP1-dependent processes in the ER, possibly providing feedback on the canonical nuclear IAA-signaling. I hypothesize that the genomic auxin response may be an integration of auxin- and auxin-metabolite-dependent nuclear and ER localized signaling, respectively. This proposed project aims to characterize a novel auxin-signaling paradigm in plants. We will employ state of the art interdisciplinary (biochemical, biophysical, computational modeling, molecular, and genetic) methods to assess the projected research. The identification of the proposed auxin conjugate-dependent signal could have far reaching plant developmental and biotechnological importance.
The phytohormone auxin has profound importance for plant development. The extracellular AUXIN BINDING PROTEIN1 (ABP1) and the nuclear AUXIN F-BOX PROTEINs (TIR1/AFBs) auxin receptors perceive fast, non-genomic and slow, genomic auxin responses, respectively. Despite the fact that ABP1 mainly localizes to the endoplasmic reticulum (ER), until now it has been proposed to be active only in the extracellular matrix (reviewed in Sauer and Kleine-Vehn, 2011). Just recently, ABP1 function was also linked to genomic responses, modulating TIR1/AFB-dependent processes (Tromas et al., 2013). Intriguingly, the genomic effect of ABP1 appears to be at least partially independent of the endogenous auxin indole 3-acetic acid (IAA) (Paque et al., 2014). In this proposal my main research objective is to unravel the importance of the ER for genomic auxin responses. The PIN-LIKES (PILS) putative carriers for auxinic compounds also localize to the ER and determine the cellular sensitivity to auxin. PILS5 gain-of-function reduces canonical auxin signaling (Barbez et al., 2012) and phenocopies abp1 knock down lines (Barbez et al., 2012, Paque et al., 2014). Accordingly, a PILS-dependent substrate could be a negative regulator of ABP1 function in the ER. Based on our unpublished data, an IAA metabolite could play a role in ABP1-dependent processes in the ER, possibly providing feedback on the canonical nuclear IAA-signaling. I hypothesize that the genomic auxin response may be an integration of auxin- and auxin-metabolite-dependent nuclear and ER localized signaling, respectively. This proposed project aims to characterize a novel auxin-signaling paradigm in plants. We will employ state of the art interdisciplinary (biochemical, biophysical, computational modeling, molecular, and genetic) methods to assess the projected research. The identification of the proposed auxin conjugate-dependent signal could have far reaching plant developmental and biotechnological importance.
Project acronym BACEMO
Project Bacterial Cell Morphogenesis
Researcher (PI) Rut Carballido Lopez
Host Institution (HI) INSTITUT NATIONAL DE LA RECHERCHE AGRONOMIQUE
Summary In bacteria, the though external cell wall and the intracellular actin-like (MreB) cytoskeleton are major determinants of cell shape. The biosynthetic pathways and chemical composition of the cell wall, a three dimensional polymer network that is one of the most prominent targets for antibiotics, are well understood. However, despite decades of study, little is known about the complex cell wall ultrastructure and the molecular mechanisms that control cell wall morphogenesis in time and space. In rod-shaped bacteria, MreB homologues assemble into dynamic structures thought to control shape by serving as organizers for the movement and assembly of macromolecular machineries that effect sidewall elongation. However, the mechanistic details used by the MreB cytoskeleton to fulfil this role remain to be elucidated. Furthermore, development of high-resolution microscopy techniques has led to new breakthroughs this year, published by our lab and others, which are shaking the model developed over the last decade and re-questioning the MreB "actin cytoskeleton" designation. The aim of this project is to combine powerful genetic, biochemical, genomic and systems biology approaches available in the model bacterium Bacillus subtilis with modern high-resolution light microscopic techniques to study the dynamics and mechanistic details of the MreB cytoskeleton and of CW assembly. Parameters measured by the different approaches will be combined to quantitatively describe the features of bacterial cell morphogenesis.
In bacteria, the though external cell wall and the intracellular actin-like (MreB) cytoskeleton are major determinants of cell shape. The biosynthetic pathways and chemical composition of the cell wall, a three dimensional polymer network that is one of the most prominent targets for antibiotics, are well understood. However, despite decades of study, little is known about the complex cell wall ultrastructure and the molecular mechanisms that control cell wall morphogenesis in time and space. In rod-shaped bacteria, MreB homologues assemble into dynamic structures thought to control shape by serving as organizers for the movement and assembly of macromolecular machineries that effect sidewall elongation. However, the mechanistic details used by the MreB cytoskeleton to fulfil this role remain to be elucidated. Furthermore, development of high-resolution microscopy techniques has led to new breakthroughs this year, published by our lab and others, which are shaking the model developed over the last decade and re-questioning the MreB "actin cytoskeleton" designation. The aim of this project is to combine powerful genetic, biochemical, genomic and systems biology approaches available in the model bacterium Bacillus subtilis with modern high-resolution light microscopic techniques to study the dynamics and mechanistic details of the MreB cytoskeleton and of CW assembly. Parameters measured by the different approaches will be combined to quantitatively describe the features of bacterial cell morphogenesis.
Project acronym BACTERIAL SPORES
Project Investigating the Nature of Bacterial Spores
Researcher (PI) Sigal Ben-Yehuda
Summary When triggered by nutrient limitation, the Gram-positive bacterium Bacillus subtilis and its relatives enter a pathway of cellular differentiation culminating in the formation of a dormant cell type called a spore, the most resilient cell type known. Bacterial spores can survive for long periods of time and are able to endure extremes of heat, radiation and chemical assault. Remarkably, dormant spores can rapidly convert back to actively growing cells by a process called germination. Consequently, spore forming bacteria, including dangerous pathogens, (such as C. botulinum and B. anthracis) are highly resistant to antibacterial treatments and difficult to eradicate. Despite significant advances in our understanding of the process of spore formation, little is known about the nature of the mature spore. It is unrevealed how dormancy is maintained within the spore and how it is ceased, as the organization and the dynamics of the spore macromolecules remain obscure. The unusual biochemical and biophysical characteristics of the dormant spore make it a challenging biological system to investigate using conventional methods, and thus set the need to develop innovative approaches to study spore biology. We propose to explore the nature of spores by using B. subtilis as a primary experimental system. We intend to: (1) define the architecture of the spore chromosome, (2) track the complexity and fate of mRNA and protein molecules during sporulation, dormancy and germination, (3) revisit the basic notion of the spore dormancy (is it metabolically inert?), (4) compare the characteristics of bacilli spores from diverse ecophysiological groups, (5) investigate the features of spores belonging to distant bacterial genera, (6) generate an integrative database that categorizes the molecular features of spores. Our study will provide original insights and introduce novel concepts to the field of spore biology and may help devise innovative ways to combat spore forming pathogens.
When triggered by nutrient limitation, the Gram-positive bacterium Bacillus subtilis and its relatives enter a pathway of cellular differentiation culminating in the formation of a dormant cell type called a spore, the most resilient cell type known. Bacterial spores can survive for long periods of time and are able to endure extremes of heat, radiation and chemical assault. Remarkably, dormant spores can rapidly convert back to actively growing cells by a process called germination. Consequently, spore forming bacteria, including dangerous pathogens, (such as C. botulinum and B. anthracis) are highly resistant to antibacterial treatments and difficult to eradicate. Despite significant advances in our understanding of the process of spore formation, little is known about the nature of the mature spore. It is unrevealed how dormancy is maintained within the spore and how it is ceased, as the organization and the dynamics of the spore macromolecules remain obscure. The unusual biochemical and biophysical characteristics of the dormant spore make it a challenging biological system to investigate using conventional methods, and thus set the need to develop innovative approaches to study spore biology. We propose to explore the nature of spores by using B. subtilis as a primary experimental system. We intend to: (1) define the architecture of the spore chromosome, (2) track the complexity and fate of mRNA and protein molecules during sporulation, dormancy and germination, (3) revisit the basic notion of the spore dormancy (is it metabolically inert?), (4) compare the characteristics of bacilli spores from diverse ecophysiological groups, (5) investigate the features of spores belonging to distant bacterial genera, (6) generate an integrative database that categorizes the molecular features of spores. Our study will provide original insights and introduce novel concepts to the field of spore biology and may help devise innovative ways to combat spore forming pathogens.
Project acronym BCLYM
Project Molecular mechanisms of mature B cell lymphomagenesis
Researcher (PI) Almudena Ramiro
Host Institution (HI) CENTRO NACIONAL DE INVESTIGACIONESCARDIOVASCULARES CARLOS III (F.S.P.)
Summary Most of the lymphomas diagnosed in the western world are originated from mature B cells. The hallmark of these malignancies is the presence of recurrent chromosome translocations that usually involve the immunoglobulin loci and a proto-oncogene. As a result of the translocation event the proto-oncogene becomes deregulated under the influence of immunoglobulin cis sequences thus playing an important role in the etiology of the disease. Upon antigen encounter mature B cells engage in the germinal center reaction, a complex differentiation program of critical importance to the development of the secondary immune response. The germinal center reaction entails the somatic remodelling of immunoglobulin genes by the somatic hypermutation and class switch recombination reactions, both of which are triggered by Activation Induced Deaminase (AID). We have previously shown that AID also initiates lymphoma-associated c-myc/IgH chromosome translocations. In addition, the germinal center reaction involves a fine-tuned balance between intense B cell proliferation and program cell death. This environment seems to render B cells particularly vulnerable to malignant transformation. We aim at studying the molecular events responsible for B cell susceptibility to lymphomagenesis from two perspectives. First, we will address the role of AID in the generation of lymphomagenic lesions in the context of AID specificity and transcriptional activation. Second, we will approach the regulatory function of microRNAs of AID-dependent, germinal center events. The proposal aims at the molecular understanding of a process that lies in the interface of immune regulation and oncogenic transformation and therefore the results will have profound implications both to basic and clinical understanding of lymphomagenesis.
Most of the lymphomas diagnosed in the western world are originated from mature B cells. The hallmark of these malignancies is the presence of recurrent chromosome translocations that usually involve the immunoglobulin loci and a proto-oncogene. As a result of the translocation event the proto-oncogene becomes deregulated under the influence of immunoglobulin cis sequences thus playing an important role in the etiology of the disease. Upon antigen encounter mature B cells engage in the germinal center reaction, a complex differentiation program of critical importance to the development of the secondary immune response. The germinal center reaction entails the somatic remodelling of immunoglobulin genes by the somatic hypermutation and class switch recombination reactions, both of which are triggered by Activation Induced Deaminase (AID). We have previously shown that AID also initiates lymphoma-associated c-myc/IgH chromosome translocations. In addition, the germinal center reaction involves a fine-tuned balance between intense B cell proliferation and program cell death. This environment seems to render B cells particularly vulnerable to malignant transformation. We aim at studying the molecular events responsible for B cell susceptibility to lymphomagenesis from two perspectives. First, we will address the role of AID in the generation of lymphomagenic lesions in the context of AID specificity and transcriptional activation. Second, we will approach the regulatory function of microRNAs of AID-dependent, germinal center events. The proposal aims at the molecular understanding of a process that lies in the interface of immune regulation and oncogenic transformation and therefore the results will have profound implications both to basic and clinical understanding of lymphomagenesis.
Project acronym BRAINGAIN
Project NOVEL STRATEGIES FOR BRAIN REGENERATION
Researcher (PI) Andras Simon
Summary In contrast to mammals, newts possess exceptional capacities among vertebrates to rebuild complex structures, such as the brain. Our goal is to bridge the gap in the regenerative outcomes between newts and mammals. My group has made significant contributions towards this goal. We created a novel experimental system, which recapitulates central features of Parkinson's disease in newts, and provides a unique model for understanding regeneration in the adult midbrain. We showed an unexpected but key feature of the newt brain that it is akin to the mammalian brain in terms of the extent of homeostatic cell turn over, but distinct in terms of its injury response, showing the regenerative capacity of the adult vertebrate brain by activating neurogenesis in normally quiescent regions. Further we established a critical role for the neurotransmitter dopamine in controlling quiescence in the midbrain, thereby preventing neurogenesis during homeostasis and terminating neurogenesis once the correct number of neurons has been produced during regeneration. Here we aim to identify key molecular pathways that regulate adult neurogenesis, to define lineage relationships between neuronal stem and progenitor cells, and to identify essential differences between newts and mammals. We will combine pharmacological modulation of neurotransmitter signaling with extensive cellular fate mapping approaches, and molecular manipulations. Ultimately we will test hypotheses derived from newt studies with mammalian systems including newt/mouse cross species complementation approaches. We expect that our findings will provide new regenerative strategies, and reveal fundamental aspects of cell fate determination, tissue growth, and tissue maintenance in normal and pathological conditions.
In contrast to mammals, newts possess exceptional capacities among vertebrates to rebuild complex structures, such as the brain. Our goal is to bridge the gap in the regenerative outcomes between newts and mammals. My group has made significant contributions towards this goal. We created a novel experimental system, which recapitulates central features of Parkinson's disease in newts, and provides a unique model for understanding regeneration in the adult midbrain. We showed an unexpected but key feature of the newt brain that it is akin to the mammalian brain in terms of the extent of homeostatic cell turn over, but distinct in terms of its injury response, showing the regenerative capacity of the adult vertebrate brain by activating neurogenesis in normally quiescent regions. Further we established a critical role for the neurotransmitter dopamine in controlling quiescence in the midbrain, thereby preventing neurogenesis during homeostasis and terminating neurogenesis once the correct number of neurons has been produced during regeneration. Here we aim to identify key molecular pathways that regulate adult neurogenesis, to define lineage relationships between neuronal stem and progenitor cells, and to identify essential differences between newts and mammals. We will combine pharmacological modulation of neurotransmitter signaling with extensive cellular fate mapping approaches, and molecular manipulations. Ultimately we will test hypotheses derived from newt studies with mammalian systems including newt/mouse cross species complementation approaches. We expect that our findings will provide new regenerative strategies, and reveal fundamental aspects of cell fate determination, tissue growth, and tissue maintenance in normal and pathological conditions.
Project acronym CAPSEVO
Project Evolution of flower morphology: the selfing syndrome in Capsella
Researcher (PI) Michael Lenhard
Host Institution (HI) UNIVERSITAET POTSDAM
Summary The change from reproduction by outbreeding to selfing is one of the most frequent evolutionary transitions in plants. This transition is generally accompanied by changes in flower morphology and function, termed the selfing syndrome, including a reduction in flower size and a more closed flower structure. While the loss of self-incompatibility is relatively well understood, little is known about the molecular basis of the associated morphological changes and their evolutionary history. We will address these problems using the species pair Capsella grandiflora (the ancestral outbreeder) and C. rubella (the derived selfing species) as a genetically tractable model. We have established recombinant inbred lines from a cross of C. grandiflora x C. rubella and mapped quantitative trait loci affecting flower size and flower opening. Using this resource, the proposal will address four objectives. (1) We will isolate causal genes underlying the variation in flower size and opening, by combining genetic mapping with next-generation sequencing. (2) We will characterize the developmental and molecular functions of the isolated genes in Capsella and Arabidopsis. (3) We will dissect the molecular basis of the different allelic effects of the causal genes to determine which kinds of mutations have led to the morphological changes. (4) Based on population-genetic analyses of the isolated genes, the evolutionary history of the morphological changes will be retraced. Together, these strands of investigation will provide a detailed understanding of general processes underlying morphological evolution in plants.
The change from reproduction by outbreeding to selfing is one of the most frequent evolutionary transitions in plants. This transition is generally accompanied by changes in flower morphology and function, termed the selfing syndrome, including a reduction in flower size and a more closed flower structure. While the loss of self-incompatibility is relatively well understood, little is known about the molecular basis of the associated morphological changes and their evolutionary history. We will address these problems using the species pair Capsella grandiflora (the ancestral outbreeder) and C. rubella (the derived selfing species) as a genetically tractable model. We have established recombinant inbred lines from a cross of C. grandiflora x C. rubella and mapped quantitative trait loci affecting flower size and flower opening. Using this resource, the proposal will address four objectives. (1) We will isolate causal genes underlying the variation in flower size and opening, by combining genetic mapping with next-generation sequencing. (2) We will characterize the developmental and molecular functions of the isolated genes in Capsella and Arabidopsis. (3) We will dissect the molecular basis of the different allelic effects of the causal genes to determine which kinds of mutations have led to the morphological changes. (4) Based on population-genetic analyses of the isolated genes, the evolutionary history of the morphological changes will be retraced. Together, these strands of investigation will provide a detailed understanding of general processes underlying morphological evolution in plants.
Project acronym CBSCS
Project Physiology of the adult carotid body stem cell niche
Researcher (PI) Ricardo Pardal
Host Institution (HI) UNIVERSIDAD DE SEVILLA
Summary The discovery of adult neural stem cells (NSCs) has broaden our view of the physiological plasticity of the nervous system, and has opened new perspectives on the possibility of tissue regeneration and repair in the brain. NSCs reside in specialized niches in the adult mammalian nervous system, where they are exposed to specific paracrine signals regulating their behavior. These neural progenitors are generally in a quiescent state within their niche, and they activate their proliferation depending on tissue regenerative and growth needs. Understanding the mechanisms by which NSCs enter and exit the quiescent state is crucial for the comprehension of the physiology of the adult nervous system. In this project we will study the behavior of a specific subpopulation of adult neural stem cells recently described by our group in the carotid body (CB). This small organ constitutes the most important chemosensor of the peripheral nervous system and has neuronal glomus cells responsible for the chemosensing, and glia-like sustentacular cells which were thought to have just a supportive role. We recently described that these sustentacular cells are dormant stem cells able to activate their proliferation in response to a physiological stimulus like hypoxia, and to differentiate into new glomus cells necessary for the adaptation of the organ. Due to our precise experimental control of the activation and deactivation of the CB neurogenic niche, we believe the CB is an ideal model to study fundamental questions about adult neural stem cell physiology and the interaction with the niche. We propose to study the cellular and molecular mechanisms by which these carotid body stem cells enter and exit the quiescent state, which will help us understand the physiology of adult neurogenic niches. Likewise, understanding this neurogenic process will improve the efficacy of using glomus cells for cell therapy against neurological disease, and might help us understand some neural tumors.
The discovery of adult neural stem cells (NSCs) has broaden our view of the physiological plasticity of the nervous system, and has opened new perspectives on the possibility of tissue regeneration and repair in the brain. NSCs reside in specialized niches in the adult mammalian nervous system, where they are exposed to specific paracrine signals regulating their behavior. These neural progenitors are generally in a quiescent state within their niche, and they activate their proliferation depending on tissue regenerative and growth needs. Understanding the mechanisms by which NSCs enter and exit the quiescent state is crucial for the comprehension of the physiology of the adult nervous system. In this project we will study the behavior of a specific subpopulation of adult neural stem cells recently described by our group in the carotid body (CB). This small organ constitutes the most important chemosensor of the peripheral nervous system and has neuronal glomus cells responsible for the chemosensing, and glia-like sustentacular cells which were thought to have just a supportive role. We recently described that these sustentacular cells are dormant stem cells able to activate their proliferation in response to a physiological stimulus like hypoxia, and to differentiate into new glomus cells necessary for the adaptation of the organ. Due to our precise experimental control of the activation and deactivation of the CB neurogenic niche, we believe the CB is an ideal model to study fundamental questions about adult neural stem cell physiology and the interaction with the niche. We propose to study the cellular and molecular mechanisms by which these carotid body stem cells enter and exit the quiescent state, which will help us understand the physiology of adult neurogenic niches. Likewise, understanding this neurogenic process will improve the efficacy of using glomus cells for cell therapy against neurological disease, and might help us understand some neural tumors.
Project acronym CELL HORMONE
Project Bringing into focus the cellular dynamics of the plant growth hormone gibberellin
Researcher (PI) Alexander Morgan JONES
Summary During an organism's development it must integrate internal and external information. An example in plants, whose development stretches across their lifetime, is the coordination between environmental stimuli and endogenous cues on regulating the key hormone gibberellin (GA). The present challenge is to understand how these diverse signals influence GA levels and how GA signalling leads to diverse GA responses. This challenge is deepened by a fundamental problem in hormone research: the specific responses directed by a given hormone often depend on the cell-type, timing, and amount of hormone accumulation, but hormone concentrations are most often assessed at the organism or tissue level. Our approach, based on a novel optogenetic biosensor, GA Perception Sensor 1 (GPS1), brings the goal of high-resolution quantification of GA in vivo within reach. In plants expressing GPS1, we observe gradients of GA in elongating root and shoot tissues. We now aim to understand how a series of independently tunable enzymatic and transport activities combine to articulate the GA gradients that we observe. We further aim to discover the mechanisms by which endogenous and environmental signals regulate these GA enzymes and transporters. Finally, we aim to understand how one of these signals, light, regulates GA patterns to influence dynamic cell growth and organ behavior. Our overarching goal is a systems level understanding of the signal integration upstream and growth programming downstream of GA. The groundbreaking aspect of this proposal is our focus at the cellular level, and we are uniquely positioned to carry out our multidisciplinary aims involving biosensor engineering, innovative imaging, and multiscale modelling. We anticipate that the discoveries stemming from this project will provide the detailed understanding necessary to make strategic interventions into GA dynamic patterning in crop plants for specific improvements in growth, development, and environmental responses.
During an organism's development it must integrate internal and external information. An example in plants, whose development stretches across their lifetime, is the coordination between environmental stimuli and endogenous cues on regulating the key hormone gibberellin (GA). The present challenge is to understand how these diverse signals influence GA levels and how GA signalling leads to diverse GA responses. This challenge is deepened by a fundamental problem in hormone research: the specific responses directed by a given hormone often depend on the cell-type, timing, and amount of hormone accumulation, but hormone concentrations are most often assessed at the organism or tissue level. Our approach, based on a novel optogenetic biosensor, GA Perception Sensor 1 (GPS1), brings the goal of high-resolution quantification of GA in vivo within reach. In plants expressing GPS1, we observe gradients of GA in elongating root and shoot tissues. We now aim to understand how a series of independently tunable enzymatic and transport activities combine to articulate the GA gradients that we observe. We further aim to discover the mechanisms by which endogenous and environmental signals regulate these GA enzymes and transporters. Finally, we aim to understand how one of these signals, light, regulates GA patterns to influence dynamic cell growth and organ behavior. Our overarching goal is a systems level understanding of the signal integration upstream and growth programming downstream of GA. The groundbreaking aspect of this proposal is our focus at the cellular level, and we are uniquely positioned to carry out our multidisciplinary aims involving biosensor engineering, innovative imaging, and multiscale modelling. We anticipate that the discoveries stemming from this project will provide the detailed understanding necessary to make strategic interventions into GA dynamic patterning in crop plants for specific improvements in growth, development, and environmental responses.
Project acronym CELLONGATE
Project Unraveling the molecular network that drives cell growth in plants
Researcher (PI) Matyas FENDRYCH
Summary Plants differ strikingly from animals by the almost total absence of cell migration in their development. Plants build their bodies using a hydrostatic skeleton that consists of pressurized cells encased by a cell wall. Consequently, plant cells cannot migrate and must sculpture their bodies by orientation of cell division and precise regulation of cell growth. Cell growth depends on the balance between internal cell pressure – turgor, and strength of the cell wall. Cell growth is under a strict developmental control, which is exemplified in the Arabidopsis thaliana root tip, where massive cell elongation occurs in a defined spatio-temporal developmental window. Despite the immobility of their cells, plant organs move to optimize light and nutrient acquisition and to orient their bodies along the gravity vector. These movements depend on differential regulation of cell elongation across the organ, and on response to the phytohormone auxin. Even though the control of cell growth is in the epicenter of plant development, protein networks steering the developmental growth onset, coordination and termination remain elusive. Similarly, although auxin is the central regulator of growth, the molecular mechanism of its effect on root growth is unknown. In this project, I will establish a unique microscopy setup for high spatio-temporal resolution live-cell imaging equipped with a microfluidic lab-on-chip platform optimized for growing roots, to enable analysis and manipulation of root growth physiology. I will use developmental gradients in the root to discover genes that steer cellular growth, by correlating transcriptome profiles of individual cell types with the cell size. In parallel, I will exploit the auxin effect on root to unravel molecular mechanisms that control cell elongation. Finally, I am going to combine the live-cell imaging methodology with the gene discovery approaches to chart a dynamic spatio-temporal physiological map of a growing Arabidopsis root.
Plants differ strikingly from animals by the almost total absence of cell migration in their development. Plants build their bodies using a hydrostatic skeleton that consists of pressurized cells encased by a cell wall. Consequently, plant cells cannot migrate and must sculpture their bodies by orientation of cell division and precise regulation of cell growth. Cell growth depends on the balance between internal cell pressure – turgor, and strength of the cell wall. Cell growth is under a strict developmental control, which is exemplified in the Arabidopsis thaliana root tip, where massive cell elongation occurs in a defined spatio-temporal developmental window. Despite the immobility of their cells, plant organs move to optimize light and nutrient acquisition and to orient their bodies along the gravity vector. These movements depend on differential regulation of cell elongation across the organ, and on response to the phytohormone auxin. Even though the control of cell growth is in the epicenter of plant development, protein networks steering the developmental growth onset, coordination and termination remain elusive. Similarly, although auxin is the central regulator of growth, the molecular mechanism of its effect on root growth is unknown. In this project, I will establish a unique microscopy setup for high spatio-temporal resolution live-cell imaging equipped with a microfluidic lab-on-chip platform optimized for growing roots, to enable analysis and manipulation of root growth physiology. I will use developmental gradients in the root to discover genes that steer cellular growth, by correlating transcriptome profiles of individual cell types with the cell size. In parallel, I will exploit the auxin effect on root to unravel molecular mechanisms that control cell elongation. Finally, I am going to combine the live-cell imaging methodology with the gene discovery approaches to chart a dynamic spatio-temporal physiological map of a growing Arabidopsis root.
Project acronym CELLPATTERN
Project The Cellular Basis of Multicellular Pattern Formation
Researcher (PI) Dolf Weijers
Host Institution (HI) WAGENINGEN UNIVERSITY
Summary The formation of plant organs (leaves, roots, flowers) depends on the activity of stem cells (SC), located in stem cell niches (meristems) together with adjoining organizer cells (OC) that prevent SC differentiation. Despite their importance, SC and OC have been poorly described at molecular and cellular level and mechanisms for their coordinated specification are only partially understood. We study the specification of the very first SC and OC for the root in the early Arabidopsis embryo where cell divisions are almost invariant and, in the absence of cell motility, highly predictable. Previously we have established a central role for the transcription factor MONOPTEROS (MP) in OC specification and we have recently found that MP also controls SC specification. Hence, MP offers a unique entry point into studying the genomic and cellular reprogramming that underlies coordinated SC and OC specification. Our recent identification of MP target genes has shown that its function in SC specification is cell-autonomous, while MP-dependent OC specification involves a mobile transcription factor. In recent years we have developed a set of resources to systematically study embryonic root meristem initiation, and are now in a unique position to answer the following questions in this ERC project: 1. What transcriptional reprogramming underlies the first specification of SC and OC in the plant embryo? 2. What cellular changes follow from transcriptional reprogramming and mediate elongation and asymmetric division of SC and OC? 3. What is the mechanism of directional protein transport that ensures spatiotemporal coordination between SC and OC? The project will provide genome-wide insight in the cellular reprogramming underlying the coordinated formation of a multicellular structure. Finally, this work will shed light on mechanisms of stem cell and stem cell niche formation.
The formation of plant organs (leaves, roots, flowers) depends on the activity of stem cells (SC), located in stem cell niches (meristems) together with adjoining organizer cells (OC) that prevent SC differentiation. Despite their importance, SC and OC have been poorly described at molecular and cellular level and mechanisms for their coordinated specification are only partially understood. We study the specification of the very first SC and OC for the root in the early Arabidopsis embryo where cell divisions are almost invariant and, in the absence of cell motility, highly predictable. Previously we have established a central role for the transcription factor MONOPTEROS (MP) in OC specification and we have recently found that MP also controls SC specification. Hence, MP offers a unique entry point into studying the genomic and cellular reprogramming that underlies coordinated SC and OC specification. Our recent identification of MP target genes has shown that its function in SC specification is cell-autonomous, while MP-dependent OC specification involves a mobile transcription factor. In recent years we have developed a set of resources to systematically study embryonic root meristem initiation, and are now in a unique position to answer the following questions in this ERC project: 1. What transcriptional reprogramming underlies the first specification of SC and OC in the plant embryo? 2. What cellular changes follow from transcriptional reprogramming and mediate elongation and asymmetric division of SC and OC? 3. What is the mechanism of directional protein transport that ensures spatiotemporal coordination between SC and OC? The project will provide genome-wide insight in the cellular reprogramming underlying the coordinated formation of a multicellular structure. Finally, this work will shed light on mechanisms of stem cell and stem cell niche formation.
Project acronym CELLREPROGRAMMING
Project Uncovering the Mechanisms of Epigenetic Reprogramming of Pluripotent and Somatic Cell States
Summary The generation of animals by nuclear transfer demonstrated that the epigenetic state of somatic cells could be reset to an embryonic state, capable of directing the development of a new organism. The nuclear cloning technology is of interest for transplantation medicine, but any application is hampered by the inefficiency and ethical problems. A breakthrough solving these issues has been the in vitro derivation of reprogrammed Induced Pluripotent Stem "iPS" cells by the ectopic expression of defined transcription factors in somatic cells. iPS cells recapitulate all defining features of embryo-derived pluripotent stem cells, including the ability to differentiate into all somatic cell types. Further, recent publications have demonstrated the ability to directly trans-differentiate somatic cell types by ectopic expression of lineage specification factors. Thus, it is becoming increasingly clear that an ultimate goal in the stem cell field is to enable scientists to have the power to safely manipulate somatic cells by "reprogramming" their behavior at will. However, to frame this challenge, we must understand the basic mechanisms underlying the generation of reprogrammed cells in parallel to designing strategies for their medical application and their use in human disease specific research. In this ERC Starting Grant proposal, I describe comprehensive lines of experimentation that I plan to conduct in my new lab scheduled to open in April 2011 at the Weizmann Institute of Science. We will utilize exacting transgenic mammalian models and high throughput sequencing and genomic screening tools for in depth characterization of the molecular "rules" of rewiring the epigenome of somatic and pluripotent cell states. The proposed research endeavors will not only contribute to the development of safer strategies for cell reprogramming, but will also help decipher how diverse gene expression programs lead to cellular specification during normal development.
The generation of animals by nuclear transfer demonstrated that the epigenetic state of somatic cells could be reset to an embryonic state, capable of directing the development of a new organism. The nuclear cloning technology is of interest for transplantation medicine, but any application is hampered by the inefficiency and ethical problems. A breakthrough solving these issues has been the in vitro derivation of reprogrammed Induced Pluripotent Stem "iPS" cells by the ectopic expression of defined transcription factors in somatic cells. iPS cells recapitulate all defining features of embryo-derived pluripotent stem cells, including the ability to differentiate into all somatic cell types. Further, recent publications have demonstrated the ability to directly trans-differentiate somatic cell types by ectopic expression of lineage specification factors. Thus, it is becoming increasingly clear that an ultimate goal in the stem cell field is to enable scientists to have the power to safely manipulate somatic cells by "reprogramming" their behavior at will. However, to frame this challenge, we must understand the basic mechanisms underlying the generation of reprogrammed cells in parallel to designing strategies for their medical application and their use in human disease specific research. In this ERC Starting Grant proposal, I describe comprehensive lines of experimentation that I plan to conduct in my new lab scheduled to open in April 2011 at the Weizmann Institute of Science. We will utilize exacting transgenic mammalian models and high throughput sequencing and genomic screening tools for in depth characterization of the molecular "rules" of rewiring the epigenome of somatic and pluripotent cell states. The proposed research endeavors will not only contribute to the development of safer strategies for cell reprogramming, but will also help decipher how diverse gene expression programs lead to cellular specification during normal development.
Project acronym CENTRIOLSTRUCTNUMBER
Project Control of Centriole Structure And Number
Researcher (PI) Monica Bettencourt Carvalho Dias
Host Institution (HI) FUNDACAO CALOUSTE GULBENKIAN
Summary Centrioles are essential for the formation of several microtubule organizing structures including cilia, flagella and centrosomes. These structures are involved in a variety of functions, from cell motility to division. Centrosome defects are seen in many cancers, while abnormalities in cilia and flagella can lead to a variety of human diseases, such as polycystic kidney disease. The molecular mechanisms regulating centriole biogenesis have only recently started to be unravelled, opening new ways to answer a wide range of questions that have fascinated biologists for more than a century. In this grant we are asking two fundamental questions that are central to human disease: how is centriole structure and number established and regulated in the eukaryotic cell? To address these questions we propose to identify new molecular players, and to test the role of these and known players in the context of specific mechanistic hypothesis, using in vitro and in vivo models. We propose to develop novel assays for centriole structure and regulation in order to address mechanistic problems not accessible with today s assays. In our search for novel components we will use a multidisciplinary approach combining bioinformatics with high throughput screening. The use of in vitro systems will permit the quantitative dissection of molecular mechanisms, while the study of those mechanisms in Drosophila will allow us to understand them at the whole organism level. Furthermore, this analysis, together with studies in human tissue culture cells, will allow us to understand the consequences of misregulation of these fundamental centriole properties for human disease, such as ciliopathies and cancer. My group is already collaborating with medical doctors in the study of centriole aberrations in human disease (cancer and ciliopathies), which will be invaluable to bringing the results of this study to the translational level.
Centrioles are essential for the formation of several microtubule organizing structures including cilia, flagella and centrosomes. These structures are involved in a variety of functions, from cell motility to division. Centrosome defects are seen in many cancers, while abnormalities in cilia and flagella can lead to a variety of human diseases, such as polycystic kidney disease. The molecular mechanisms regulating centriole biogenesis have only recently started to be unravelled, opening new ways to answer a wide range of questions that have fascinated biologists for more than a century. In this grant we are asking two fundamental questions that are central to human disease: how is centriole structure and number established and regulated in the eukaryotic cell? To address these questions we propose to identify new molecular players, and to test the role of these and known players in the context of specific mechanistic hypothesis, using in vitro and in vivo models. We propose to develop novel assays for centriole structure and regulation in order to address mechanistic problems not accessible with today s assays. In our search for novel components we will use a multidisciplinary approach combining bioinformatics with high throughput screening. The use of in vitro systems will permit the quantitative dissection of molecular mechanisms, while the study of those mechanisms in Drosophila will allow us to understand them at the whole organism level. Furthermore, this analysis, together with studies in human tissue culture cells, will allow us to understand the consequences of misregulation of these fundamental centriole properties for human disease, such as ciliopathies and cancer. My group is already collaborating with medical doctors in the study of centriole aberrations in human disease (cancer and ciliopathies), which will be invaluable to bringing the results of this study to the translational level.
Project acronym CENTROSTEMCANCER
Project Investigating the link between centrosomes, stem cells and cancer
Researcher (PI) Renata Homem De Gouveia Xavier De Basto
Host Institution (HI) INSTITUT CURIE
Summary Centrosomes are cytoplasmic organelles found in most animal cells with important roles in polarity establishment and maintenance. Theodor Boveri s pioneering work first suggested that extra-centrosomes could contribute to genetic instability and consequently to tumourigenesis. Although many human tumours do exhibit centrosome amplification (extra centrosomes) or centrosome abnormalities, the exact contribution of centrosomes to tumour initiation in vertebrate organisms remains to be determined. I have recently showed that Drosophila flies carrying extra-centrosomes, following the over-expression of the centriole replication kinase Sak, did not exhibit chromosome segregation errors and were able to maintain a stable diploid genome over many generations. Surprisingly, however, neural stem cells fail frequently to align the mitotic spindle with their polarity axis during asymmetric division. Moreover, I have found that centrosome amplification is permissive to tumour formation in flies. So far, however, we do not know the molecular mechanisms that allow transformation when extra centrosomes are present and elucidating these mechanisms is the aim of the work presented in this proposal. Here, I describe a series of complementary approaches that will help us to decipher the link between centrosomes, stem cells and tumour biology. In addition, I wish to pursue the original observations made in Drosophila and investigate the consequences of centrosome amplification in mammals.
Centrosomes are cytoplasmic organelles found in most animal cells with important roles in polarity establishment and maintenance. Theodor Boveri s pioneering work first suggested that extra-centrosomes could contribute to genetic instability and consequently to tumourigenesis. Although many human tumours do exhibit centrosome amplification (extra centrosomes) or centrosome abnormalities, the exact contribution of centrosomes to tumour initiation in vertebrate organisms remains to be determined. I have recently showed that Drosophila flies carrying extra-centrosomes, following the over-expression of the centriole replication kinase Sak, did not exhibit chromosome segregation errors and were able to maintain a stable diploid genome over many generations. Surprisingly, however, neural stem cells fail frequently to align the mitotic spindle with their polarity axis during asymmetric division. Moreover, I have found that centrosome amplification is permissive to tumour formation in flies. So far, however, we do not know the molecular mechanisms that allow transformation when extra centrosomes are present and elucidating these mechanisms is the aim of the work presented in this proposal. Here, I describe a series of complementary approaches that will help us to decipher the link between centrosomes, stem cells and tumour biology. In addition, I wish to pursue the original observations made in Drosophila and investigate the consequences of centrosome amplification in mammals.
Project acronym CentSatRegFunc
Project Dissecting the function and regulation of centriolar satellites: key regulators of the centrosome/cilium complex
Researcher (PI) Elif Nur Firat Karalar
Host Institution (HI) KOC UNIVERSITY
Summary Centrosomes are the main microtubule-organizing centers of animal cells. They influence the morphology of the microtubule cytoskeleton and function as the base of primary cilium, a nexus for important signaling pathways. Structural and functional defects in centrosome/cilium complex cause a variety of human diseases including cancer, ciliopathies and microcephaly. To understand the relationship between human diseases and centrosome/cilium abnormalities, it is essential to elucidate the biogenesis of centrosome/cilium complex and the control mechanisms that regulate their structure and function. To tackle these fundamental problems, we will dissect the function and regulation of centriolar satellites, the array of granules that localize around the centrosome/cilium complex in mammalian cells. Only recently interest in the satellites has grown because mutations affecting satellite components were shown to cause ciliopathies, microcephaly and schizophrenia. Remarkably, many centrosome/cilium proteins localize to these structures and we lack understanding of when, why and how these proteins localize to satellites. The central hypothesis of this grant is that satellites ensure proper centrosome/cilium complex structure and function by acting as transit paths for modification, assembly, storage, stability and trafficking of centrosome/cilium proteins. In Aim 1, we will identify the nature of regulatory and molecular relationship between satellites and the centrosome/cilium complex. In Aim 2, we will elucidate the role of satellites in proteostasis of centrosome/cilium proteins. In Aim 3, we will investigate the functional significance of satellite-localization of centrosome/cilium proteins during processes that go awry in human disease. Using a multidisciplinary approach, the proposed research will expand our knowledge of the spatiotemporal regulation of the centrosome/cilium complex and provide new insights into pathogenesis of ciliopathies and primary microcephaly.
Centrosomes are the main microtubule-organizing centers of animal cells. They influence the morphology of the microtubule cytoskeleton and function as the base of primary cilium, a nexus for important signaling pathways. Structural and functional defects in centrosome/cilium complex cause a variety of human diseases including cancer, ciliopathies and microcephaly. To understand the relationship between human diseases and centrosome/cilium abnormalities, it is essential to elucidate the biogenesis of centrosome/cilium complex and the control mechanisms that regulate their structure and function. To tackle these fundamental problems, we will dissect the function and regulation of centriolar satellites, the array of granules that localize around the centrosome/cilium complex in mammalian cells. Only recently interest in the satellites has grown because mutations affecting satellite components were shown to cause ciliopathies, microcephaly and schizophrenia. Remarkably, many centrosome/cilium proteins localize to these structures and we lack understanding of when, why and how these proteins localize to satellites. The central hypothesis of this grant is that satellites ensure proper centrosome/cilium complex structure and function by acting as transit paths for modification, assembly, storage, stability and trafficking of centrosome/cilium proteins. In Aim 1, we will identify the nature of regulatory and molecular relationship between satellites and the centrosome/cilium complex. In Aim 2, we will elucidate the role of satellites in proteostasis of centrosome/cilium proteins. In Aim 3, we will investigate the functional significance of satellite-localization of centrosome/cilium proteins during processes that go awry in human disease. Using a multidisciplinary approach, the proposed research will expand our knowledge of the spatiotemporal regulation of the centrosome/cilium complex and provide new insights into pathogenesis of ciliopathies and primary microcephaly.


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Why Islamic State targets airports: A global stage and variety of nationalities
World Devparna Acharya Jul 04, 2016 15:39:53 IST
An alert from the SITE Intelligence Group on Friday warned that a "pro-Islamic State" account had issued threats against aircraft "flying from Heathrow to the US" this weekend, which coincides with the American Independence Day holiday, The Times reported. The alert warned of a potentially deadly device being placed at Heathrow, Los Angeles airport or John F Kennedy in New York.
Pro-IS Twitter Account Warns of Attacks in Heathrow, LAX, and JFK Airports on July 4 Weekend https://t.co/nResxCNiwL
— SITE Intel Group (@siteintelgroup) July 1, 2016
British transport minister Tariq Ahmad called for the public to be cautious.
"We all need to be vigilant to the global threat of terrorism — in the UK, we keep all aspects of aviation security under constant review and work closely with our international partners to mitigate risks," Ahmad said, adding, "We will continue to take all steps necessary to keep the public safe, but for security reasons we do not comment in detail on specific measures or operational matters." On Sunday, top intelligence chief of Germany warned that after Istanbul, the Islamic State may attack German airports, which are likely to be teeming with people from different nationalities.
With the blasts at Istanbul's international airport on its heel, the news alert came in at a time when worldwide panic was setting in regarding terrorist attacks. On Friday night, when the tweet surfaced, hostages were taken by Islamic terrorists in an upscale café in Dhaka. The siege carried on for almost 10 hours and killed at least 20.
A view of the departure terminal of Istanbul's Ataturk Airport. Reuters
There is a price to be paid for fighting terrorism — a price the people of too many nations, including our own, have paid in recent days. Especially after the brazen suicide bombing in Istanbul's Ataturk airport and the recent threat by the 'supporters' of Islamic State brings us to the inevitable question: Are airports an easy target for terrorists to wage a war?
While the investigation is in its early stages, it's clear that the attack on Ataturk airport was meticulously planned and executed. The assailants arrived in a taxi and went straight for one of the busiest areas of the airport. They also managed to overcome heightened security put in place at Ataturk after the March assault on Brussels airport.
Come to think of it, reasons as to how terrorist groups zero-in on a particular target and carry out a successful attack are many, but planning an attack in an airport — especially a vast international airport like the one in Istanbul (which is among the most secure in the world, likely second only to Tel Aviv in its number of checkpoints), is something worth looking into.
This particular act of terrorism was indeed exceedingly well-timed — intended no doubt to make an already jittery travelling public even more nervous at the height of the summer tourist season. But the attack also comes during a week in which Turkish president Recep Tayyip Erdogan has made two significant moves to restore fractured relationships with Israel and with Russia. Just on Sunday, Ankara reached an agreement to resume full diplomatic relations with Israel after six years of estrangement. On Monday, Erdogan issued a letter apologising to Russia for downing a jet near its border with Syria.
Bombing an airport, where security is probably the maximum, is making a statement as far as terrorists are concerned. It is about making a spectacle and send a message to world leaders as to how far they can reach. US Secretary of State John Kerry recently was quoted as saying that the Islamic State is targeting airports because they are desperate.
"It has been more than one year since Daesh (Islamic State) has actually launched a full scale military offensive, and that's because our coalition is moving relentlessly on every front. Now, yes, you can bomb an airport, you can blow yourself up. That's the tragedy. Daesh and others like it know that we have to get it right 24/7/365. They have to get it right for ten minutes or one hour, so it's a very different scale. And if you're desperate and if you know you are losing, and you know you want to give up your life, then obviously you can do some harm."
Harm was done. Forty-one were killed and hundreds were injured, not to mention the panic it sends every other nation into. Terrorists targeted the 11th busiest airport in the world and the 3rd busiest in Europe after London's Heathrow and Paris' Charles de Gaulle, in a country that borders the Islamic State-stronghold Syria, and it's not hard to see why Ataturk posed an attractive soft target.
According to this report on CNN, travellers at the Istanbul airport are required to undergo double security screenings. One before they enter the international terminal building, and a second time after they go through passport control. It also posts a vehicle checkpoint about 500 metres from the entrance of the international terminal — although only suspicious vehicles are stopped and checked, the report added. According to experts, who specialise in West Asian security issues, there is no foolproof method of preventing similar airport attacks. Issues like weaker security at arrivals compared to that of the departure wing and response of the security staff at airports, help in making these deadly attacks possible.
One of the reasons for this is that while tarmac and terminal security have been intensified, access to terminals and satellite buildings is still too easy across the world.
Image source: Reuters
But, as mentioned earlier, when such attacks are actually made possible, it is a matter of pride for the perpetrators. An airport is a location where you find people of all the nationalities, entering and leaving the country. All at one place. It is an ideal spot for terrorists to make a global statement and where their target base can potentially be internationally varied.
In a study done in 2004 by RAND, one fact that consistently emerged from the analysis: it is not the size of the bomb that matters most; it is where it is detonated. Usually terrorists, serious about making a strong impact, involve placement of a bomb in close proximity to a vulnerable crowd of people.
Counter-terrorism expert Greg Barton was quoted by the ABC news who said that airports were the softest target by militants. In 2016 March, 32 were killed and more than 300 were injured in three bombings in Belgium, including two at Brussels Airport. According to Barton, there is no easy solution to this. Airports cannot be shut down in the face of a suspected attack and in the case of Istambul airport, it has over 16 million people going through every year.
"We will have to look at the way we manage security in airports and in public places generally. If you had people intent on blowing themselves up in the course of this attack, there's absolutely nothing you can do to stop that.
However, analysts also said that regardless of any boosted security measures, airports are among the "most vulnerable" soft targets and likely always will be. Airports are extremely busy, security checkpoints and screening areas create masses of travellers standing and waiting to be processed making it equally hard for any security agency of coming up with an effective way of avoiding it. Moving the target (passengers) also does not yield any helpful results.
Gresham, Smith and Partners recently designed a screening area at Norfolk International Airport in Virginia with one major concern in mind: Flexibility, so it can adapt to changing security threats. From box-cutters to explosives to automatic weapons, the dangers for airport security evolve. So the firm created a large, open space without support columns that can be easily reconfigured to bring in the next generation of screening machines. "We don't know what's coming next so we design for that," said Wilson Rayfield, executive vice president in charge of aviation at the architecture, design and consulting firm.
In the face of airport threats, designers are asked to come to the frontline of the security challenge and achieve the nearly impossible: improve security without slowing down travelers. Sometimes, art and function coincide. Open spaces and high ceilings can reduce the impact of a concussive blast. Other times, designers are working to reduce congestion in non-secure areas and create more offsite checkpoints. They seek to channel passengers in ways that take advantage of high-tech sensors, cameras and facial recognition software that may help police stop assailants before they kill.
"Aviation has a lot to learn from Las Vegas casinos," said Rayfield, referring to surveillance cameras and crowd control methods that he said allow three-fourths of visitors to be identified.
Ofer Lefler, a spokesman for Israel Airports Authority, said security was "100 percent" a consideration required of architects who designed Ben Gurion Airport's main terminal, though he declined to discuss specifics. The terminal, completed in 2004, is grand with high ceilings and an abundance of marble and Jerusalem stone. A magnificent sun roof, water fountain and atrium give way to corridors leading to the gates like spokes on a wheel.
Beyond aesthetics, the design has a function, according to one Israeli aviation security consultant who spoke to Reuters on condition of anonymity as he was unauthorised to discuss measures at Ben Gurion Airport. Wide-open sight lines give security agents a clear view so that "potential terrorists can be tracked by guards, whether in person or through the closed-circuit TV system, from the moment they are arrive." In places under constant threat, like Baghdad and Kabul, security checkpoints begin miles from the terminal and include myriad scans, checks and bomb-sniffing dogs.
From the parking area to the terminal, there are several access points with sliding glass doors made from a blast-proof material that would help limit casualties from shrapnel, the consultant said. Surveillance at Ben Gurion begins well before anyone reaches the parking area or terminal. Cars are stopped at a checkpoint, watched over by heavily armed guards and cameras that read license plates. People deemed suspicious are pulled over for further questioning and possibly searches. Largely surreptitious monitoring continues all the way to the terminal.
But, experts say, such measures may be impractical at busier airports. Ben Gurion handled fewer than 16 million international passengers in 2015, compared to 75 million at London Heathrow. The threat of terrorism at airports is so persistent and ever-evolving that security measures must also adapt to reduce the likelihood of a high consequence event. Bombers have continued to identify different targets of opportunity within the airport environment as seen most recently in Brussels, but also in Karachi, Beijing, Moscow and Madrid over the past decade.
With inputs from Reuters
Updated Date: Jul 04, 2016 15:39:53 IST
Tags : Ataturk International Airport, ConnectTheDots, Heathrow Airport, India, Islamic-State, Istanbul, John F Kennedy Airport, Tel Aviv Airport, USA, World
'Drunk' Irish woman, who went on expletive-laden rant at Air India crew, found dead in London home; Sussex Police rules out foul play
In chilling warning to LeT, HM in Kashmir, Islamic State affiliate warns it 'will feel no weakness in slaying these enemies of Islam'
Yazidi women rescued from Islamic State captors face dilemma of leaving 'Daesh' children or being shunned by community in Iraq's Sinjar
Body of alleged 'stowaway' falls from Kenya Airways plane lands in London garden; probe underway
1Why Islamic State targets airports: A global stage and variety of nationalities

redpajama_set_name: RedPajamaCommonCrawl


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Sick and Tired of the Manufactured Crisis A GOVERNMENT FOR AND BY THE SHEEPLE
Tearing Apart our Social Fabric, Thread by Thread
ppjg families, HEALTH, The PPJ Gazette coronavirus, COVID, Covid-19, elderly deaths, families, HEALTH, Kaiser Family Foundation, Marilyn M. Singleton, MD JD, Michigan governor, politicians pay cuts, The PPJ Gazette, underlying medical conditions Leave a comment
By Marilyn M. Singleton, MD, JD
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It seems like some folks have used the ghost of Ernesto "Ché" Guevara as their guide through the COVID-19 epidemic in the United States. "To send men to the firing squad [job loss, suicide, substance abuse], judicial [scientific] proof is unnecessary… This is a revolution! And a revolutionary must become a cold killing machine motivated by pure hate [of President Trump]."
Apparently, Ché was on to something. Forty-three percent of U.S. adults and 52 percent of low income adults say they or someone in their household has lost a job or taken a pay cut due to the outbreak. How can any American stuck at home not be disgusted by politicians who are still collecting their full paychecks while the middle class and working poor descend into an abyss. To be fair, Michigan's governor announced that she would take a 10 percent pay cut in solidarity with the people who had a 100 percent pay cut. Nonetheless, many have unquestionably accepted the government's oft times contradictory mandates.
A Kaiser Family Foundation poll found that despite the disruption of their lives 80 percent of Americans say strict shelter-in-place measures are worth it to protect people. This blind acquiescence is evidenced by unthinking people wearing masks while driving alone in their cars with closed windows. And perversely, designer face masks are proliferating. Wearing a mask is a sometimes necessary, serious, unpleasant thing to do. It is not fun. It is not cute.
It is not only the evil Chinese Communists that caused panic about the coronavirus in the U.S. via text messages falsely saying President Trump was imminently going to lock down the entire country. The media are complicit in the hysteria. Again, Ché Guevara had it right: "Foreign reporters—preferably American—were much more valuable to us at that time than any military victory. Much more valuable than recruits for our guerrilla force, were American media recruits to export our propaganda." Media hacks are collecting their paychecks to scare our bodily fluids out of us with day in, day out, COVID, COVID, COVID. We're all going to die! (Of course, they don't discuss the 1,300 people a day who die of complications of high blood pressure.)
COVID-19's angel of death spares most people: 80 percent of COVID-19 deaths occurred among persons aged 60 years and over; about 25 percent of all deaths were sick and elderly residents of long term care facilities; nearly 90 percent of persons hospitalized have one or more underlying medical conditions. Oddly, the CDC is boosting the official death toll by including not only people who died due to COVID-19 but those who died of other causes and had the virus that causes COVID-19 in their system.
Antibody testing studies in Santa Clara and Los Angeles (California) and Miami Dade (Florida) counties revealed that infection with the virus that causes COVID-19 is more widespread than indicated by the number of confirmed cases. The mortality is likely 0.1 percent (like the seasonal flu). Many researchers concluded that the data indicated that the virus is widely circulating, we will get infected but likely not die, and the population may develop immunity. The tyrants-in-waiting use the data to mandate lock downs until every last vestige of COVID-19 has disappeared from the face of the earth or a vaccine materializes—which may be never.
Americans have been terrified into accepting tyranny and poverty, softened with assurances that the current state of affairs is "normal." The airwaves' constant blare of "we're all in this together" does not make it so. Some people remain more equal than others. Just ask Nancy Pelosi with her $24,000 refrigerator filled with $13 a pint ice cream that she "gets by" on. Or professional athletes who received their elective surgeries while the rest of the peasants were denied their cancer treatment or the treatment of a painful joint. Or the TV star who complained about being "in jail" in her $45 million home.
Meanwhile, liquor sales are up 55 percent, calls to suicide hotlines and suicides have dramatically increased, domestic violence is rising, and anti-anxiety medication prescriptions are up 34 percent since the virus consumed our lives. We are on the path to becoming fat, depressed, drunken wife-beaters. We are told to ignore statistics because every death is tragic. If the lockdowns persist, every life is a tragedy in the making.
This epidemic has become the opportunity to change the U.S. from a free, energetic, resourceful individualistic society to an authoritarian, collectivist society of broken souls addicted to government largesse. To wit, a Los Angeles version of wealth redistribution: People can't work so they can't pay their rent. The mom and pop landlords can't pay their property taxes. The city then buys the "distressed" properties from desperate landlords at cut-rate prices and turns them over to the homeless. Ché would be proud.
Bio: Dr. Singleton is a board-certified anesthesiologist. She is the immediate past President of the Association of American Physicians and Surgeons (AAPS). She graduated from Stanford and earned her MD at UCSF Medical School. Dr. Singleton completed 2 years of Surgery residency at UCSF, then her Anesthesia residency at Harvard's Beth Israel Hospital. While still working in the operating room, she attended UC Berkeley Law School, focusing on constitutional law and administrative law. She interned at the National Health Law Project and practiced insurance and health law. She teaches classes in the recognition of elder abuse and constitutional law for non-lawyers. She lives in Oakland, Ca.


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Focussing on the vertebrate
fauna of SE Asia
SE Asia fauna ...
Carnivorans
Large Mammals
Lizards & Crocodilians
FFrogs & other calls
SE Asia Vertebrate Records (SEAVR) 2019
New Guinea fauna ...
Search this site ...
Recently added ...
Wildlife Crime Hotline for Peninsular Malaysia
Links :
Primates Of Johor Brochure
Context Institute
Herpetological Soc. Singapore
HabitatID
Primatewatching
Orang Utan Appeal (UK)
Wallace Online
MYCAT
Nature Society (Singapore)
Text and photos by Nick Baker, unless otherwise credited.
Copyright © Ecology Asia 2019
> Kuala Selangor < back
Kuala Selangor Nature Park
... of milky stork, leaf monkeys and fireflies
View from the Main Hide across the brackish lakes
Kuala Selangor lies just 75 kilometres north-west of Kuala Lumpur, near the mouth of the Selangor River. In addition to it's small-town charms there are many nearby attractions to suit the nature lover, including the Kuala Selangor Nature Park and the colonies of fireflies living along tributaries of the Selangor River.
The Kuala Selangor Nature Park is managed by the Malaysian Nature Society, under a co-operative arrangement with the Selangor State Government. This coastal area was once dominated by mangroves, but since the construction of coastal bunds it now comprises a strip of remnant mangrove forests, mainly Avicennia sp., behind which are brackish lakes and ponds and a large area of secondary forest. In many respects the ecological history of the area is similar to Singapore's own Sungei Buloh Wetland Reserve. Thus, familiar vertebrates such as mud skippers, mud lobsters and the Malayan Water Monitor can be seen from the bund and the mangrove boardwalks.
The endangered Milky Stork Mycteria cinerea - a captive breeding program is aimed at preventing their extinction.
During the migration season, between September and March, the lakes and ponds teem with shorebirds such as plovers, Greenshank, Redshank and various species of sandpiper travelling the migration route of the East-Asian Flyway. These very same birds may well stop at Sungei Buloh for a final meal of worms and molluscs before heading south to winter in the warmth of Australia.
One bird deserves a special mention – the Milky Stork. In conjunction with Zoo Negara, an active breeding program of this globally endangered species - less than 100 specimens are thought to exist - is being conducted in a specially constructed aviary next to the main lake. Visitors are not allowed inside the aviary, however the birds can clearly be seen nesting in the enclosure. It is hoped, upon release, these captive-bred storks will choose to make Kuala Selangor their permanent home.
Kuala Selangor Nature Park has over 200-hectares of immature secondary forest. Easy walks are possible along the well worn trails; where the sun streams through breaks in the canopy keep an eye open for lizards and skinks warming themselves, especially the Common Sun Skink. Familiar forest birds such as Woodpeckers, Bulbuls, Sunbirds and Malkohas can also be seen.
The Silvered Leaf Monkey or Silvered Langur Presbytis cristata - common in the secondary forest of Kuala Selangor Nature Park
Silvered Leaf Monkeys
One highly vocal species the visitor is likely to hear, if not see, in the secondary forest is the Silvered Leaf Monkey, also called the Silvered Langur. Troops of these large, grey-black, long-tailed monkeys can be heard crashing through the trees, scaring away the more timid bird species. In the same manner as trendy young people, the baby langurs appear to have died their fur a startling bright orange colour; one wonders how the adult monkeys cope with being the parents of such outrageous offspring ! In the early morning the langurs raid the gardens of people living at the base of Bukit Melawati, at the northern edge of the park, searching for ripened fruits.
Well marked trails wind their way through the secondary forest
The mischievous Long-tailed Macaque, which is common all over Malaysia and Singapore, appears to be less common here. Presumably, there is competition for food and territory between the macaques and the langurs; the smaller macaques must content themselves with living on the park's boundaries.
Kelip-kelip
Away from the Nature Park, branching off the Selangor River are a series of narrower tributaries lined with healthy stands of Beremban or Sonneratia Caseolaris. This tall mangrove species is characterised by simple symmetric leaves, and cone-like aerial roots. During the day this tree species might seem unremarkable, however as dusk falls beautiful flowers open up, each with an array of up to 100 reddish-white stamens. In these quiet, ulu backwaters survive Pteroptyx tener, the firefly species which makes S. Caseolaris their home.
Pteroptyx sp. - the beetle species known as "fireflies", or "kelip-kelip" in Malay.
At twilight, the kingfishers and bee-eaters give way to nightjars and owls, and the fireflies, or "kelip-kelip" as they are locally known, put on a dazzling light show. Thousands of these insects commence their fluorescent display, flashing on and off two or three times a second, some stationary, others circling around. In some trees a whole colony of fireflies will flash in absolute unison, hour after hour, in the manner of the lights on a Christmas tree. Male and female fireflies are both able to flash, but only the males flash in unison; the synchronised display serves to attract a mate, but how does the female choose her mate amongst the thousands of "switched-on" males !
Underside view of Pteroptyx sp. showing the light-emitting abdominal segments
The dense colonies of fireflies were, in olden times, used as a navigation aid in some rivers in Malaysia. As you view this wonderful sight, lulled by the gently rocking boat and the quiet putter of the engine, you fancy that thousands of years ago when the world was less polluted, many rivers supported these wondrous insects. Imagine the awe of primitive humans as they migrated down the Malay Peninsula thousands of years ago and their eyes saw these flashing lights for the first time.
Now the fireflies can be easily viewed by modern humans at Kampung Kuantan and Kampung Bukit Belimbing, both a short taxi ride from Kuala Selangor town. At both locations tourist facilities have grown in the last few years, so there are new jetties, reliable boatmen and restaurant and toilet facilities. If you think that the commercialisation of firefly viewing could in some way be a threat to their survival, think again. Consider that a huge dam is proposed for the Selangor River. Conservationists fear that the dam may affect water flow, river level and salinity downstream, spelling doom for the fireflies. Opposition to the dam is being loudly voiced, not only by environmentalists but also by those kampung folk whose livelihood depends upon the tourist dollar. Thus, it may be that the fireflies need the tourists in order to survive.
Next Christmas, instead of following the hordes of people to view the annual light-up of Singapore's Orchard Road, why not take a quiet trip up-country to view the unique, world-renowned fireflies of Kuala Selangor. It will be an experience of a lifetime.
The original text for this article first appeared in Volume 8, Number 3 of Nature Watch, published by the Nature Society (Singapore), September 2000


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For sale in Seattle
$275K Gets You a Whidbey Island Log Cabin on Five Acres
Tired of being in the middle of crowded neighborhoods and sirens? Tiny houses on big lots are trending, like this iconic log cabin on Whidbey Island.
By Tom Trimbath@tetrimbath Apr 14, 2016, 8:00am PDT
There's a meme going about that appeals to people who want a quiet and small house on a big lot in the woods, rather than an urban life. Up on Whidbey is a 1 bedroom, 1 bath, 1,139 square foot log cabin on 5.6 acres for $275,000 that might match that dream.
The log cabin was built in 2005, but looks like a classic cabin. Big, rough-hewn logs, stacked into exterior walls, with bigger logs acting as beams and pillars supporting the roof and floors. The floor is stone tile, which makes most sense around the petite wood stove. Upstairs in the loft is the single bedroom. Downstairs is a bathroom with a claw foot tub and similar fixtures that maintain the image of an old-timey cabin.
The land emphasizes the rural character, as it should. With three pastures, and attendant out buildings, you have enough room for horses, livestock, or really big pets. The land is sloped, which helps keep the fields from becoming mud; and tended well with enough trees left to act as natural landscaping.
Tying the house to the land are a covered porch for sitting and enjoying the view, and an uncovered patio that appears to be equipped with an outdoor brick oven for foodies who like baking and roasting without heating up or smoking out the kitchen. If the weather's too cool, but you need some sunshine and warmth, a solarium/greenhouse lets you and your plants soak up photons without getting soaked by the rain.
With that much wood and nature around, you might want to check for termites and other pests. That's something you don't have to worry about if you stick to a new condo in downtown Seattle.
· 4619 Smugglers Cove Rd, Greenbank [Windermere]