This project will exploit the power of cellular and genetic approaches in zebrafish to illuminate the functions of new genes that are essential for Schwann cell development and myelination in the vertebrate nervous system. In the peripheral nervous system, Schwann cells form the myelin sheath that wraps axons and allows for the rapid transmission of action potentials. Disruption of myelin causes peripheral neuropathies, debilitating diseases that affect 1.5 million people in the United States. Gaps in the understanding of the signals that govern the formation of myelin have hindered the development of therapies for the repair of myelinated axons. In genetic screens for mutations that disrupt myelination, we identified 13 mutations in 10 different genes with specific functions in the development of myelinated axons. Cellular and molecular studies demonstrate that the mutations define genes that function at many steps of the development of myelinated axons, including glial fate specification, Schwann cell migration, organization of the nodes of Ranvier, transport of specific myelin mRNAs within myelinating glia, and commitment of Schwann cells to myelination. In this application, we focus on a group of genes that have key functions in Schwann cell development and myelination. (1) In previous studies, we have identified mutations in erbb2 and erbb3, genes that encode components of a heteromeric receptor for Neuregulin (Nrg) signals, and showed that ErbB signaling is essential for directed migration of Schwann cells. In the present application, we propose to identify the operative signals and determine how they direct Schwann cells as they migrate along axons of growing peripheral nerves. We propose to determine which Nrg isoforms guide Schwann cell migration, to determine where these signals are expressed in developing nerves, and to determine whether Nrg signals are sufficient to guide migrating Schwann cells to ectopic locations. (2) In recent work, we found that two of our mutations disrupt a novel transmembrane protein. We propose to determine whether the protein acts as a signal or a receptor, or both, and to test the hypothesis that the gene acts in neurons to instruct Schwann cells to initiate myelination. In addition, we will test the hypothesis that this new transmembrane protein triggers myelination by activating krox20 in Schwann cells. (3) Our preliminary studies show that the st64 gene, which we are working to identify by positional cloning, is required for Schwann cell myelination. Our characterization suggests that analysis of the st64 mutation will define the function of a novel gene with an essential role in Schwann cell development. We propose to define the cellular and biochemical functions of the st64 gene by phenotypic analysis of the mutants and molecular analysis of the mutated gene. PUBLIC HEALTH RELEVANCE: Disruption of the myelin sheath causes peripheral neuropathies, debilitating diseases that affect 1.5 million people in the United States. The long-term goal of this project is to discover new genes with essential functions in myelin formation using the powerful genetic and cellular approaches available in the zebrafish. The discovery of new genes and the development of new animal models will be an important step toward myelin repair therapies for peripheral neuropathies and other diseases of myelin.