to determine the distribution and level of raav9 mediated transgene expression naglu specific if was detected throughout the brains of treated mice in neurons glia and abundant endothelial cells in capillaries and larger blood vessels in an apparently dose u2014 dependent fashion no significant differences were observed in the distribution or levels of rnaglu signal between 6 months and 9 months pi naglu positive glial cells were not costained with anti glial fibrillary acidic protein gfap ab and were likely to be oligodendrocytes based on their morphology importantly while rnaglu if was observed in the brains of all raav9 treated mice mannitol pretreatment did appear to increase the number of transduced cells in the cns differential transduction levels were observed in peripheral organs the rnaglu protein was detected in 20 40 of hepatocytes gt 95 of cardiomyocytes and 10 30 of skeletal myocytes the distribution of raav9 transduced hepatocytes was uniform throughout the liver transduction in abundant neurons in myenteric plexus and submucosal plexus of the intestine was observed suggesting efficient targeting of the peripheral nervous system pns the rnaglu signals were mostly present in granules whereas scaav9 mediated gfp signals were uniform in the cytoplasm of transduced cells suggesting correct lysosomal trafficking of rnaglu transduction of endothelial cells was also observed in peripheral tissues of raav9 gfp vector treated mice function of the recombinant naglu and resulting effects in animals were also analyzed in the therapeutic studies above transgene enzymatic activity was assayed to quantify the expression and the functionality of raav9 mediated rnaglu there were no significant differences in tissue naglu activity.