CELL BIOLOGY OF THE NEURON Polarity ... - Tavernarakis Lab

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Cell Biology of the Neuron: <strong>Polarity</strong>, Plasticity and Regeneration, Crete 2011 The Oriented Emergence of Axons from Retinal Ganglion Cells is Directed by Laminin Contact in vivo Owen Randlett 1 , Lucia Poggi 2 , Flavio R Zolessi 3 , William A Harris 1 1 Department of Physiology, Development and Neuroscience, University of Cambridge, United Kingdom 2 Institute of Zoology, Heidelberg University, Germany 3 Seccion Biologia Celular, Departamento de Biologia Celular y Molecular, Facultad de Ciencias, Universidad de la Republica, Montevidee, Uruguay The site of axon emergence from the cell body of most differentiating neurons is precisely specified during development. For example, cortical pyramidal neurons send out axons apically. How this is accomplished, and the relative importance of intrinsic and extrinsic mechanisms, is not understood. The axons of retinal ganglion cells (RGCs) emerge basally in vivo, yet because RGCs develop from polarized neuroepithelial cells within a polarized environment, disentangling intrinsic and extrinsic influences is a challenge. We use a combination of in vitro and in vivo time-lapse imaging in zebrafish embryos to demonstrate that Laminin acting directly on RGCs is necessary and sufficient to orient axon emergence in vivo. Laminin contact with the basal processes of newborn RGCs prevents the cells from entering a stochastic Stage 2 phase, directs the rapid accumulation of the early axonal marker Kif5c560-YFP, and leads to the formation of axonal growth cones. These results demonstrate that contact mediated extrinsic cues may be critical for the site of axon emergence, and account for the differences in cellular behavior observed in vitro and in vivo. Presented by: Randlett, Owen 62
Cell Biology of the Neuron: <strong>Polarity</strong>, Plasticity and Regeneration, Crete 2011 Dendrite Morphogenesis and Functional Implications Yuh-Nung Jan , HHMI, UCSF For the past ten years, we have been trying to uncover the rules and the mechanisms that control dendrite morphogenesis by using a group of fly larval sensory neurons known as dendritic arborization (or da) neurons. We have gained some insights about dendrite development including how axons are dendrites are made differently, how a neuron acquires its neuronal type specific morphology, how the dendrites of different neurons are organized, how the size of a dendritic arbor is controlled, and how the pruning and remodeling of dendrites are regulated during development. Of the four different classes of da neurons, the dendrites of class IV da neurons form a regular array that tiles the larval body. Recently we found those neurons actually constitute a novel photoreceptor system using a photo-transduction pathway that is distinct from all the previously known ones. This regular array of photo-sensors enable the larvae to sense light exposure over its entire body to move out of danger. Previously, we found that the Hippo pathway plays important roles in controlling the dendritic arbor size and tiling of class IV da neurons. In Drosophila, Hippo Kinase regulate Tricorner (Trc) and Warts (Wts), the the only two members of Ser/Thr kinases of the NDR family in fly. Each Kinase then regulates complementary aspects of dendritic arbors: Wts controls maintenance where as Trc controls branching and tiling. To see whether our findings could be extended to mice, we study the two Trc homologues, NDR1 and NDR2, in mice. They share high sequence homology and both are ubiquitously expressed in the mouse brain throughout development. We find that NDR1/2 function to limit the complexity of the dendritic arbor of mouse hippocampal and cortical neurons, a role analogous to that of their fly and worm homologues. Additionally, we find a new function for NDR1/2: they are required for the proper development of dendritic spines. In order to gain insights concerning how NDR1/2 exert their functions, we set out to identify the substrates of their kinase activity. In collaboration with our colleague Kevan Shokat’s lab, we were able to apply their ingenious “chemical genetics and covalent capture method” to identify five NDR1/2 substrates and their phosphorylation sites. Strikingly, four have been implicated in vesicle trafficking. We chose two of them, AAK1 and Rabin8, for further studies. We were able not only to validate both as bona fide NDR1/2 kinase substrates but also found that AAK1 is preferentially required for controlling dentritic arbor complexity whereas Rabin8 is preferentially required for dendritic spine maturation – a finding that paves the way for dissecting the NDR1/2 pathway. Presented by: Jan, Yuh-Nung 63
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