lec15_lebon_putz

lec15_lebon_putz - Soluble CPG15 expressed during early...

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Soluble CPG15 expressed during early development rescues cortical progenitors from apoptosis Putz, Harwell, Nedivi, Nature Neuroscience, March 2005 Lauren LeBon April 5, 2005 9.12
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What we knew An activity-dependent protein, cpg15, had been identified in late development to regulate growth of dendrites and axons, and promoting maturation, making it a player in synaptic plasticity and the “critical periods” of development. However, the protein was also known to be present pre-natally, though its role then was unknown. What is the protein doing at this stage? We also knew that apoptosis was occuring early in development, but the survival factors controlling this cell death were unknown. Question : Is cpg15 a survival factor--that is, does it interrupt the apoptotic pathway-- in early development?
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Experimental Strategy Perform in-situ Is cpg15 in the right time at the right place? screens at different stages of development. Could the cpg15 protein reach many different cells? Test cpg15 protein for its ability to diffuse outside the cell of origin. Is cpg15 necessary and sufficient to rescue cells from apoptosis? Harvest soluble cpg15 and place it in a culture of starving neurons, and see if the neurons are rescued from apoptosis. Will cpg15 silencing cause increased progenitor apoptosis in vitro AND in vivo? Knock out cpg15 mRNA with RNAi method in culture and in vivo, assay for progenitor number and apoptosis. Will cpg15 overexpression cause an increase in the progenitor pool? Will it do this by reducing apoptosis? Inject virus containing cpg15 into developing cortex. Assay for progenitor number, cell mitosis, and apoptosis.
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cpg15 : right place, right time At the earliest stages, E14 and E15, cpg15 is present in the cortical plate, the ventricular zone of the dorsal thalamus, and the RGCs, correlating with the proliferation of progenitors and high apoptosis of unnecessary neuroblasts. At E17-E19, cpg15 is expressed in SZV of the telencephelon and the diencephelon and the hippocampal primordia (Fig 1g- l), also correlating with the expansion of the progenitor pool in those regions. Source: Putz, U., C. Harwell, et al. "Soluble CPG15 Expressed During Early Development Rescues Cortical Progenitors From Apoptosis." Nature Neuroscience 8, no. 3 (2005): 322-31. Courtesy of the authors. Used with permission.
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cpg15 : right place, right time At postnatal day 7, cpg15 is expressed in the external granular layers of the cerebellum (Fig. m-n, sagittal sections.). Also note that cpg15 expression returns to the cortex postnatally, this time in the diff. layers. This correlates with the emergence of activity- dependent plasticity in this area.
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