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In JoVE (1)
Other Publications (1)
Articles by Nicole Gruenig in JoVE
JoVE Neuroscience
Efficient Gene Delivery into Multiple CNS Territories Using In Utero Electroporation
1Department of Biochemistry and Molecular Biology, Hotchkiss Brain Institute, Alberta Children’s Hospital Research Institute, University of Calgary, 2Department of Medical Genetics, Alberta Children’s Hospital Research Institute, Hotchkiss Brain Institute, University of Calgary
In utero electroporation allows for rapid gene delivery in a spatially- and temporally-controlled manner in the developing central nervous system (CNS). Here we describe a highly adaptable in utero electroporation protocol that can be used to deliver expression constructs into multiple embryonic CNS domains, including the telencephalon, diencephalon and retina.
Other articles by Nicole Gruenig on PubMed
βPix Plays a Dual Role in Cerebral Vascular Stability and Angiogenesis, and Interacts with Integrin α(v)β(8)
The growth of new blood vessels by angiogenesis and their stabilization by the recruitment of perivascular mural cells are thought to be two sequential, yet independent events. Here we identify molecular links between both processes through the βPix and integrin α(v)β(8) proteins. Bubblehead (bbh) mutants with a genetic mutation in βPix show defective vascular stabilization. βPix is a guanine nucleotide exchange factor and scaffold protein that binds many proteins including Git1, which bridges βPix to integrins at focal adhesions. Here we show that the ability of βPix to stabilize vessels requires Git1 binding residues. Knockdown of Git1 leads to a hemorrhage phenotype similar to loss of integrin α(v), integrin β(8) or βPix, suggesting that vascular stabilization through βPix involves interactions with integrins. Furthermore, double loss of function of βPix and integrin α(v) shows enhanced hemorrhage rates. Not only is vascular stability impaired in these embryos, but we also uncover a novel role of both βPix and integrin α(v)β(8) in cerebral angiogenesis. Downregulation of either βPix or integrin α(v)β(8) results in fewer and morphologically abnormal cerebral arteries penetrating the hindbrain. We show that this is coupled with a significant reduction in endothelial cell proliferation in bbh mutants or integrin α(v)β(8) morphants. These data suggest that a complex involving βPix, GIT1 and integrin α(v)β(8) may regulate vascular stability, cerebral angiogenesis and endothelial cell proliferation in the developing embryo.
