TC10β/CDC42 GTPase activating protein is required for the growth of cortical neuron dendrites

• Published in: Neuroscience Vol. 199; pp. 589 - 597
• Main Authors: Shen, P.-C, Xu, D.-F, Liu, J.-W, Li, K, Lin, M, Wang, H.-T, Wang, R, Zheng, J
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Ltd 29.12.2011; Elsevier
• Subjects: Animals; axonal morphogenesis; Biological and medical sciences; Blotting, Western; Cerebral Cortex - growth & development; Cerebral Cortex - metabolism; Cerebral Cortex - ultrastructure; cortical neurons; dendrite elaboration; Dendrites - metabolism; Dendrites - ultrastructure; dendritic growth; Electroporation; Fundamental and applied biological sciences. Psychology; GTPase-Activating Proteins - genetics; GTPase-Activating Proteins - metabolism; HEK293 Cells; Humans; Immunohistochemistry; Neurogenesis - physiology; Neurology; Neurons - metabolism; Neurons - ultrastructure; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; spine formation; TCGAP; Transfection; Vertebrates: nervous system and sense organs; TCGAP-WT; HRP; PFA; human cytomegalovirus; BSA; shRNA; FBS; RT-PCR; TC10β/CDC42 GTPase activating protein; paraformaldehyde; pregnant embryonic day; PBS; horseradish-peroxidase; postnatal day 7; reverse transcription PCR; NGF; CMV; short-hairpin RNA; fetal bovine serum; Sprague–Dawley; spine formation; days in vitro; dendrite elaboration; GAPDH; bovine serum albumin; PCR; TCGAP-Res; wild-type TCGAP; E; Institute of Cancer Research; DMEM; SD; human embryonic kidney; nerve growth factor; TCGAP; PDL; polymerase chain reaction; shRNA-scr; scrambled oligonucleotide sequence; dendritic growth; glyceraldehydes-3-phosphate dehydrogenase; phosphate buffered-saline; P7; axonal morphogenesis; HEK; DIV; cortical neurons; poly- d-lysine; resistant form of TCGAP; ICR; Dulbecco modified Eagle's medium; Enzyme; Triphosphoric monoester hydrolases; Growth; dGTPase; Esterases; Protein; Dendrite; Morphogenesis; Neuron; Hydrolases
• ISSN: 0306-4522; 1873-7544
• DOI: 10.1016/j.neuroscience.2011.08.053

Abstract Neuronal morphogenesis plays an important role in neuronal development. TC10β/CDC42 GTPase-activating protein (TCGAP) is known to be a brain-enriched multiple domain protein, but its role in neuronal development process remains poorly understood. In the present study, we showed that TCGAP positively regulated dendritic outgrowth and spine formation in developing cortical neurons. Knocking down TCGAP by RNA interference led to a decrease in the overall length of dendrite arbors and the number of dendrite branches both in vitro and in vivo . Overexpressing TCGAP in cultured cortical neurons increased dendritic outgrowth and branching. Moreover, overexpressing TCGAP lead to an increase of spine density while knocking-down TCGAP decreased spine density in vivo . The defect by downregulating TCGAP could be rescued by expressing a knock-down resistant form of TCGAP both in vivo and in vitro . In contrast, neither downregulating nor overexpressing TCGAP had any effect on axonal morphogenesis in primary cortical neuron cultures. Together, our findings suggest that TCGAP regulates neuronal morphogenesis in developing cortical neurons at both early and late stage.