PMID- 21187285 OWN - NLM STAT- MEDLINE DCOM- 20110511 LR - 20211020 IS - 1083-351X (Electronic) IS - 0021-9258 (Print) IS - 0021-9258 (Linking) VI - 286 IP - 10 DP - 2011 Mar 11 TI - Phospholipase Cdelta3 regulates RhoA/Rho kinase signaling and neurite outgrowth. PG - 8459-8471 LID - S0021-9258(20)53988-4 [pii] LID - 10.1074/jbc.M110.171223 [doi] AB - Phospholipase Cdelta3 (PLCdelta3) is a key enzyme regulating phosphoinositide metabolism; however, its physiological function remains unknown. Because PLCdelta3 is highly enriched in the cerebellum and cerebral cortex, we examined the role of PLCdelta3 in neuronal migration and outgrowth. PLCdelta3 knockdown (KD) inhibits neurite formation of cerebellar granule cells, and application of PLCdelta3KD using in utero electroporation in the developing brain results in the retardation of the radial migration of neurons in the cerebral cortex. In addition, PLCdelta3KD inhibits axon and dendrite outgrowth in primary cortical neurons. PLCdelta3KD also suppresses neurite formation of Neuro2a neuroblastoma cells induced by serum withdrawal or treatment with retinoic acid. This inhibition is released by the reintroduction of wild-type PLCdelta3. Interestingly, the H393A mutant lacking phosphatidylinositol 4,5-bisphosphate hydrolyzing activity generates supernumerary protrusions, and a constitutively active mutant promotes extensive neurite outgrowth, indicating that PLC activity is important for normal neurite outgrowth. The introduction of dominant negative RhoA (RhoA-DN) or treatment with Y-27632, a Rho kinase-specific inhibitor, rescues the neurite extension in PLCdelta3KD Neuro2a cells. Similar effects were also detected in primary cortical neurons. Furthermore, the RhoA expression level was significantly decreased by serum withdrawal or retinoic acid in control cells, although this decrease was not observed in PLCdelta3KD cells. We also found that exogenous expression of PLCdelta3 down-regulated RhoA protein, and constitutively active PLCdelta3 promotes the RhoA down-regulation more significantly than PLCdelta3 upon differentiation. These results indicate that PLCdelta3 negatively regulates RhoA expression, inhibits RhoA/Rho kinase signaling, and thereby promotes neurite extension. FAU - Kouchi, Zen AU - Kouchi Z AD - From the Laboratory of Genome and Biosignal, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 192-0392 Tokyo. FAU - Igarashi, Takahiro AU - Igarashi T AD - From the Laboratory of Genome and Biosignal, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 192-0392 Tokyo. FAU - Shibayama, Nami AU - Shibayama N AD - From the Laboratory of Genome and Biosignal, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 192-0392 Tokyo. FAU - Inanobe, Shunichi AU - Inanobe S AD - From the Laboratory of Genome and Biosignal, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 192-0392 Tokyo. FAU - Sakurai, Kazuyuki AU - Sakurai K AD - From the Laboratory of Genome and Biosignal, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 192-0392 Tokyo. FAU - Yamaguchi, Hideki AU - Yamaguchi H AD - From the Laboratory of Genome and Biosignal, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 192-0392 Tokyo,; PRESTO, Japan Science and Technology Agency, 4-1-8 Honcho Kawaguchi, Saitama, and. FAU - Fukuda, Toshifumi AU - Fukuda T AD - the Laboratory of Molecular Biochemistry, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 192-0392 Tokyo, Japan. FAU - Yanagi, Shigeru AU - Yanagi S AD - the Laboratory of Molecular Biochemistry, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 192-0392 Tokyo, Japan. FAU - Nakamura, Yoshikazu AU - Nakamura Y AD - From the Laboratory of Genome and Biosignal, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 192-0392 Tokyo. FAU - Fukami, Kiyoko AU - Fukami K AD - From the Laboratory of Genome and Biosignal, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 192-0392 Tokyo,. Electronic address: kfukami@ls.toyaku.ac.jp. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20101227 PL - United States TA - J Biol Chem JT - The Journal of biological chemistry JID - 2985121R RN - 0 (Amides) RN - 0 (Enzyme Inhibitors) RN - 0 (Nerve Tissue Proteins) RN - 0 (Phosphatidylinositol 4,5-Diphosphate) RN - 0 (Pyridines) RN - 138381-45-0 (Y 27632) RN - EC 2.7.11.14 (G-Protein-Coupled Receptor Kinase 1) RN - EC 2.7.11.14 (Grk1 protein, mouse) RN - EC 3.1.4.11 (Phospholipase C delta) RN - EC 3.1.4.11 (Plcd3 protein, mouse) RN - EC 3.6.5.2 (RhoA protein, mouse) RN - EC 3.6.5.2 (rho GTP-Binding Proteins) RN - EC 3.6.5.2 (rhoA GTP-Binding Protein) SB - IM MH - Amides/pharmacology MH - Animals MH - Cell Line, Tumor MH - Cerebellum/enzymology MH - Cerebral Cortex/enzymology MH - Down-Regulation/drug effects/*physiology MH - Enzyme Inhibitors/pharmacology MH - G-Protein-Coupled Receptor Kinase 1/genetics/*metabolism MH - Gene Expression Regulation, Enzymologic/drug effects/*physiology MH - Mice MH - Mice, Inbred ICR MH - Mutation, Missense MH - Nerve Tissue Proteins/genetics/*metabolism MH - Neurites/*enzymology MH - Phosphatidylinositol 4,5-Diphosphate/genetics/metabolism MH - Phospholipase C delta/genetics/*metabolism MH - Pyridines/pharmacology MH - Signal Transduction/drug effects/*physiology MH - rho GTP-Binding Proteins/antagonists & inhibitors/*biosynthesis/genetics MH - rhoA GTP-Binding Protein PMC - PMC3048730 EDAT- 2010/12/29 06:00 MHDA- 2011/05/12 06:00 PMCR- 2012/03/11 CRDT- 2010/12/29 06:00 PHST- 2010/12/29 06:00 [entrez] PHST- 2010/12/29 06:00 [pubmed] PHST- 2011/05/12 06:00 [medline] PHST- 2012/03/11 00:00 [pmc-release] AID - S0021-9258(20)53988-4 [pii] AID - M110.171223 [pii] AID - 10.1074/jbc.M110.171223 [doi] PST - ppublish SO - J Biol Chem. 2011 Mar 11;286(10):8459-8471. doi: 10.1074/jbc.M110.171223. Epub 2010 Dec 27.