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Phospholipase C‐η2 is required for retinoic acid‐stimulated neurite growth
Author(s) -
Popovics Petra,
Gray Alexander,
Arastoo Mohammed,
Finelli Deana K.,
Tan Audrey J. L.,
Stewart Alan J.
Publication year - 2013
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/jnc.12122
Subject(s) - retinoic acid , neurite , phospholipase c , microbiology and biotechnology , gene knockdown , immunostaining , signal transduction , chemistry , biology , biochemistry , immunology , in vitro , apoptosis , immunohistochemistry , gene
Phospholipase C‐η2 is a recently identified phospholipase C (PLC) implicated in the regulation of neuronal differentiation/maturation. PLCη2 activity is triggered by intracellular calcium mobilization and likely serves to amplify Ca 2+ signals by stimulating further Ca 2+ release from Ins(1,4,5)P 3 ‐sensitive stores. The role of PLCη2 in neuritogenesis was assessed during retinoic acid (RA)‐induced Neuro2A cell differentiation. PLCη2 expression increased two‐fold during a 4‐day differentiation period. Stable expression of PLCη2‐targetted shRNA led to a decrease in the number of differentiated cells and total length of neurites following RA‐treatment. Furthermore, RA response element activation was perturbed by PLCη2 knockdown. Using a bacterial two‐hybrid screen, we identified LIM domain kinase 1 (LIMK1) as a putative interaction partner of PLCη2. Immunostaining of PLCη2 revealed significant co‐localization with LIMK1 in the nucleus and growing neurites in Neuro2A cells. RA‐induced phosphorylation of LIMK1 and cAMP ‐responsive element‐binding protein was reduced in PLCη2 knock‐down cells. The phosphoinositide‐binding properties of the PLCη2 PH domain, assessed using a FRET‐based assay, revealed this domain to possess a high affinity toward PtdIns(3,4,5)P 3 . Immunostaining of PLCη2 together with PtdIns(3,4,5)P 3 in the Neuro2A cells revealed a high degree of co‐localization, indicating that PtdIns(3,4,5)P 3 levels in cellular compartments are likely to be important for the spatial control of PLCη2 signaling.