Phospholipase C-eta 2 is required for retinoic acid-stimulated neurite growth

Petra Popovics, Alexander Gray, Mohammed Arastoo, Deana K. Finelli, Audrey J. L. Tan, Alan J. Stewart

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    6 Citations (Scopus)

    Abstract

    Phospholipase C-2 is a recently identified phospholipase C (PLC) implicated in the regulation of neuronal differentiation/maturation. PLC2 activity is triggered by intracellular calcium mobilization and likely serves to amplify Ca2+ signals by stimulating further Ca2+ release from Ins(1,4,5)P3-sensitive stores. The role of PLC2 in neuritogenesis was assessed during retinoic acid (RA)-induced Neuro2A cell differentiation. PLC2 expression increased two-fold during a 4-day differentiation period. Stable expression of PLC2-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 PLC2 knockdown. Using a bacterial two-hybrid screen, we identified LIM domain kinase 1 (LIMK1) as a putative interaction partner of PLC2. Immunostaining of PLC2 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 PLC2 knock-down cells. The phosphoinositide-binding properties of the PLC2 PH domain, assessed using a FRET-based assay, revealed this domain to possess a high affinity toward PtdIns(3,4,5)P3. Immunostaining of PLC2 together with PtdIns(3,4,5)P3 in the Neuro2A cells revealed a high degree of co-localization, indicating that PtdIns(3,4,5)P3 levels in cellular compartments are likely to be important for the spatial control of PLC2 signaling.

    Original languageEnglish
    Pages (from-to)632-644
    Number of pages13
    JournalJournal of Neurochemistry
    Volume124
    Issue number5
    DOIs
    Publication statusPublished - 2013

    Keywords

    • LIM-KINASE
    • BINDING
    • GENE-EXPRESSION
    • INOSITOL 1,4,5-TRISPHOSPHATE
    • MOLECULAR-CLONING
    • OUTGROWTH
    • phospholipase
    • calcium signaling
    • NEUROBLASTOMA-CELLS
    • neurite growth
    • neuroscience
    • nuclear receptors
    • DEPENDENT PROTEIN-KINASE
    • NEURONAL DIFFERENTIATION
    • cell differentiation
    • NUCLEUS

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