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 language | English |
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Pages (from-to) | 632-644 |
Number of pages | 13 |
Journal | Journal of Neurochemistry |
Volume | 124 |
Issue number | 5 |
DOIs | |
Publication status | Published - 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