Mechanisms controlling neurite outgrowth in a pheochromocytoma cell line

The role of TRPC channels

Sanjay Kumar, Saikat Chakraborty, Cindy Barbosa, Tatiana Brustovetsky, Nikolai Broustovetski, Alexander Obukhov

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Transient Receptor Potential Canonical (TRPC) channels are implicated in modulating neurite outgrowth. The expression pattern of TRPCs changes significantly during brain development, suggesting that fine-tuning TRPC expression may be important for orchestrating neuritogenesis. To study how alterations in the TRPC expression pattern affect neurite outgrowth, we used nerve growth factor (NGF)-differentiated rat pheochromocytoma 12 (PC12) cells, a model system for neuritogenesis. In PC12 cells, NGF markedly up-regulated TRPC1 and TRPC6 expression, but down-regulated TRPC5 expression while promoting neurite outgrowth. Overexpression of TRPC1 augmented, whereas TRPC5 overexpression decelerated NGF-induced neurite outgrowth. Conversely, shRNA-mediated knockdown of TRPC1 decreased, whereas shRNA-mediated knockdown of TRPC5 increased NGF-induced neurite extension. Endogenous TRPC1 attenuated the anti-neuritogenic effect of overexpressed TRPC5 in part by forming the heteromeric TRPC1-TRPC5 channels. Previous reports suggested that TRPC6 may facilitate neurite outgrowth. However, we found that TRPC6 overexpression slowed down neuritogenesis, whereas dominant negative TRPC6 (DN-TRPC6) facilitated neurite outgrowth in NGF-differentiated PC12 cells. Consistent with these findings, hyperforin, a neurite outgrowth promoting factor, decreased TRPC6 expression in NGF-differentiated PC12 cells. Using pharmacological and molecular biological approaches, we determined that NGF up-regulated TRPC1 and TRPC6 expression via a p75NTR-IKK2-dependent pathway that did not involve TrkA receptor signaling in PC12 cells. Similarly, NGF up-regulated TRPC1 and TRPC6 via an IKK2 dependent pathway in primary cultured hippocampal neurons. Thus, our data suggest that a balance of TRPC1, TRPC5, and TRPC6 expression determines neurite extension rate in neural cells, with TRPC6 emerging as an NGF-dependent "molecular damper" maintaining a submaximal velocity of neurite extension.

Original languageEnglish
Pages (from-to)1408-1419
Number of pages12
JournalJournal of Cellular Physiology
Volume227
Issue number4
DOIs
StatePublished - Apr 2012

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Transient Receptor Potential Channels
PC12 Cells
Nerve Growth Factor
Cells
Pheochromocytoma
Neurites
Small Interfering RNA
trkA Receptor
Neuronal Outgrowth
Nerve Growth Factors
Neurons
Rats
Brain
Tuning
Pharmacology

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Mechanisms controlling neurite outgrowth in a pheochromocytoma cell line : The role of TRPC channels. / Kumar, Sanjay; Chakraborty, Saikat; Barbosa, Cindy; Brustovetsky, Tatiana; Broustovetski, Nikolai; Obukhov, Alexander.

In: Journal of Cellular Physiology, Vol. 227, No. 4, 04.2012, p. 1408-1419.

Research output: Contribution to journalArticle

Kumar, Sanjay ; Chakraborty, Saikat ; Barbosa, Cindy ; Brustovetsky, Tatiana ; Broustovetski, Nikolai ; Obukhov, Alexander. / Mechanisms controlling neurite outgrowth in a pheochromocytoma cell line : The role of TRPC channels. In: Journal of Cellular Physiology. 2012 ; Vol. 227, No. 4. pp. 1408-1419.
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