Expression of PKC Iota Affects Neuronal Differentiation of PC12 Cells at Least Partly Independent of Kinase Function


Atypical PKC (aPKC) plays a role in establishing cell polarity and has been indicated in neuronal differentiation and polarization, including neurite formation in rat pheochromocytoma PC12 cells, albeit by unclear mechanisms. Here, the role of the aPKC isoform, PKC iota (PKCι), in the early neuronal differentiation of PC12 cells, was investigated. NGF-treated PC12 cells with stably expressed exogenous wild-type PKCι showed decreased expression of a neuroendocrine marker, increased expression of a neuronal marker and increased neurite formation. Stable expression of a kinase-inactive PKCι, but not constitutively active PKCι lacking a regulatory domain, had similar though less potent effects. Pharmacological inhibition of endogenous aPKC kinase activity in parental PC12 cells did not inhibit neurite formation, suggesting that some of the observed effects of PKCι expression on neuronal differentiation are kinase-independent. Interestingly, exogenous expression of wild-type and kinase-inactive PKCι had little effect on overall PKCι activity, but caused a decrease in PKC zeta (PKCζ) kinase activity, suggesting an interplay between the two isoforms that may underlie the observed results. Overall, these findings suggest that in PC12 and perhaps other neuroendocrine precursor cells, PKCι influences an early differentiation decision between the neuroendocrine (chromaffin) and sympathetic neuron cell lineages, potentially by affecting PKCζ function.

Share and Cite:

Doonachar, A. and R. Schoenfeld, A. (2014) Expression of PKC Iota Affects Neuronal Differentiation of PC12 Cells at Least Partly Independent of Kinase Function. CellBio, 3, 1-13. doi: 10.4236/cellbio.2014.31001.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Nishizuka, Y. (1995) Protein Kinase C and Lipid Signaling for Sustained Cellular Responses. The FASEB Journal, 9, 484-496.
[2] Newton, A.C. (2010) Protein Kinase C: Poised to Signal. American Journal of Physiology Endocrinology and Metabolism, 298, E395-E402.
[3] Rosse, C., Linch, M., Kermorgant, S., Cameron, A.J., Boeckeler, K. and Parker, P.J. (2010) PKC and the Control of Localized Signal Dynamics. Nature Reviews Molecular Cell Biology, 11, 103-112.
[4] Izumi, Y., Hirose, T., Tamai, Y., Hirai, S., Nagashima, Y., Fujimoto, T., et al. (1998) An Atypical PKC Directly Associates and Colocalizes at the Epithelial Tight Junction with ASIP, a Mammalian Homologue of Caenorhabditis Elegans Polarity Protein PAR-3. Journal of Cell Biology, 143, 95-106.
[5] Shi, S.H., Jan, L.Y. and Jan, Y.N. (2003) Hippocampal Neuronal Polarity Specified by Spatially Localized mPar3/ mPar6 and PI 3-Kinase Activity. Cell, 112, 63-75.
[6] Nishimura, T., Yamaguchi, T., Kato, K., Yoshizawa, M., Nabeshima, Y., Ohno, S., et al. (2005) PAR-6-PAR-3 Mediates Cdc42-Induced Rac Activation through the Rac GEFs STEF/Tiam1. Nature Cell Biology, 7, 270-277.
[7] Shi, S.H., Cheng, T., Jan, L.Y. and Jan, Y.N. (2004) APC and GSK-3beta Are Involved in mPar3 Targeting to the Nascent Axon and Establishment of Neuronal Polarity. Current Biology, 14, 2025-2032.
[8] Greene, L.A.and Tischler, A.S. (1976) Establishment of a Noradrenergic Clonal Line of Rat Adrenal Pheochromocytoma Cells Which Respond to Nerve Growth Factor. Proceedings of the National Academy of Sciences USA, 73, 2424-2428.
[9] Soh, J.W., Mao, Y., Liu, L., Thompson, W.J., Pamukcu, R. and Weinstein, I.B. (2001) Protein Kinase G Activates the JNK1 Pathway via Phosphorylation of MEKK1. Journal of Biological Chemistry, 276, 16406-16410.
[10] Soh, J.W. and Weinstein, I.B. (2003) Roles of Specific Isoforms of Protein Kinase c in the Transcriptional Control of Cyclin D1 and Related Genes. Journal of Biological Chemistry, 278, 34709-34716.
[11] Naviaux, R.K., Costanzi, E., Haas, M. and Verma, I.M. (1996) The pCL Vector System: Rapid Production of Helper-Free, High-Titer, Recombinant Retroviruses. Journal of Virology, 70, 5701-5705.
[12] Bangiyeva, V., Rosenbloom, A., Alexander, A., Isanova, B., Popko, T. and Schoenfeld, A. (2009) Differences in Regulation of Tight Junctions and Cell Morphology between VHL Mutations from Disease Subtypes. BMC Cancer, 9, 229.
[13] Reissmann, E., Ernsberger, U., Francis-West, P.H., Rueger, D., Brickell, P.M. and Rohrer, H. (1996) Involvement of Bone Morphogenetic Protein-4 and Bone Morphogenetic Protein-7 in the Differentiation of the Adrenergic Phenotype in Developing Sympathetic Neurons. Development, 122, 2079-2088.
[14] Peng, I., Binder, L.I. and Black, M.M. (1985) Cultured Neurons Contain a Variety of Microtubule-Associated Proteins. Brain Research, 361, 200-211.
[15] Rossino, P., Gavazzi, I., Timpl, R., Aumailley, M., Abbadini, M., Giancotti, F., et al. (1990) Nerve Growth Factor Induces Increased Expression of a Laminin-Binding Integrin in Rat Pheochromocytoma PC12 Cells. Experimental Cell Research, 189, 100-108.
[16] Dotti, C.G., Sullivan, C.A. and Banker, G.A. (1988) The Establishment of Polarity by Hippocampal Neurons in Culture. Journal of Neuroscience, 8, 1454-1468.
[17] Ludin, B. and Matus, A. (1993) The Neuronal Cytoskeleton and Its Role in Axonal and Dendritic Plasticity. Hippocampus, 3, 61-71.
[18] Caceres, A., Mautino, J. and Kosik, K.S. (1992) Suppression of MAP2 in Cultured Cerebellar Macroneurons Inhibits Minor Neurite Formation. Neuron, 9, 607-618.
[19] Sharma, N., Kress, Y. and Shafit-Zagardo, B. (1994) Antisense MAP-2 Oligonucleotides Induce Changes in Microtubule Assembly and Neuritic Elongation in Pre-Existing Neurites of Rat Cortical Neurons. Cell Motility and the Cytoskeleton, 27, 234-247.
[20] Coleman, E.S. and Wooten, M.W. (1994) Nerve Growth Factor-Induced Differentiation of PC12 Cells Employs the Pma-Insensitive Protein Kinase C-Zeta Isoform. Journal of Molecular Neuroscience, 5, 39-57.
[21] Wooten, M.W., Seibenhener, M.L., Zhou, G., Vandenplas, M.L. and Tan, T.H. (1999) Overexpression of Atypical PKC in PC12 Cells Enhances NGF-Responsiveness and Survival through an NF-KappaB Dependent Pathway. Cell Death and Differentiation, 6, 753-764.
[22] Gschwendt, M., Dieterich, S., Rennecke, J., Kittstein, W., Mueller, H.J. and Johannes, F.J. (1996) Inhibition of Protein Kinase C Mu by Various Inhibitors. Differentiation from Protein Kinase C Isoenzymes. FEBS Letters, 392, 77-80.
[23] Nagai-Tamai, Y., Mizuno, K., Hirose, T., Suzuki, A. and Ohno, S. (2002) Regulated Protein-Protein Interaction between aPKC and PAR-3 Plays an Essential Role in the Polarization of Epithelial Cells. Genes to Cells, 7, 1161-1171.
[24] Suzuki, A., Yamanaka, T., Hirose, T., Manabe, N., Mizuno, K., Shimizu, M., et al. (2001) Atypical Protein Kinase C Is Involved in the Evolutionarily Conserved Par Protein Complex and Plays a Critical Role in Establishing Epithelia-Specific Junctional Structures. Journal of Cell Biology, 152, 1183-1196.
[25] Joberty, G., Petersen, C., Gao, L. andMacara, I.G. (2000) The Cell-Polarity Protein Par6 Links Par3 and Atypical Protein Kinase C to Cdc42. Nature Cell Biology, 2, 531-539.
[26] Lin, D., Edwards, A.S., Fawcett, J.P., Mbamalu, G., Scott, J.D. and Pawson, T. (2000) A Mammalian PAR-3-PAR-6 Complex Implicated in Cdc42/Rac1 and aPKC Signalling and Cell Polarity. Nature Cell Biology, 2, 540-547.
[27] Zhou, J., Fariss, R.N. and Zelenka, P.S. (2003) Synergy of Epidermal Growth Factor and 12(S)-Hydroxyeicosatetraenoate on Protein Kinase C Activation in Lens Epithelial Cells. Journal of Biological Chemistry, 278, 5388-5398.
[28] Daniels, R.H., Hall, P.S. and Bokoch, G.M. (1998) Membrane Targeting of p21-Activated Kinase 1 (PAK1) Induces Neurite Outgrowth from PC12 Cells. EMBO Journal, 17, 754-764.
[29] Weston, C.A., Anova, L., Rialas, C., Prives, J.M. and Weeks, B.S. (2000) Laminin-1 Activates Cdc42 in the Mechanism of Laminin-1-Mediated Neurite Outgrowth. Experimental Cell Research, 260, 374-378.
[30] Goldstein, B. andMacara, I.G. (2007) The PAR Proteins: Fundamental Players in Animal Cell Polarization. Developmental Cell, 13, 609-622.
[31] Balendran, A., Biondi, R.M., Cheung, P.C., Casamayor, A., Deak, M. and Alessi, D.R. (2000) A 3-Phosphoinositide-Dependent Protein Kinase-1 (PDK1) Docking Site Is Required for the Phosphorylation of Protein Kinase Czeta (PKCzeta) and PKC-Related Kinase 2 by PDK1. Journal of Biological Chemistry, 275, 20806-20813.
[32] Wang, G., Krishnamurthy, K., Umapathy, N.S., Verin, A.D. and Bieberich, E. (2009) The Carboxyl-Terminal Domain of Atypical Protein Kinase C Zeta Binds to Ceramide and Regulates Junction Formation in Epithelial Cells. Journal of Biological Chemistry, 284, 14469-14475.
[33] Pillai, P., Desai, S., Patel, R., Sajan, M., Farese, R., Ostrov, D., et al. (2011) A Novel PKC-Iota Inhibitor Abrogates Cell Proliferation and Induces Apoptosis in Neuroblastoma. International Journal of Biochemistry and Cell Biology, 43, 784-794.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.