Modulation of muscarinic signaling in PC12 cells overexpressing neuronal Ca2+ sensor-1 protein
Corresponding Author(s) : M M Guimaràes
Cellular and Molecular Biology,
Vol. 55 No. 4: General Papers
It has been suggested that overexpression of neuronal Ca2+ sensor-1 (NCS-1) protein is implicated in the pathophysiology of neurodisorders such as schizophrenia, bipolar disturbance and X-linked mental retardation. The mechanism by which NCS-1 would be involved in the causes and/or consequences of these neurodisorders is still far from elucidation. Independent evidence has pointed NCS-1 as a key regulator of synaptic efficacy by altering the expression and activity of voltage-gated channels, inhibiting internalization of dopaminergic receptors, and altering phosphoinositide metabolism. In this study, we examined the possible participation of NCS-1 protein in signal transmission dependent on muscarinic receptor activation, using PC12 cells stably expressing NCS-1 (PC12-NCS-1). Carbachol (CCH; 300µM) was able to evoke glutamate release more efficiently from PC12-NCS-1 (15.3±1.0nmol/mg of protein) than wild type cells (PC12-wt; 8.3±0.9nmol/mg of protein). This increase of glutamate release induced by CCH was independent on extracellular Ca2+ influx. Additionally, a larger increase of cytoplasmic levels of InsP3 (663.0±63.0 and 310.0±39.0% of fluorescence in A.U.) and [Ca2+]i (766.4±40.0 and 687.8±37.1nmol/L) was observed after CCH stimulus of PC12-NCS-1 compared with PC12-wt. Clearly distinction between intracellular Ca2+ dynamics was also observed in PC12-NCS-1 and PC12-wt. A larger increase followed by fast decay of [Ca2+]i was observed in PC12-NCS-1. A plateau with a delayed decay of [Ca2+]i was characteristic of PC12-wt [Ca2+]i response. Both enhancement of InsP3 production and glutamate release observed in PC12-NCS-1 were blocked by atropine (10µM). Together, our data show that overexpression of NCS-1 in PC12 cells induces an enhancement of intracellular second messenger and transmitter release dependent on CCH response, suggesting that muscarinic signaling is "up-regulated” in this cell model.
PC12 cells NCS-1 Calcium signaling glutamate release.
Guimaràes, M. M., Reis, H. J., Guimaràes, L. P., Carneiro, D. S., Ribeiro, F. M., Gomez, M. V., Jeromin, A., & Romano-Silva, M. A. (2009). Modulation of muscarinic signaling in PC12 cells overexpressing neuronal Ca2+ sensor-1 protein. Cellular and Molecular Biology, 55(4), 1138–50. Retrieved from https://cellmolbiol.org/index.php/CMB/article/view/1056
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