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The Journal of Neuroscience, April 1, 2002, 22(7):2427-2433

Alterations in Exocytosis Induced by Neuronal Ca2+ Sensor-1 in Bovine Chromaffin Cells

Chien-Yuan Pan1, Andreas Jeromin2, Kenneth Lundstrom3, Seung Hyun Yoo4, John Roder2, and Aaron P. Fox1

1 Department of Neurobiology, Pharmacology, and Physiology, The University of Chicago, Chicago, Illinois 60637, 2 Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, M5G 1X5 Canada, 3 F. Hoffmann-La Roche, CNS Department, CH-4070 Basel, Switzerland, and 4 National Creative Research Initiative Center for Secretory Granule Research, Korean Advanced Institute of Science and Technology, Dae Jeon 305-701, Korea

A variety of Ca2+ binding proteins are known to play an integral role in catecholamine release from synapses as well as secretory cells, such as chromaffin cells. The Drosophila protein frequenin and its mammalian homolog neuronal Ca2+ sensor-1 (NCS-1) belong to a family of Ca2+ sensors with EF hands that bind Ca2+ and then interact with other proteins. Frequenin/NCS-1 has been shown to enhance exocytotic activity in addition to altering Ca2+ channel regulation. To better understand how NCS-1 regulates stimulus-secretion coupling, bovine chromaffin cells were infected with Semliki Forest virus (SFV) vectors containing the rat NCS-1 gene. Cells were studied in the perforated whole-cell patch-clamp configuration. Membrane capacitance was monitored as an indicator of exocytosis-endocytosis. Exocytosis elicited by membrane depolarization was not significantly different between cells infected with SFV expressing green fluorescent protein (GFP) or GFP plus NCS-1, except that the overexpression of NCS-1 resulted in a faster rundown in exocytosis. When cells were stimulated with histamine, NCS-1 overexpression led to higher exocytosis, as well as [Ca2+]i elevation. Immunocytochemistry showed a similar distribution of NCS-1 and phosphatidylinositol 4-kinase beta  (PI4Kbeta ). NCS-1 and PI4Kbeta coimmunoprecipitate, opening up the possibility that the two proteins directly interact. These results suggest that NCS-1 may regulate cellular activity through the modulation of the phosphatidylinositol signaling pathway.

Key words: neuronal calcium sensor; NCS-1; chromaffin; exocytosis; calcium; histamine; phosphatidylinositol; IP3; Semliki Forest virus vector


Copyright © 2002 Society for Neuroscience  0270-6474/02/2272427-07$05.00/0


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