Ca2+-dependent activator protein for secretion 1 is critical for constitutive and regulated exocytosis but not for loading of transmitters into dense core vesicles.
Although CAPS1 was originally identified as a soluble factor that reconstitutes Ca(2+)-dependent secretion from permeabilized neuroendocrine cells, its exact function in intact mammalian cells remains controversial. Here we investigate the role for CAPS1 by generating stable cell lines in which CAPS1 is strongly down-regulated. In these cells, Ca(2+)-dependent secretion was strongly reduced not only of catecholamine but also of a transfected neuropeptide. These secretion defects were rescued by infusion of CAPS1-containing brain cytosol or by transfection-mediated expression of CAPS1. Whole cell patch clamp recording revealed significant reductions in slow burst and sustained release components of exocytosis in the knockdown cells. Unexpectedly, they also accumulated higher amounts of endogenous and exogenous transmitters, which were attributable to reductions in constitutive secretion. Electron microscopy did not reveal abnormalities in the number or docking of dense core vesicles. Our results indicate that CAPS1 plays critical roles not only in Ca(2+)-dependent, regulated exocytosis but also in constitutive exocytosis downstream of vesicle docking. However, they do not support the role for CAPS1 in loading transmitters into dense core vesicles.
Fujita
Yoshihito
Y
Division of Fundamental Neurobiology, Toronto Western Research Institute, University Health Network, Department of Physiology, University of Toronto, Toronto, Ontario M5T 2S8, Canada.
Xu
Ainan
A
Xie
Li
L
Arunachalam
Lakshmanan
L
Chou
Ting-Chieh
TC
Jiang
Tiandan
T
Chiew
Soon-Kwang
SK
Kourtesis
John
J
Wang
Li
L
Gaisano
Herbert Y
HY
Sugita
Shuzo
S
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D019439:Chromaffin Cells / Q000166:cytology; D015536:Down-Regulation; D005089:Exocytosis; D051379:Mice; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D008954:Models, Biological; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009479:Neuropeptides / Q000737:chemistry; D016716:PC12 Cells; D018408:Patch-Clamp Techniques; D051381:Rats; D022142:Secretory Vesicles / Q000378:metabolism*
C510820:Cadps protein, mouse; C485812:Cadps protein, rat; D002135:Calcium-Binding Proteins; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D002118:Calcium