Synaptotagmin-7 enhances calcium-sensing of chromaffin cell granules and slows discharge of granule cargos.
Acetylcholine
/ pharmacology
Animals
Calcium Signaling
/ genetics
Cell Movement
/ genetics
Chromaffin Granules
/ physiology
Electrophysiological Phenomena
Exocytosis
Female
Kinetics
Male
Membrane Fusion
Mice
Mice, Inbred C57BL
Mice, Knockout
PC12 Cells
Rats
Receptors, Calcium-Sensing
/ physiology
SNARE Proteins
/ metabolism
Subcellular Fractions
/ metabolism
Synaptotagmin I
/ physiology
Synaptotagmins
/ genetics
chromaffin cell
exocytosis
fusion
secretion
synaptotagmin
Journal
Journal of neurochemistry
ISSN: 1471-4159
Titre abrégé: J Neurochem
Pays: England
ID NLM: 2985190R
Informations de publication
Date de publication:
09 2020
09 2020
Historique:
received:
16
07
2019
revised:
04
02
2020
accepted:
07
02
2020
pubmed:
15
2
2020
medline:
10
2
2021
entrez:
15
2
2020
Statut:
ppublish
Résumé
Synaptotagmin-7 (Syt-7) is one of two major calcium sensors for exocytosis in adrenal chromaffin cells, the other being synaptotagmin-1 (Syt-1). Despite a broad appreciation for the importance of Syt-7, questions remain as to its localization, function in mediating discharge of dense core granule cargos, and role in triggering release in response to physiological stimulation. These questions were addressed using two distinct experimental preparations-mouse chromaffin cells lacking endogenous Syt-7 (KO cells) and a reconstituted system employing cell-derived granules expressing either Syt-7 or Syt-1. First, using immunofluorescence imaging and subcellular fractionation, it is shown that Syt-7 is widely distributed in organelles, including dense core granules. Total internal reflection fluorescence (TIRF) imaging demonstrates that the kinetics and probability of granule fusion in Syt-7 KO cells stimulated by a native secretagogue, acetylcholine, are markedly lower than in WT cells. When fusion is observed, fluorescent cargo proteins are discharged more rapidly when only Syt-1 is available to facilitate release. To determine the extent to which the aforementioned results are attributable purely to Syt-7, granules expressing only Syt-7 or Syt-1 were triggered to fuse on planar supported bilayers bearing plasma membrane SNARE proteins. Here, as in cells, Syt-7 confers substantially greater calcium sensitivity to granule fusion than Syt-1 and slows the rate at which cargos are released. Overall, this study demonstrates that by virtue of its high affinity for calcium and effects on fusion pore expansion, Syt-7 plays a central role in regulating secretory output from adrenal chromaffin cells.
Identifiants
pubmed: 32058590
doi: 10.1111/jnc.14986
pmc: PMC7426247
mid: NIHMS1571416
doi:
Substances chimiques
Receptors, Calcium-Sensing
0
SNARE Proteins
0
Synaptotagmin I
0
Syt7 protein, rat
0
Synaptotagmins
134193-27-4
Acetylcholine
N9YNS0M02X
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
598-617Subventions
Organisme : NIGMS NIH HHS
ID : P01 GM072694
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM007767
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32GM007767
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM111997
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01GM111997
Pays : United States
Organisme : NIGMS NIH HHS
ID : P01GM072694
Pays : United States
Organisme : NIGMS NIH HHS
ID : P01GM072694
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01GM111997
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32GM007767
Pays : United States
Informations de copyright
© 2020 International Society for Neurochemistry.
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