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PMID: 8782102 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Hypotonic stimulation induced Ca2+ release from IP3-sensitive internal stores in a green monkey kidney cell line.

The Journal of physiology ·Vol. 493 ( Pt 2) ·1996-06-01 ·Pages 371-84

Ishii T, Hashimoto T, Ohmori H

Abstract

1. Hypotonic stimulation (180 +/- 5 mosmol l-1) increased [Ca2+]i in fura-2-loaded Green monkey kidney cells (COS-7 cells) and depolarized the membrane. 2. COS-7 cells were depolarized up to -3.5 +/- 4.4 mV from a resting membrane potential of -35.2 +/- 2.3 mV in response to hypotonic stimulation, when the patch electrode was filled with a 160 mM KCl-0.5 mM EGTA-based intracellular medium. 3. The increase in [Ca2+]i induced by hypotonic stimulation was divided into two phases. One was transient and oscillatory, and observed in Ca(2+)-free medium; the other was persistent, blocked by 100 microM La3+, and observed only in Ca(2+)-containing medium. 4. The increase in [Ca2+]i in Ca(2+)-free medium was blocked by pretreatment with 10 microM thapsigargin. The increase in [Ca2+]i induced by 10 microM thapsigargin was reduced after hypotonic stimulation which induced an increase in [Ca2+]i in Ca(2+)-free medium. 5. The increase in [Ca2+]i in Ca(2+)-free medium was not affected by treatment with 5 mM caffeine or 1-10 microM ryanodine. Neither caffeine nor ryanodine induced an increase in [Ca2+]i. 6. Adenosine 5'-O-2-thiodiphosphate (ADP-beta-S; a P2Y receptor agonist) induced an increase in [Ca2+]i in Ca(2+)-free medium and caused phosphoinositide breakdown in COS-7 cells. Exposure to 10 microM ADP-beta-S blocked the increase in [Ca2+]i induced in the Ca(2+)-free medium by hypotonic stimulation. The results of summary points 4, 5, and 6 suggest that the increase in [Ca2+]i induced by hypotonic stimulation is due to Ca2+ release from inositol 1,4,5-trisphosphate (IP3)-sensitive internal stores. 7. The hypotonic stimulation-activated hydrolysis of phosphoinositides was decreased by pertussis toxin (PTX) in a dose-dependent manner. 8. These observations strongly suggest that hypotonic stimulation induced an increase in [Ca2+]i in Ca(2+)-free medium through activation of cascades using PTX-sensitive guanine nucleotide binding protein (G protein) and IP3.

MeSH Terms
Animals Caffeine/pharmacology Calcium/metabolism Cell Line Central Nervous System Stimulants/pharmacology Chlorocebus aethiops Culture Media Fluorescent Dyes Fura-2 GTP-Binding Proteins/metabolism Hypotonic Solutions Inositol 1,4,5-Trisphosphate/pharmacology Kidney/drug effects,metabolism Membrane Potentials/drug effects Patch-Clamp Techniques Pertussis Toxin Phosphatidylinositols/metabolism Virulence Factors, Bordetella/pharmacology
Chemicals
Central Nervous System Stimulants Culture Media Fluorescent Dyes Hypotonic Solutions Phosphatidylinositols Virulence Factors, Bordetella Caffeine Inositol 1,4,5-Trisphosphate Pertussis Toxin GTP-Binding Proteins Calcium Fura-2
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ishii T
Department of Physiology, Faculty of Medicine, Kyoto University, Japan.
Hashimoto T
Ohmori H
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Article Info
Journal
The Journal of physiology
Abbr.
J Physiol
ISSN
0022-3751
DOI
10.1113/jphysiol.1996.sp021389
Published
1996-06-01
Pages
371-84
Language
English
Region
England
NLM ID
0266262
PMCID
PMC1158923
Subset
IM
External Links
PubMed source DOI: 10.1113/jphysiol.1996.sp021389 PMC Full Text View journal details
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