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PMID: 9442108 Published · ppublish English Journal Article

Capacitative Ca2+ entry is closely linked to the filling state of internal Ca2+ stores: a study using simultaneous measurements of ICRAC and intraluminal [Ca2+].

The Journal of cell biology ·Vol. 140 ·No. 2 ·1998-01-26 ·Pages 325-34

Hofer AM, Fasolato C, Pozzan T

Abstract

ICRAC (the best characterized Ca2+ current activated by store depletion) was monitored concurrently for the first time with [Ca2+] changes in internal stores. To establish the quantitative and kinetic relationship between these two parameters, we have developed a novel means to clamp [Ca2+] within stores of intact cells at any level. The advantage of this approach, which is based on the membrane-permeant low-affinity Ca2+ chelator N,N,N',N'-tetrakis (2-pyridylmethyl)ethylene diamine (TPEN), is that [Ca2+] within the ER can be lowered and restored to its original level within 10-15 s without modifications of Ca2+ pumps or release channels. Using these new tools, we demonstrate here that Ca2+ release-activated Ca2+ current (ICRAC) is activated (a) solely by reduction of free [Ca2+] within the ER and (b) by any measurable decrease in [Ca2+]ER. We also demonstrate that the intrinsic kinetics of inactivation are relatively slow and possibly dependent on soluble factors that are lost during the whole-cell recording.

MeSH Terms
Animals Calcium/metabolism Calcium Channels/metabolism Cell Line Chelating Agents/pharmacology Cytoplasm/metabolism Egtazic Acid/analogs & derivatives,pharmacology Ethylenediamines/pharmacology Fura-2/pharmacology Ionomycin/pharmacology Ionophores/pharmacology
Chemicals
Calcium Channels Chelating Agents Ethylenediamines Ionophores Egtazic Acid Ionomycin 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine Calcium Fura-2
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hofer A M
University of Padova, Department of Biomedical Sciences, CNR Center for Biomembranes, I-35121 Padova, Italy.
Fasolato C
Pozzan T
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.140.2.325
Published
1998-01-26
Pages
325-34
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2132570
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.140.2.325 PMC Full Text View journal details
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