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Titolo:
HEK-293 cells possess a carbachol- and thapsigargin-sensitive intracellular Ca2+ store that is responsive to stop-flow medium changes and insensitiveto caffeine and ryanodine
Autore:
Tong, JF; Du, GG; Chen, SRW; MacLennan, DH;
Indirizzi:
Univ Toronto, Banting & Best Dept Med Res, Toronto, ON M5G 1L6, Canada Univ Toronto Toronto ON Canada M5G 1L6 d Res, Toronto, ON M5G 1L6, Canada Univ Calgary, Dept Biochem & Mol Biol, Cardiovasc Res Grp, Calgary, AB T2N4N1, Canada Univ Calgary Calgary AB Canada T2N 4N1 es Grp, Calgary, AB T2N4N1, Canada
Titolo Testata:
BIOCHEMICAL JOURNAL
, volume: 343, anno: 1999,
parte:, 1
pagine: 39 - 44
SICI:
0264-6021(19991001)343:<39:HCPACA>2.0.ZU;2-E
Fonte:
ISI
Lingua:
ENG
Soggetto:
SINGLE-CHANNEL PROPERTIES; HEK293 CELLS; RECEPTOR; RELEASE; EXPRESSION; CALCIUM; KIDNEY; MUTATIONS; RETICULUM; TRANSPORT;
Keywords:
Ca2+ photometry and imaging; Ca2+ release channels; ryanodine receptors;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
22
Recensione:
Indirizzi per estratti:
Indirizzo: MacLennan, DH Univ Toronto, Banting & Best Dept Med Res, 112 Coll St, Toronto, ON M5G 1L6, Canada Univ Toronto 112 Coll St Toronto ON Canada M5G 1L6 6, Canada
Citazione:
J.F. Tong et al., "HEK-293 cells possess a carbachol- and thapsigargin-sensitive intracellular Ca2+ store that is responsive to stop-flow medium changes and insensitiveto caffeine and ryanodine", BIOCHEM J, 343, 1999, pp. 39-44

Abstract

Because HEK-293 cells are widely used for the functional expression of channels, exchangers and transporters involved in Ca2+ homoeostasis, the properties of intracellular Ca2+ stores and the methods used for measuring intracellular Ca2+ release in HEK-293 cells were evaluated. Ca2+ imaging was used to show caffeine-, carbachol- and thapsigargin-induced Ca2+ release in HEK-293 cells transfected with ryanodine receptor (RyR) cDNA, but only carbachol- and thapsigargin-induced Ca2+ release in untransfected HEK-293 cells. Intracellular Ca2+ release in untransfected HEK-293 cells was also observedif medium changes were performed by aspirating and replacing fresh medium (stop-flow), but not if medium changes were performed by a continuous over-flow procedure. Stop-flow medium-change-induced Ca2+ release in HEK-293 cells was independent of caffeine and ryanodine, demonstrating that it did notoccur through RyR channels. Consistent with these observations was the observation that the level of expression of endogenous RyR proteins was below the limits of detection by Western blotting or [H-3]ryanodine binding. Thusthe level of endogenous expression of RyR is so low in HEK-293 cells as toprovide a negligible background in relation to Functional analysis of recombinant RyR molecules. These results are inconsistent with those of Querfurth et al. [Querfurth, Haughey, Greenway, Yacono, Golan and Geiger (1998) Biochem. J. 334, 79-86], who reported higher levels of endogenous RyR expression in untransfected HEK-293 cells.

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Documento generato il 03/07/20 alle ore 16:08:54