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Characterization of calcium signaling pathways in human preadipocytes
Author(s) -
Hu Rui,
He Mulan,
Hu Hao,
Yuan BingXiang,
Zang WeiJin,
Lau ChuPak,
Tse HungFat,
Li GuiRong
Publication year - 2009
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.21823
Subject(s) - thapsigargin , serca , cyclopiazonic acid , ryanodine receptor , trpc , phospholipase c , inositol trisphosphate receptor , endoplasmic reticulum , microbiology and biotechnology , chemistry , orai1 , signal transduction , calcium signaling , inositol , stim1 , calcium , medicine , endocrinology , biology , receptor , biochemistry , transient receptor potential channel , atpase , organic chemistry , enzyme
Intracellular free Ca 2+ (Ca i 2+ ) is an important regulator of many cellular activities; however, Ca 2+ signaling is not well studied in human preadipocytes. The purpose of the present study was to characterize Ca 2+ signal pathways using a confocal scanning technique and RT‐PCR. It was found that spontaneous Ca i 2+oscillations were observed in 12.1% preadipocytes, and number of cells with Ca 2+ oscillations was increased to 47.9% by 1% fetal bovine serum. Ca i 2+oscillations were dependent on Ca 2+ entry mainly via stored‐operated Ca 2+ (SOC) entry. They were suppressed by the SOC entry channel blocker La 3+ , the phospholipase C (PLC) inhibitor U73122, the inositol trisphosphate receptor (IP3R) blocker 2‐amino‐ethoxydiphenyl borate, or the sarcoplasmic/endoplasmic reticulum Ca 2+ pump (SERCA) inhibitors thapsigargin and cyclopiazonic acid, but not by ryanodine. The IP3R activator thimerosal increased Ca i 2+oscillations. In addition, the plasma membrane Ca 2+ pump (PMCA) inhibitor carboxyeosin and Na + –Ca 2+ exchanger (NCX) inhibitor Ni 2+ both suppressed Ca 2+ oscillations. RT‐PCR revealed that the mRNAs for IP3R1‐3, SERCA1,2, NCX3 and PMCA1,3,4, Ca V 1.2, and TRPC1,4,6, STIM1 and Orai1 (for SOC entry channels) were significant in human preadipocytes. The present study demonstrates that multiple Ca 2+ signal pathways are present in human preadipocytes, and provides a basis for investigating how Ca 2+ signals regulate biological and physiological activities of human preadipocytes. J. Cell. Physiol. 220: 765–770, 2009. © 2009 Wiley‐Liss, Inc.