Thapsigargin and EGTA inhibit endothelin-1-induced glucose transport.

We have previously demonstrated that ET-1 may enhance glucose transport in 3T3-L1 adipocytes, secondarily to its stimulatory effect on GLUT1 gene expression by a mitogen-activated protein kinase (MAPK)-dependent pathway. In the present study, we further tested the involvement of Ca(2+) in glucose uptake in response to ET-1. Among a variety of Ca(2+)-related agents tested, EGTA and thapsigargin were found to suppress both the glucose uptake and intracellular Ca(2+) mobilization induced by ET-1, as determined by Fura-2 analysis. However, a phospholipase C inhibitor, U73122, also eliminated the intracellular calcium mobilization induced by ET-1, but had no effect on ET-1-stimulated glucose uptake. The finding that neither EGTA nor thapsigargin had any influence on ET-1-induced MAPK activation implies that some mechanism downstream of MAPK activation is involved. Further investigation showed that both agents exerted global inhibitory effects on protein and RNA syntheses. Since both thapsigargin and EGTA may deplete endoplasmic reticulum (ER) Ca(2+) stores, our results suggest that (1) ET-1-induced glucose transport is independent of ET-1's effect on Ca(2+) mobilization and (2) depletion of ER Ca(2+) stores per se may interfere with ET-1's effect on GLUT1 expression.