The effect of thrombin and serine proteases on intracellular Ca2+ in rat aortic smooth muscle cells.
Cultures of vascular smooth muscle cells (VSMC) are commonly used to study the events and defects found in hypertension and atherosclerosis. In particular Ca2+ homeostasis in cellular signalling has been the focus of extensive research. Since trypsin has been shown to mobilise Ca2+ in some cell types, we have investigated its effect on various aspects of Ca2+ homeostasis in rat aortic smooth muscle cells (RASMC). The effects of trypsin, alpha-chymotrypsin and elastase (other serine proteases) on intracellular Ca2+ in cultured aortic cells isolated from Wistar rats have been investigated. Trypsin (24 micrograms/ml) elicits intracellular Ca2+ mobilisation, after which cells become nonresponsive to thrombin Ca2+ mobilisation but retain responsiveness to Angiotensin II (AII). alpha-Chymotrypsin (24 micrograms/m) inhibits the thrombin Ca2+ mobilising response, without itself initiating a Ca2+ transient or affecting AII Ca2+ mobilisation. Elastase (24 micrograms/ml) was not effective in mobilising intracellular Ca2+ or inhibiting the thrombin response. We have also observed diminished thrombin Ca2+ mobilisation responses between cells in suspension and cell monolayers, which appeared to be unrelated to proteolysis but due to morphological changes of the cells. Our results suggest that trypsin acts on the thrombin receptor via a specific proteolysis mechanism to mobilise intracellular Ca2+ ([Ca2+]i) in RASMC. The amount of Ca2+ released by thrombin or trypsin is dependent on the morphology of the cell and the state of the tethered ligand of the thrombin receptor exposed by the protease.
Kable
E P
EP
Department of Pharmacy, Sydney University, NSW, Australia.
Monteith
G R
GR
Roufogalis
B D
BD
D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D007424:Intracellular Fluid / Q000187:drug effects / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism*; D051381:Rats; D012697:Serine Endopeptidases / Q000494:pharmacology*; D013917:Thrombin / Q000494:pharmacology*
D000804:Angiotensin II; D012697:Serine Endopeptidases; D013917:Thrombin; D002118:Calcium