Comparison of cytosolic Ca2+ and exocytosis responses from single rat and bovine chromaffin cells.
The relationship between cytosolic Ca2+ and catecholamine secretion during stimulus-secretion coupling has been examined at individual chromaffin cells isolated from the cow and rat. Vesicular catecholamine exocytosis was determined via amperometric measurements with carbon fibre microelectrodes and fura-2 was used for simultaneous fluorescent monitoring of cytosolic Ca2+ at the same cell. Individual secretory vesicles in cells from the two species were found to release similar amounts of catecholamine. In addition, the time courses for secretion from individual vesicles was similar with rat and bovine chromaffin cells. The total quantity of catecholamine released and the change in cytosolic Ca2+ were also similar in response to exposure to K+ (60 mM), 1,1-dimethyl-4-phenylpiperazinium (50 microM), and histamine (50 microM), although both responses were more prolonged following 1,1-dimethyl-4-phenylpiperazinium and histamine at bovine cells. Agents that mobilize intracellular Ca(2+)-stores such as methacholine, caffeine and bradykinin resulted in different cytosolic Ca2+ and exocytosis responses at the rat and bovine chromaffin cells. Results indicate a heightened Ca(2+)-store activity or a more filled state in chromaffin cells from the rat. The results of this study clearly show that single-cell techniques can be used to characterize stimulus-secretion coupling. The requirement for lower numbers of cells with these techniques means that chromaffin cells from rodents can be routinely employed. This can be advantageous to minimize biological variability which occurs with organs obtained from slaughter houses and enables the investigation of genetically-altered animals.
Finnegan
J M
JM
Department of Chemistry, University of North Carolina at Chapel Hill 27599-3290, USA.
Borges
R
R
Wightman
R M
RM
D000818:Animals; D001920:Bradykinin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002395:Catecholamines / Q000378:metabolism*; D002417:Cattle; D002838:Chromaffin System / Q000378:metabolism*; D004246:Dimethylphenylpiperazinium Iodide / Q000494:pharmacology; D005089:Exocytosis / Q000502:physiology*; D006632:Histamine / Q000494:pharmacology; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley
D002395:Catecholamines; D004246:Dimethylphenylpiperazinium Iodide; D006632:Histamine; D001920:Bradykinin; D002118:Calcium