Opioids and cytosolic calcium in rat anterior pituitary: dynorphin preparation showed LHRH-like action due to contamination.
The effect of dynorphin A-(1-13) (Dyn A-(1-13] and other opioids on the cytosolic free calcium concentration [(Ca2+]i) in rat anterior pituitary cells was examined using the fluorescent indicator fura-2. A commercial synthetic Dyn A-(1-13) preparation elevated [Ca2+]i. Results, which were obtained with receptor antagonists, and in LHRH receptor radioligand binding studies as well as by HPLC combined with LHRH radioimmunoassay, strongly suggest that this effect of the dynorphin preparation was due to contamination with a LHRH-like compound. Dyn A-(1-13), purified by HPLC, as well as Dyn A-(2-13), [Leu5]enkephalin, beta-endorphin, morphine, or U50,488H had no effect on [Ca2+]i. LHRH caused a rapid increase in [Ca2+]i by about 50 nM which was blocked by the LHRH antagonist, [D-pGlu1,D-Phe2,D-Trp3,6] LHRH.
Knepel
W
W
Department of Pharmacology, University of Freiburg im Breisgau, Federal Republic of Germany.
Schöfl
C
C
Wesemeyer
G
G
Götz
D M
DM
D000818:Animals; D001572:Benzofurans; D002118:Calcium / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004340:Drug Contamination; D004399:Dynorphins / Q000302:isolation & purification / Q000494:pharmacology* / Q000592:standards; D004723:Endorphins / Q000502:physiology*; D005456:Fluorescent Dyes; D016257:Fura-2; D007987:Gonadotropin-Releasing Hormone / Q000032:analysis / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D008297:Male; D009270:Naloxone / Q000494:pharmacology; D010446:Peptide Fragments / Q000302:isolation & purification / Q000494:pharmacology* / Q000592:standards; D010903:Pituitary Gland, Anterior / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D001572:Benzofurans; D004723:Endorphins; D005456:Fluorescent Dyes; D010446:Peptide Fragments; D007987:Gonadotropin-Releasing Hormone; D009270:Naloxone; C026843:dynorphin (1-13); D004399:Dynorphins; D002118:Calcium; D016257:Fura-2