(+)-isradipine but not (-)-Bay-K-8644 exhibits voltage-dependent effects on cat adrenal catecholamine release.
1. Catecholamine release from cat adrenal glands perfused at a high rate (4 ml min-1) at 37 degrees C with polarizing (1.2 or 5.9 mM K+) or depolarizing (17.7, 35, 59 or 118 mM K+) solutions, was triggered by 5 or 10 s pulses of Ca2+ (0.5 or 2.5 mM) in the presence of various concentrations of K+. 2. In polarized glands, secretion was greater the higher the K+ concentration present during the secretory K+/Ca2+ test pulse. Thus, in 17.7 mM K+, catecholamine released was 162 +/- 27 ng per pulse, while in 118 mM K+ secretion rose to 1839 +/- 98 ng per pulse. In depolarized glands, secretion reached a peak of around 1000 ng per pulse in 35-59 mM K+; in 118 mM K+, secretion did not increase further, suggesting that voltage changes are implicated in the control of the secretory process. 3. Blockade of secretion by increased concentrations of (+)-isradipine was much more manifest in polarized glands. The higher the degree of depolarization was (35, 59 or 118 mM K+), the lower the IC50 s were. So, the ratios between the IC50 s in polarized and depolarized glands rose from 3.92 in 35 mM K+ to 26.7 in 118 mM K+. 4. In contrast, the Ca2+ channel activator (-)-Bay K 8644 potentiated catecholamine release evoked by K+/Ca2+ pulses equally well in polarized or depolarized glands. The ratios between EC50 s in polarized or depolarized glands were, respectively, 0.30, 0.59 and 0.69 for 17.7, 35 and 118 mM K+. 5. In simultaneous experiments, the two enantiomers of Bay K 8644 exhibited opposite effects on secretion. (+)-Bay K 8644 (a Ca21 channel blocker) inhibited secretion better in depolarized than in polarized glands, whilst (-)-Bay K 8644 potentiated secretion in a voltage-independent manner. 6. Potentiation of secretion by (-)-Bay K 8644 was concentration-dependent from 10-8 to 10-6M. At 10- 5M, such potentiation largely disappeared in both polarized and depolarized glands. However, this dual effect of (-)Bay K 8644 was better seen in depolarizing conditions, suggesting that using the same enantiomer, the voltage-dependence is only seen when blockade of secretion dominates. 7. In the presence of increasing concentrations of (-)Bay K 8644 (3 x 10-9, 3 x 10-8 and 3 x 10-7M), the concentration-response curves for (+)isradipine to inhibit secretion were displaced to the right. However, a Schild plot of (dose ratio - 1) against (-)-Bay K 8644 concentrations gave a slope of 0.6, suggesting that the interactions between (+)-isradipine and (-)Bay K 8644 were non-competitive in nature. The pA2 for (-)-Bay K 8644 was 9.13. 8. Overall, the results suggest that potentiation of secretion by (-)Bay K 8644 (a voltage-independent phenomenon), and blockade by (+)-isradipine or (+-Bay K 8644 (a voltage-dependent phenomenon) might be exerted through binding of the dihydropyridines activators and blockers to separate sites on chromaffin cell L-type Ca2 + channels.
López
M G
MG
Departamento de Farmacología, Facultad de Medicina, Universidad Autónoma de Madrid, Spain.
Michelena
P
P
Gandía
L
L
García
A G
AG
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000737:chemistry / Q000494:pharmacology; D000311:Adrenal Glands / Q000187:drug effects* / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002395:Catecholamines / Q000502:physiology*; D002415:Cats; D004563:Electrochemistry; D005260:Female; D066298:In Vitro Techniques; D017275:Isradipine; D008297:Male; D011188:Potassium / Q000494:pharmacology; D011725:Pyridines / Q000737:chemistry / Q000494:pharmacology*; D013237:Stereoisomerism
D002121:Calcium Channel Blockers; D002395:Catecholamines; D011725:Pyridines; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D011188:Potassium; D002118:Calcium; D017275:Isradipine