Time-course of capsaicin-evoked release of calcitonin gene-related peptide from rat spinal cord in vitro. Effect of concentration and modulation by Ruthenium Red.
The capsaicin-evoked release of calcitonin gene-related peptide (CGRP) from rat superfused dorsal spinal cord slices was investigated during sustained capsaicin exposure thought to represent equilibrium conditions. The dose-effect relationship for total peptide release evoked by single capsaicin doses (26 min exposure) was very steep with a threshold at 0.06 microM and a maximum at 0.3 microM capsaicin. With concentrations of capsaicin within this range the slow decline of the peptide release in the presence of capsaicin was not a consequence of exhaustion of an available peptide pool nor of neuronal impairment because potassium depolarization was still able to release CGRP. In contrast, with concentrations of capsaicin at 1.5 microM and above, there was a much faster decay of the release after the peak, most probably due to a loss of the secretion capacity caused by neuronal inactivation. When cumulative dose regimens for capsaicin were employed, release of CGRP could be stimulated only up to a dose of 1-1.5 microM capsaicin; further increase in capsaicin concentration was ineffective. This was also most probably due to a loss of the secretion capacity caused by neuronal inactivation and not caused by depletion of a releaseable peptide pool. Release of CGRP evoked by capsaicin concentrations in the range of 0.1-0.3 microM in either dosage protocol was reduced in the presence of Ruthenium Red (RR, 2.5 microM). RR did not reduce neuropeptide release evoked by capsaicin concentrations at or above 1-1.5 microM, nor did it affect the inactivation of the release process at such high capsaicin concentrations. The results demonstrate that, upon sustained exposure to capsaicin, different ranges of concentration can be established at which either only stimulatory or a mixture of stimulatory and inhibitory effects determine the amount of neuropeptides released.
Donnerer
J
J
Department of Experimental and Clinical Pharmacology, University of Graz, Austria.
Schuligoi
R
R
Amann
R
R
D000818:Animals; D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism*; D002211:Capsaicin / Q000494:pharmacology*; D002851:Chromatography, High Pressure Liquid; D066298:In Vitro Techniques; D007700:Kinetics; D051381:Rats; D011919:Rats, Inbred Strains; D012430:Ruthenium Red / Q000494:pharmacology*; D013116:Spinal Cord / Q000378:metabolism*
D012430:Ruthenium Red; D015740:Calcitonin Gene-Related Peptide; D002211:Capsaicin