Studies on the vasoconstrictor action of melatonin and putative melatonin receptor ligands in the tail artery of juvenile Wistar rats
Authors: Ting K. N. , Dunn W. R. , Davies D. J. , Sugden D. , Delagrange P. , Guardiola‐Lemaître B. , Scalbert E. , Wilson V. G. 
Published: January 2005

In this study we compared the vasoconstrictor activity of melatonin in rat isolated tail artery using two different recording systems, the Halpern pressure myograph and the Halpern‐Mulvany wire myograph, with the view to determining a reliable method for obtaining pharmacological data on vascular melatonin receptors. In addition, we characterized the melatonin receptor in this preparation, using analogues of melatonin, and examined the activity of various naphthalenic derivatives with biological activity in non‐vascular models of melatonin receptors.

Using the Halpern pressure myograph, cumulative addition of melatonin (0.1 nM to 1 μM) produced direct vasoconstriction (19.3±6.4% reduction in lumen diameter, n=5) in five of 11 pressurized segments, with pEC50 of 9.14±0.17. Similarly, non‐cumulative application of melatonin caused vasoconstriction (19.7±4.6% reduction in lumen diameter, n=7) in seven of 20 preparations examined with pEC50 of 8.74±0.26. The selective alpha2‐adrenoceptor agonist, UK‐14304 (5‐bromo‐6‐[2‐imidazolin‐2‐ylamino]‐quinoxaline bitartrate), produced vasoconstriction in all ‘melatonin‐insensitive’ preparations.

Melatonin (0.1 nM to 1 μM) failed to elicit isometric contractions of tail artery segments in the Halpern wire myograph, but produced concentration‐dependent potentiation of electrically‐evoked, isometric contractions (maximum effect of 150–200% enhancement) when applied either non‐cumulatively (seven of seven preparations) or cumulatively (four of seven preparations). The pEC50 value of melatonin (non‐cumulative) was 8.50±0.10 (n=7) which was not different from that obtained in the pressure myograph. All further experiments were conducted using a non‐cumulative protocol against electrically‐evoked, isometric contractions.

Based on the pEC50 values for the melatonin analogues examined, the pharmacological profile for the enhancement of electrically‐evoked contractions was 2‐iodomelatonin>6‐chloromelatonin(−)‐AMMTC□thinsp;5 S21634melatoninS20098>S20242S20304>6‐ hydroxymelatonin>S20932> (+) ‐AMMTC>N‐acetyl‐5‐HT. Our data suggests the vascular receptor belongs to the MEL1‐like subtype. All the indole‐based analogues of melatonin, 2‐iodomelatonin, (−)‐AMMTC, (+)‐AMMTC, S20932, 6‐chloromelatonin, 6‐hydroxymelatonin and N‐acetyl‐5‐HT, behaved as full agonists. All the naphthalenic derivatives examined, S21634, S20098, S20242 and S20304 behaved as partial agonists relative to melatonin.

The naphthalenic‐based antagonists, S20928 and S20929, did not modify electrically‐evoked, isometric contractions of the tail artery, but produced a parallel, rightward displacement of the melatonin concentration‐response curve. Based upon the effect of 1 μM S20928 and S20929, the estimated pKB values for these antagonists were 7.18±0.25 (n=4) and 7.17±0.25 (n=5), respectively.

We demonstrated that enhancement of electrically‐evoked, isometric contractions of the rat isolated tail artery (using the Halpern‐Mulvany wire myograph) is a simple and reproducible model for assessing the activity of putative agonists, partial agonists and antagonists at vascular melatonin receptors. Pharmacological characterization of the receptor suggests the presence of a MEL1‐like subtype.