open access

Vol 67, No 5 (2016)
Original papers
Published online: 2016-01-27
Submitted: 2015-11-18
Accepted: 2016-01-12
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The role of melatonin membrane receptors in melatonin-dependent oxytocin secretion from the rat hypothalamo–neurohypophysial system — an in vitro and in vivo approach

Marlena Juszczak, Monika Wolak, Ewa Bojanowska, Lucyna Piera, Magdalena Roszczyk
DOI: 10.5603/EP.a2016.0035
·
Endokrynologia Polska 2016;67(5):507-514.

open access

Vol 67, No 5 (2016)
Original papers
Published online: 2016-01-27
Submitted: 2015-11-18
Accepted: 2016-01-12

Abstract

Introduction: Melatonin exerts its biological role acting mainly via G protein-coupled membrane MT1 and MT2 receptors. To determine whether a response of oxytocinergic neurons to different concentrations of melatonin is mediated through membrane MT1 and/or MT2 receptors, the effect of melatonin receptors antagonists, i.e. luzindole (a non-selective antagonist of both MT1 and MT2 receptors) and 4-phenyl-2-propionamidotetralin (4-P-PDOT — a selective antagonist of MT2 receptor), on melatonin-dependent oxytocin (OT) secretion from the rat hypothalamo-neurohypophysial (H-N) system, has been studied both in vitro and in vivo.

Material and methods: For in vitro experiment, male rats served as donors of the H-N explants, which were placed in 1 ml of normal Krebs-Ringer fluid (nKRF) heated to 37oC. The H-N explants were incubated successively in nKRF {fluid B1} and incubation fluid as B1 enriched with appropriate concentration of melatonin, i.e. 10–9 M, 10–7 M, or 10–3 M and luzindole or 4-P-PDOT, or their vehicles (0.1% ethanol or DMSO) {fluid B2}. After 20 minutes of incubation in fluid B1 and then B2, the media were collected and immediately frozen before OT estimation by the RIA. The OT secretion was determined by using the B2/B1 ratio for each H-N explant. During in vivo experiment, rats were given an intracerebroventricular (i.c.v.) infusion of 5 mL luzindole or 4-P-PDOT, or their solvent (0.1% DMSO) and 10 minutes later the next i.c.v. infusion of 5 mL of either melatonin solution (10–7 M) or its vehicle (0.1 % ethanol in 0.9% sodium chloride).

Results: Melatonin at a concentration of 10-3 M significantly stimulated, while at a concentration of 10–9 M had no effect on, oxytocin secretion from the rat H-N system in vitro, also when luzindole or 4-P-PDOT was present in a medium. On the other hand, melatonin at a concentration of 10–7 M diminished this neurohormone output from an isolated H-N system and into the blood. Luzindole significantly suppressed such melatonin action, while 4-P-PDOT did not change the inhibitory influence of 10–7 M melatonin on oxytocin release, both in vitro and in vivo.

Conclusions: The present study demonstrates that an inhibitory effect of 10–7 M melatonin on oxytocin secretion from the rat H-N system is mediated through a subtype MT1 membrane receptor and its action is independent of subtype MT2 receptor. However, for the stimulatory effect of pharmacological concentration (10–3 M) of the pineal hormone on oxytocin release, probably mechanisms other than membrane MT1/MT2 receptor(s)-dependent are involved. (Endokrynol Pol 2016; 67 (5): 507–514)

Abstract

Introduction: Melatonin exerts its biological role acting mainly via G protein-coupled membrane MT1 and MT2 receptors. To determine whether a response of oxytocinergic neurons to different concentrations of melatonin is mediated through membrane MT1 and/or MT2 receptors, the effect of melatonin receptors antagonists, i.e. luzindole (a non-selective antagonist of both MT1 and MT2 receptors) and 4-phenyl-2-propionamidotetralin (4-P-PDOT — a selective antagonist of MT2 receptor), on melatonin-dependent oxytocin (OT) secretion from the rat hypothalamo-neurohypophysial (H-N) system, has been studied both in vitro and in vivo.

Material and methods: For in vitro experiment, male rats served as donors of the H-N explants, which were placed in 1 ml of normal Krebs-Ringer fluid (nKRF) heated to 37oC. The H-N explants were incubated successively in nKRF {fluid B1} and incubation fluid as B1 enriched with appropriate concentration of melatonin, i.e. 10–9 M, 10–7 M, or 10–3 M and luzindole or 4-P-PDOT, or their vehicles (0.1% ethanol or DMSO) {fluid B2}. After 20 minutes of incubation in fluid B1 and then B2, the media were collected and immediately frozen before OT estimation by the RIA. The OT secretion was determined by using the B2/B1 ratio for each H-N explant. During in vivo experiment, rats were given an intracerebroventricular (i.c.v.) infusion of 5 mL luzindole or 4-P-PDOT, or their solvent (0.1% DMSO) and 10 minutes later the next i.c.v. infusion of 5 mL of either melatonin solution (10–7 M) or its vehicle (0.1 % ethanol in 0.9% sodium chloride).

Results: Melatonin at a concentration of 10-3 M significantly stimulated, while at a concentration of 10–9 M had no effect on, oxytocin secretion from the rat H-N system in vitro, also when luzindole or 4-P-PDOT was present in a medium. On the other hand, melatonin at a concentration of 10–7 M diminished this neurohormone output from an isolated H-N system and into the blood. Luzindole significantly suppressed such melatonin action, while 4-P-PDOT did not change the inhibitory influence of 10–7 M melatonin on oxytocin release, both in vitro and in vivo.

Conclusions: The present study demonstrates that an inhibitory effect of 10–7 M melatonin on oxytocin secretion from the rat H-N system is mediated through a subtype MT1 membrane receptor and its action is independent of subtype MT2 receptor. However, for the stimulatory effect of pharmacological concentration (10–3 M) of the pineal hormone on oxytocin release, probably mechanisms other than membrane MT1/MT2 receptor(s)-dependent are involved. (Endokrynol Pol 2016; 67 (5): 507–514)

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Keywords

oxytocin; melatonin; luzindole; 4-P-PDOT; melatonin receptors

About this article
Title

The role of melatonin membrane receptors in melatonin-dependent oxytocin secretion from the rat hypothalamo–neurohypophysial system — an in vitro and in vivo approach

Journal

Endokrynologia Polska

Issue

Vol 67, No 5 (2016)

Pages

507-514

Published online

2016-01-27

DOI

10.5603/EP.a2016.0035

Bibliographic record

Endokrynologia Polska 2016;67(5):507-514.

Keywords

oxytocin
melatonin
luzindole
4-P-PDOT
melatonin receptors

Authors

Marlena Juszczak
Monika Wolak
Ewa Bojanowska
Lucyna Piera
Magdalena Roszczyk

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