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C. Meyer, M. J. Freund-Mercier, Y. Guerné, and Ph. Richard

ABSTRACT

Plasma concentrations of oxytocin and vasopressin were measured in relationship to oxytocin cell firing during suckling in urethane-anaesthetized rats.

Preliminary experiments showed that plasma concentrations of oxytocin and vasopressin, which were increased immediately after anaesthesia, reverted to basal concentrations 3 h later. Moreover, it was found that exogenous oxytocin had entirely disappeared 5 min after i.v. bolus injections of known doses of oxytocin.

Suckling did not modify the basal plasma concentration of oxytocin (14·6 ± 2·9 compared with 14·±61·5 pmol/l before suckling) except during a brief period immediately after neurosecretory bursts on oxytocin cells (37·8 ± 5·2 pmol/l; P < 0·001, n = 11). The plasma concentration of oxytocin did not differ significantly from the basal concentration 1·5 min later. The plasma concentration of vasopressin never varied.

After two neurosecretory bursts of similar amplitude (total number of spikes during the burst) recorded on the same oxytocin cell, the variations in plasma concentration of oxytocin were also similar. When, for a given cell, the amplitude of neurosecretory bursts increased or decreased, the amount of oxytocin released changed in the same way.

These data demonstrate (1) that suckling induces pulsatile release of oxytocin without vasopressin, and (2) a direct relationship between the amounts of oxytocin released and the amplitude of oxytocin cell neurosecretory bursts which argue in favour of simultaneous increases or decreases in the neurosecretory burst amplitudes on all oxytocin cells.

J. Endocr. (1987) 114, 263–270

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F. Moos, M. J. Freund-Mercier, Y. Guerné, J. M. Guerné, M. E. Stoeckel, and Ph. Richard

ABSTRACT

The release of endogenous oxytocin and vasopressin by rat paraventricular and supraoptic nuclei in vitro during a 10-min period, 30 min after beginning the incubation, was measured radioimmunologically. Mean basal hormone release per 10 min and per pair of nuclei was: 128·4 ± 12·4 (s.e.m.) pg vasopressin (n = 15) and 39·0 ± 3·0 pg oxytocin (n = 66) for supraoptic nuclei from male rats; 273·9 ± 42·6 pg vasopressin (n = 11) and 34·2 ± 3·5 pg oxytocin (n = 15) for supraoptic nuclei from lactating rats; 70·0 ± 8·6 pg vasopressin (n = 52) and 21·8 ± 1·3 pg oxytocin (n = 68) for paraventricular nuclei from male rats; 59·1 ± 8·6 pg vasopressin (n = 10) and 27·0 ± 4·6 pg oxytocin (n = 16) for paraventricular nuclei from lactating rats.

In male and lactating rats, both nuclei contained and released more vasopressin than oxytocin. For oxytocin alone, the paraventricular nucleus of male rats contained and released significantly less hormone than the supraoptic nucleus. This difference was not apparent in lactating rats. For vasopressin alone, the paraventricular nucleus contained and released significantly less hormone than the supraoptic nucleus in both male and lactating rats. When the hormone released was calculated as a percentage of the total tissue content the release was about 0·9% for oxytocin from both nuclei in male and lactating rats and also for vasopressin in lactating rats, but was only about 0·5% for vasopressin from both nuclei in male rats.

The influence of oxytocin and analogues of oxytocin (including one antagonist) upon the release of oxytocin and vasopressin was studied. Adding oxytocin to the incubation medium (0·4–4 nmol/l solution) induced a dose-dependent rise in oxytocin release from both nuclei of male or lactating rats. A 4 nmol/l solution of isotocin had a similar effect to a 0·4 nmol/l solution of oxytocin, but arginine-vasopressin never affected basal release of oxytocin. In no case was vasopressin release modified.

An oxytocin antagonist (1 μmol/l solution) significantly reduced basal oxytocin release and blocked the stimulatory effect normally induced by exogenous oxytocin, as did gallopamil hydrochloride (D600, 10 μmol/l solution), a Ca2+ channel blocker, or incubation in a Ca2+-free medium.

These findings are discussed in relation to the literature on the central effects of neurohypophysial peptides. It may be concluded that the regulatory role of endogenous oxytocin in the hypothalamus on the milk-ejection reflex could result from its local release in the extracellular spaces of magnocellular nuclei.

J. Endocr. (1984) 102, 63–72

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M. E. Stoeckel, M. J. Freund-Mercier, J. M. Palacios, Ph. Richard, and A. Porte

ABSTRACT

The distribution of [3H]vasopressin- and [3H]oxytocin-binding sites was examined, using an autoradiographical technique, in the kidney of Long–Evans and Brattleboro rats. Two types of binding sites with affinities in the nanomolar range were detected: one, located on glomeruli, bound both vasopressin and oxytocin; the other, on collecting ducts, bound vasopressin selectively. In the presence of 10 μmol oxytocin/1, [3H]vasopressin labelling was abolished in glomeruli, but only reduced in collecting ducts; [3H]oxytocin labelling was completely abolished by 10 μmol vasopressin/1. These observations are discussed in relation to known effects of neurohypophysial hormones on renal physiology.

J. Endocr. (1987) 113, 179–182

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M P Arpin-Bott, E Waltisperger, M J Freund-Mercier, and M E Stoeckel

Abstract

The localization of oxytocin (OT)-binding sites in the developing rat kidney and their pharmacological characterization were investigated by means of autoradiographic techniques. The cellular localization was studied by application of the histoautoradiographic technique to (1) frozen sections and semithin sections from kidney slices incubated in vitro in the presence of a 125I-labelled OT antagonist and (2) frozen and semithin sections from kidneys after in vivo systemic infusion of the radioligand. Pharmacological characteristics were determined in competition experiments by using quantitative film autoradiography.

Specific OT-binding sites were first detected at embryonic day 17 (E17) in the cortex. At early stages up to postnatal days (PN30), the cortical OT-binding sites were highly concentrated on the juxta- and paraglomerular portion of the distal tubule; in the adult they were restricted to the macula densa. In the medulla, OT-binding sites were first detected at E19 when this region is forming; they were localized on the thin limb of Henle's loop. These data obtained by in vitro binding were confirmed by in vivo binding at PN30 which showed, in addition, the presence in one rat of OT-binding sites in the inner stripe of the outer medulla.

At all stages examined (PN15 to PN90), cortical OT-binding sites had a higher selectivity for OT versus vasopressin (IC50=0·78 ± 0·04 nm and 8 ± 0·5 nm respectively at PN90) than medullary sites (IC50= 1·9 ± 0·27 nm and 2±1·13 nm respectively at PN90). These data suggest that the OT-binding sites of the macula densa and thin Henle's loop, detected in the rat kidney, represent two subtypes of OT receptors which could mediate distinct effects of OT on kidney function.

Journal of Endocrinology (1997) 153, 49–59