Dynorphin-A and vasopressin release in the rat: a structure-activity study.

The effects on vasopressin (VP) release of three dynorphin-A fragments and two antidynorphin antisera were tested in vivo and in vitro. In vivo, the order of potency to inhibit VP release 30 min upon i.c.v. injection was: dynorphin-A-(1-17) > dynorphin-A-(1-13) > dynorphin-A-(1-8). l.c.v. co-administration of 10 nmoles of the specific endopeptidase-inhibitor cFPAAF-pAB and dynorphin-A-(1-8) also suppressed VP secretion. Dynorphin-A-(1-17) antiserum enhanced VP release 20 and 60 min after i.c.v. injection. The antiserum that recognized dynorphin-A-(1-13) elevated VP plasma levels at 60 min post-injection. In vitro, dynorphin-A-(1-8) suppressed electrically evoked VP release from the isolated neural lobe. VP release was not affected by dynorphin-A-(1-13), dynorphin-A-(1-17), naloxone, or by the anti-dynorphin antisera. These data indicate that dynorphin-A-(1-17), rather than dynorphin-A-(1-8), plays a role in the centrally located control of neurohypophysial VP release, whereas dynorphin-A-(1-8) is involved in the control located in the posterior pituitary. The synthetic intermediate fragment dynorphin-A-(1-13) appears to affect VP release both centrally and peripherally.

The opioid receptor subtypes mu and kappa, but not delta, are involved in the control of the vasopressin and oxytocin release in the rat.

The effects of highly selective agonists and antagonists to the mu-, delta- and kappa-opioid receptor subtypes were studied on the vasopressin and oxytocin release in 24 h water-deprived male rats. The delta-agonist [D-Pen2,D-Pen5]enkephalin (dose range 0.01-5 mg/kg) did not affect plasma levels of either hormone 30 min after s.c. administration, whereas the mu-agonist DALDA (H-Tyr-D-Arg-Phe-Lys-NH2) over the same dose range strongly inhibited the release of both vasopressin and oxytocin, an effect that was maximal 30-60 min after s.c. injection. The same effect was found for s.c. administration of the kappa-agonist U-69,593. Intracerebroventricular (i.c.v.) administration of DALDA (0.5 and 5 micrograms/kg) but not U-69,593 suppressed both plasma hormone levels 30 min after injection. Also the effects of selective antagonists were tested over the s.c. dose range of 0.01-1 mg/kg. Whereas both the kappa-selective antagonist nor-binaltorphimine and the relatively mu-selective antagonist naloxone elevated oxytocin plasma levels (peak at 15 and 30 min after injection, respectively), the delta-selective antagonist naltrindole was without any effect. Nor-binaltorphimine, naloxone, and naltrindole did not affect vasopressin release. When the antagonists were administered i.c.v. (dose range 2.5-25 micrograms/kg), only the kappa-antagonist nor-binaltorphimine enhanced oxytocin and vasopressin release 30 min after injection. In conclusion, both mu- and kappa-opioid receptors are involved in the regulation of the secretion of vasopressin and oxytocin from the rat neural lobe; in contrast, delta-opioid receptors do not play a role.(ABSTRACT TRUNCATED AT 250 WORDS)

Pharmacological assessment of the site of action of opioids on the release of vasopressin and oxytocin in the rat.

Naloxone and its congener, methyl naloxone, were given subcutaneously (s.c.) or centrally (i.c.v.) to 24-h water-deprived male rats 30 min prior to decapitation and the effect on plasma levels of vasopressin (VP) and oxytocin (OT) was studied. The potency of s.c. applied methyl naloxone to increase plasma OT levels did not differ from that of naloxone. Injected i.c.v., neither methyl naloxone nor naloxone had a clear effect and they antagonized i.c.v. co-administered dynorphin A-(1-13) equipotently. Methyl naloxone or naloxone, s.c., antagonized the inhibitory action of simultaneous dynorphin A-(1-13) and beta-endorphin-(1-31) given i.c.v., although higher doses of methyl naloxone were required. The data indicate that the sites of inhibition of neurohypophysial hormone release due to beta-endorphin-(1-31) are more likely to be located mostly within the blood-brain barrier, to which methyl naloxone has less ready access, than are the sites of inhibition due to dynorphin A-(1-13).

Solid-phase extraction of plasma vasopressin: evaluation, validation and application.

A new solid-phase extraction method using octyl-silica columns to extract vasopressin-like immunoreactivity from plasma has been developed. The extraction was followed by a radioimmunoassay on the vacuum-dried extracts, which were reconstituted in assay buffer. The total recovery of synthetic vasopressin was ca. 100%. Based on co-elution with synthetic vasopressin after separation by reversed-phase high-performance liquid chromatography of plasma extracts from normal Wistar and Brattleboro rats, and the cross-reactivity of the antiserum used in the radioimmunoassay system, the extracted material was found to be indistinguishable from authentic vasopressin. Unknown experimental samples were interpolated on a standard curve established in "zero" plasma (plasma derived from rats subjected to waterload) spiked with known amounts of synthetic vasopressin, and not on a standard curve established in assay buffer. The limit of detection was 1 fmol of vasopressin equivalent per millilitre. The intra- and inter-assay coefficients of variance were 10-16% and 16%, respectively. The procedure reliably showed that osmotic challenge and 24-h dehydration increased, whereas ethanol ingestion decreased vasopressin-like immunoreactivity plasma levels in the rat, compared with normally hydrated controls.