A sexual dimorphism in hypothalamic arginine vasotocin (AVT) gene expression and AVT plasma levels in the Japanese quail (Coturnix coturnix japonica) in response to water deprivation.

To examine a possible sexual dimorphism in the osmotic control of arginine vasotocin (AVT) release in birds, age-matched male and female Japanese quail were subjected to water deprivation. The observed increased plasma osmolalities were accompanied by increased plasma AVT levels. Plasma mesotocin levels did not change with water deprivation. The sensitivity of the osmotic control of AVT release as determined by the slope of the relationship between plasma AVT levels and plasma osmolalities was significantly (P < 0.05) higher in males than in females. By Northern blot analysis, levels of hypothalamic AVT gene transcripts were increased 2.3 +/- 0.14- and 3.5 +/- 0.13-fold in water-deprived male and female Japanese quail, respectively, compared to normally hydrated birds. Our data suggest gender-related differences in the osmotic control of AVT release and in hypothalamic AVT gene expression in the Japanese quail.

c-fos expression in the forebrain and brainstem of White Leghorn hens following osmotic and cardiovascular challenges.

In chickens, hyperosmolality and hemorrhage increase hypothalamic vasotocin (AVT) gene expression and stimulate the secretion of AVT from the posterior pituitary gland. In this study, c-fos expression was used to identify areas in the forebrain and brainstem of the domestic chicken that are activated following acute osmotic stress and hemorrhage-induced hypotension. Conscious hens were osmotically stimulated by administering a single intraperitoneal injection of 3 M NaCl (5 ml/kg). Urethane-anesthetized hens were bled to a mean systemic arterial pressure of 80-90 mm Hg and maintained at this blood pressure for 1 h with additional bleedings as required. In both studies, the expression of c-fos was determined in control and experimental birds by using Northern blot analysis and in situ hybridization analysis. Osmotic stress and hemorrhage-induced hypotension increased c-fos expression in the same brain regions. Prominent structures in the forebrain that expressed c-fos mRNA following acute osmotic stress and hemorrhage-induced hypotension included the supraoptic nucleus and paraventricular nucleus and nuclei within the hypothalamus that are anterior and ventral to the third ventricle. In the chicken, this region includes the organum subseptale, the o. vasculosum laminae terminalis, and the nucleus septalis medialis. In the brainstem, following either injection of 3 M NaCl or hemorrhage-induced hypotension, increased c-fos expression was observed in the nucleus of the solitary tract, parabrachial nucleus, area postrema, and locus ceruleus. Thus, the chicken central nervous system appears to use shared neuronal circuitry to stimulate hypothalamic AVT release in response to disturbances in body fluid composition and decreases in either systemic blood pressure or volume.

Plasma arginine-vasotocin and hydroosmotic status of the terrestrial pit viper Bothrops jararaca.

Peaks corresponding to arg-vasotocin obtained by HPLC from Sep-Pak C18 column extracts of Bothrops jararaca plasma were identified by radioimmunoassay and amino acid analysis. Plasma vasotocin and protein levels, osmolality, and L-cystine-di-beta-naphthylamidase were also compared in snakes under normal hydration conditions with or without chronic administration of vasotocin or in the presence of chronic hydroosmotic challenges. Sep-Pak C18 and radioimmunoassay were validated for the extraction and determination of this peptide, respectively (about 80% recovery). EDTA presented a protective action on this recovery compared to the use of heparin as anticoagulant for snake blood. A reduction of vasotocin content related to the time of incubation of this peptide added to snake plasma was detected by radioimmunoassay. Snake plasma activity also on L-cystine-di-beta-naphthylamide indicated that this vasotocin-destroying effect was due to hydrolysis by a cystine-aminopeptidase-like activity. Plasma levels of vasotocin revealed an unexpected dispersion and absent correlation with plasma levels of osmolality. Measurable vasotocin in a large number of snakes associated with lower levels of l-cystine-di-beta-naphthylamidase in acute than in chronic salt loading suggested the role of this enzyme activity in long-term regulation of the vasotocin system in this snake.

Arginine vasotocin gene expression and secretion during osmotic stimulation and hemorrhagic hypotension in hens.

In chickens, hyperosmolality stimulates the secretion of vasotocin (AVT) and up-regulates hypothalamic AVT gene expression. Hemorrhage, on the other hand, has not been considered an effective stimulus for AVT release in this species. The effects of acute osmotic stress and prolonged hemorrhagic hypotension on AVT gene expression and secretion were studied in White Leghorn hens. Conscious hens were osmotically stimulated by administering a single ip injection of 3 M NaCl (5 ml/kg). Urethane-anesthetized hens were bled to a mean arterial pressure of 80-90 mm Hg and the pressure was maintained within this range by additional bleeding. A total of about 30% of the estimated blood volume was removed. Both experiments were terminated after 1 hr of stimulation. Plasma AVT levels in the hyperosmotic and hypovolemic hens were 4- and 2-fold higher, respectively, compared to controls. Hypothalamic AVT mRNA levels, detected by Northern blot analysis, were 2.5- and 2-fold higher in the osmotically stimulated and hypotensive groups, respectively, compared to control groups. As determined by in situ hybridization, both osmotic stimulation and hypovolemia resulted in an increase in the number of AVT mRNA-containing neurons in the supra-optic and paraventricular nuclei. Our results indicate that, under the conditions used, hypotension and hyperosmolality are equally effective in stimulating AVT gene expression and secretion of AVT.

Arginine vasotocin gene expression in hypothalamic neurons is up-regulated in chickens drinking hypertonic saline: an in situ hybridization study.

Osmotic stress stimulates the release of the avian hypothalamic neuropeptide arginine vasotocin (AVT) into the peripheral circulation. We conducted the present study to investigate the effects of salt-loading on AVT secretion and AVT gene expression in specific hypothalamic nuclei in chickens. White Leghorn chickens were provided food ad lib and either water or 2% NaCl to drink. Both plasma osmolality and plasma AVT levels were significantly increased in chickens that drank 2% NaCl for either two or four days compared to that in chickens that drank water. Results from in situ hybridization analysis demonstrated an increase in the number of neurons expressing AVT mRNA in the supraoptic (SON) and paraventricular nuclei (PVN) in chickens provided 2% NaCl to drink compared to chickens that were provided water to drink. The number of grains per neuron increased in the PVN, but not in the SON of osmotically stimulated birds. Thus, increased osmolality resulting from ingestion of hypertonic saline is an effective stimulus to increase hypothalamic AVT mRNA content in chickens.

Changes in poly(A) tail length of arginine vasotocin messenger ribonucleic acid in the hypothalamus of water-deprived chickens.

The effects of water deprivation on the secretion of vasotocin (AVT) and expression of the AVT gene were studied in White Leghorn cockerels. Animals deprived of water for 4 days were compared with normally hydrated controls. Blood samples were obtained for measurements of plasma osmolality and AVT levels, and the hypothalamus was collected for extraction of total cellular RNA. A 519-bp AVT cDNA was prepared by reverse transcriptase-polymerase chain reaction and a 209-bp PstI/EcoRI restriction fragment from the 3' region of the fowl AVT cDNA was used as a probe for Northern blot analysis. Plasma osmolality and AVT levels in dehydrated birds were about 30 and 350% greater, respectively, than those in normally hydrated controls. The quantity of hypothalamic AVT mRNA was 2. 3-fold greater in water-deprived birds compared to controls. The size of the hypothalamic AVT transcript was about 100-bp longer in the water-deprived birds. As determined by RNase H treatment in the presence and absence of oligo(dT)12-18, the increase in mean size of the AVT mRNA in dehydrated animals was due to a longer poly(A) tract. Our results indicate that osmotic stress up-regulates expression of the AVT gene and increases the accumulation of AVT mRNA in the hypothalamus. This accumulation may, in part, be due to lengthening of the AVT mRNA poly(A) tail which is a general mechanism associated with stabilization of vertebrate mRNAs.

Characterization of porcine endometrial, myometrial, and mammary oxytocin binding sites during gestation and labor.

Changes in oxytocin (OT) receptor expression have been found to be an important determinant of the physiological effect of OT in several species. To date there are no published studies of OT binding sites during pregnancy in the pig. The purpose of the present study is to improve understanding of the role of OT in porcine parturition. The concentration and affinity of OT binding sites were determined for myometrium and endometrium from pregnant and postpartum gilts. Tissues were obtained after slaughter from 7 animals in each of four groups: 1) 90 days gestation, 2) 112 days gestation, 3) term after milk letdown (before delivery), and 4) within 1-3 h after farrowing. Mammary tissues were obtained for some animals in each group (n = 3-5/group). Before slaughter, blood was collected from each animal and assayed for estradiol-17 beta, progesterone, 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM), and OT. Binding of 3H-OT in the three tissues was concentration- and time-dependent. Sites of 3H-OT binding (fmol/mg protein +/- SEM) increased toward term for each tissue and remained elevated in the postpartum group. Endometrial and mammary tissues displayed the most acute increases in OT binding site concentrations while myometrial tissues displayed a more gradual increase in OT binding sites over the times studied. The binding sites displayed high affinity for 3H-OT and were characterized by linear Scatchard plots. Concentrations of estradiol-17 beta, PGFM, and OT (pg/ml +/- SEM) were positively correlated with 3H-OT binding site concentrations, whereas progestrone concentrations (ng/ml +/- SEM) were negatively correlated with binding site concentration, as determined by Pearson's Correlation Analysis. The data represent the first account of changes in the expression of OT binding sites on porcine tissues during gestation and labor.

Cardiovascular effects of angiotensin-II in chickens.

1. Mature WL cockerels with permanent cannulae in brachial artery and vein were restrained in an isolated sling. Blood pressure (BP) and heart rate (HR) were continuously recorded. When the chickens were habituated to the sling, injections began. In each experiment the cockerels were injected intravenously 6 times at 6 min intervals. 2. In the first experiment 6 injections of 0.5 nmol[Aspl, Val5]ANG-II/kg body weight were given. 3. In the second experiment oxytocin (OT) antagonist ([d(CH2)5-O-Me-Tyr2,Thr4,Tyr9,Orn8]VT) at a dose of 2 nmol/kg, was injected for the first 3 and 0.5 nmol ANG-II/kg for the last 3 injections. Such OT-antagonist pretreatment completely abolishes the vasodepressor (VDP) response to neurohypophysial peptides in chickens. 4. Injections of ANG-II resulted in a biphasic effect on BP, an initial brief fall followed by a prolonged rise. During the hypotensive phase, tachycardia developed which turned into bradycardia as the hypertensive phase appeared. No tachyphylaxis of the VDP effect of ANG-II was evident with repeated injections. 5. OT-antagonist pretreatment had no effect on the VDP response to ANG-II. 6. These results suggest that, unlike relaxation of chicken aortic ring in in vitro preparations, there is no tachyphylaxis of the VDP response to ANG-II, in vivo. Furthermore, the neurohypophysial peptides are not involved in the VDP effect of ANG-II because pretreatment with an OT-antagonist had no effect on it. The baroreflex buffers the effects of ANG-II on vascular tone by affecting HR. 7. As ANG-II is secreted during hypovolaemia, the biphasic haemodynamic response peptides may have a compensatory role following volume contraction.

Arginine vasotocin gene expression in neuroendocrine, reproductive and gastrointestinal tissues of the domestic fowl: detection by reverse transcriptase polymerase chain reaction.

Arginine vasotocin gene transcripts in various tissues of the domestic fowl were detected by reverse transcriptase polymerase chain reaction followed by Southern blot analysis using a 209 bp fragment from the 3'-region of a cDNA encoding chicken arginine vasotocin as the probe. Relatively strong signals were observed with hypothalamic, adenohypophysial and proventricular RNA as the starting material. Lesser signals were obtained from RNA isolated from shell gland, adrenal gland, post-ovulatory follicles and ovarian thecal cells. Arginine vasotocin gene transcripts were undetectable in the posterior pituitary gland, small intestine and large intestine. These results suggest that in addition to its well-known antidiuretic and oxytocic actions, arginine vasotocin may act as a local neuromodulator or mediator and have other important autocrine or paracrine actions in non-hypothalamic tissues.

Oxytocin antagonist blocks the vasodepressor but not the vasopressor effect of neurohypophysial peptides in chickens.

Cockerels with permanent cannulas in the brachial artery and vein were put into isolated slings. Arterial pressure and heart rate were continuously recorded. Following habituation, tests were initiated. In each cockerel 2 nmol/kg of the tested neurohypophysial peptide (NPs) or analogue was IV injected six times at 6-min intervals. Arginine vasotocin (AVT) caused an immediate vasodepressor (VDP) effect and tachycardia. These subsided within 20-30 s and were followed by a vasopressor (VP) response and bradycardia. On repeated injections of AVT, the VDP response declined and bradycardia intensified. Arginine vasopressin (AVP), oxytocin (OT), and mesotocin (MT) had short-lasting VDP effect in the following order of potency: OT = MT > AVT > AVP. Only AVT and, more effectively, AVP, caused a VP response. The VDP effect of MT and OT declined on repeated injections. When AVT was injected after three injections of MT, it had mostly an immediate VP effect. Although the V1 agonist is VP in chickens, at the dose used the V1 antagonist, [d(CH2)5,O-Me-Tyr2]AVP, had no effect on cardiovascular responses to AVT. Pretreatment with OT antagonist, [d(CH2)5-O-Me-Tyr2-Thr4.Tyr9.Orn8]VT, abolished the VDP effect of all NPs. Thus, MT had no effect on blood pressure, whereas AVP and, more effectively, AVT, had a marked immediate VP action. In chickens the VDP effect of NPs is probably mediated by an OT/MT-like receptor, wherein the peptide's ring structure, shared by AVT, OT, and MT, is important.(ABSTRACT TRUNCATED AT 250 WORDS)

An in situ hybridization and immunohistochemical study of vasotocin neurons in the hypothalamus of water-deprived chickens.

The distribution of immunoreactive vasotocin (IR-AVT) and AVT mRNA in the hypothalamus of White Leghorn cocks was determined by immunohistochemistry and in situ hybridization, respectively. In control birds that were provided with water ad lib, AVT mRNA was distributed in the periventricular and lateral regions of the hypothalamus in clusters of neurons that correspond structurally with the mammalian paraventricular (PVN) and supraoptic (SON) nuclei. Although the distribution of AVT, identified by immunohistochemistry of adjacent serial sections within the hypothalamus, was similar to the distribution of AVT mRNA, the possibility that some positive staining was due to mesotocin neurons was not excluded. Water deprivation for 2 and 4 days resulted in both an increase in levels of AVT mRNA per neuron and the number of AVT mRNA-containing cells. Additionally, water deprivation resulted in a decrease in the amount of IR-AVT per neuron. The results indicate that osmotic stimulation increases AVT gene expression not only in individual neurons but also by activating subpopulation of neurons that are not observed in normally hydrated birds.

At low dose, arginine vasotocin has vasopressor rather than vasodepressor effect in chickens.

It is widely accepted that both arginine vasopressin (AVP), and its avian analogue, arginine vasotocin (AVT) are vasopressor (VP) in mammals, but vasodepressor (VDP) in birds. The rat VP and chicken VDP assays were used to characterize neurohypophysial peptides and their analogues. However, these assays use pharmacological doses. Like AVP in mammals, AVT is secreted in birds during dehydration, when peripheral vasoconstriction rather than vasodilatation is needed. Mature White Leghorn cockerels with permanent cannulas in brachial artery and vein were put restrained in a sling. Mean arterial pressure (MAP); heart rate; and rectal, shank, skin, and comb temperatures were continuously recorded. The i.v. infusion regimen was: 1 hr of 0.05 ml/kg*min of saline followed by an hour of tested drug and another hour of saline. The drugs were: AVT, V1-agonist ([Phe2 Orn8]VT), and V2-agonist (DDAVP), in dosage of 50 pmol/kg*min each. AVT and to a lesser extent the V1-agonist increased MAP, caused bradycardia, and reduced thermal conductivity between core and skin or comb. The V2-agonist had no significant effect. Thus, at a low dosage AVT has a VP rather than a VDP effect. Bolus i.v. injection of 2 nmol AVT/kg caused an immediate and large drop in MAP accompanied by tachycardia which subsided within 20-30 sec. This was followed by a small rise in MAP and significant bradycardia. Bolus injection of 2 nmol/kg of either agonist had no hypotensive effect and only the V1-agonist caused the second-phase bradycardia.(ABSTRACT TRUNCATED AT 250 WORDS)

Evidence of a role for the turkey posterior pituitary in prolactin release.

The aims of this study were: (1) to examine whether the posterior pituitary contains prolactin releasing factor (PRF) activity, (2) to determine to what extent known neurohypophyseal peptides contribute to this activity, and (3) to compare posterior pituitary PRF activities of hens in different reproductive stages. Anterior pituitary cells derived from juvenile female turkeys were incubated with posterior pituitary extracts or test substances for 3 hr. Posterior pituitary extracts (0.1-0.8 equivalent) contained a potent substance(s) which stimulated PRL release in a concentration-dependent manner (2.4 +/- 0.08 to 6.5 +/- 0.23 micrograms/500 k cells). Arginine vasotocin (AVT) and vasoactive intestinal peptide (VIP) antisera (1:500) completely abolished the PRL-releasing activities of their respective peptides but partially reduced (P less than 0.05) the PRF activity of the posterior pituitary (AVT, 19.9%; VIP, 55.1%). Mesotocin antiserum did not alter (P greater than 0.05) PRL release induced by posterior pituitary extract. Posterior pituitary extract (0.01-0.5 equivalent) from hens in each of the various stages of the reproductive cycle induced a concentration dependent PRL release. The 0.5 posterior pituitary equivalent dose from reproductively quiescent (nonphotostimulated), laying, photorefractory, and incubating hens increased PRL release 2.4-, 2.9-, 3.8-, and 11.1-fold, respectively. The turkey posterior pituitary contains a potent PRF activity, partially accounted for by VIP and AVT, at the assayed concentrations, which varies with the reproductive cycle.

Embryonic differentiation of sexual dimorphism in vasotocin and mesotocin levels in chickens.

Chicken embryos of both sexes were injected on the tenth day of incubation with either estradiol benzoate (EB), aromatase inhibitor [1,4,6-androstatrien-3, 17-dione (ATD)], antiestrogen [tamoxifen (TAM)], antiandrogen [flutamide (FLU)], or the oil vehicle as control (C). At adulthood, at the age of 26 weeks, 10 chickens of each sex were killed and the amounts of immunoreactive arginine vasotocin (AVT) and mesotocin (MT) in the anterior hypothalamus (AHA), posterior hypothalamus (PHA), neurohypophysis (NHP), and pineal gland (PNL) were determined. Control hens had significantly more AVT in PNL and less MT in AHA and NHP than the corresponding roosters. This sexual dimorphism was affected by the embryonic treatments; TAM increased AVT in AHA of cockerels but not of hens. In both sexes, TAM and FLU increased AVT content in NYP. In males, but not in females, ATD also increased AVT content in the NHP. TAM and FLU administration to the female embryo reduced PNL AVT to the amount present in normal males. None of the treatments effected AHA MT in hens, while in cockerels TAM increased it. In females, TAM and FLU significantly increased NHP MT to the level of C males. In roosters, ATD, TAM, and FLU increased NHP MT further. In hens, but not roosters, FLU reduced MT in PNL. These results indicate that embryonic differentiation of the MT and AVT systems is affected by gonadal steroids in chickens.

Alterations in uterine contractility during the oviposition cycle in domestic hens.

1. The potencies of several neurohypophysial hormones were examined at different times during the oviposition cycle in an in vitro fowl oxytocic assay. 2. Uterine tissues were removed 2 h before (-2h-OP), immediately after (0h-OP) and 5 h after (+5h-OP) spontaneous oviposition. In addition, uterine tissue was removed immediately after oviposition was induced, by administering prostaglandin E2 2 h before an expected oviposition (Induced-OP). 3. The rank order of oxytocic potencies for the peptides was arginine vasotocin = vasopressin greater than oxytocin greater than mesotocin. The sensitivity of the uterus to the hormones was 0h-OP = Induced-OP greater than -2h-OP = +5-OP. 4. These results suggest that uterine sensitivity to neurohypophysial hormones changes during the oviposition cycle in domestic fowls.

Flight effects on plasma levels of neurohypophysial hormones in homing pigeons.

A significant increase in the circulating levels of arginine vasotocin (AVT), without change in plasma osmolality, was observed in homing pigeons immediately after a flight of 48 km, lasting 60-80 min. This increase in plasma AVT is perceived as part of an overall homeostatic mechanism of mobilizing lipid as fuel for the flight muscles, of water conservation and temperature regulation in the body during the homing flight within the specific distance and duration of flight and under normal weather conditions. However, there was no change in plasma levels of mesotocin (MT) which suggests that the birds were under no significant stress during the flight.

Effect of intrahypothalamic injections of norepinephrine and serotonin on plasma arginine vasotocin and mesotocin in the White Leghorn cockerel.

1. Saline (10 microliters), norepinephrine (NE) and Serotonin (5-HT), 500 nmol each, were injected into the anterior third ventricle (A3V; n = 7) or the posterior third ventricle (P3V; n = 11) of ananesthetised, unrestrained White Leghorn cockerels. Plasma arginine vasotocin (AVT) and mesotocin (MT) were measured 20, 60 and 120 min after injection. 2. Injection of NE into both the A3V and P3V had no significant effect on either plasma AVT or plasma MT at any of the sampling times. 3. Administration of 5-HT into the A3V significantly increased plasma MT about two-fold 20 min following injection. At 120 min time, plasma MT returned to normal. 4. In P3V birds, 5-HT had no effect on plasma MT in the first 20 min, but a significant increase in plasma MT occurred 60 to 120 min after injection. The magnitude of the response was lower than in the A3V cockerels. 5. Plasma AVT was not affected by 5-HT administration into the A3V at any of the sampling times, but 5-HT administration into the P3V caused significant rises in plasma AVT at 120 min. 6. Serotonergic, but not noradrenergic, induction of neurohypophysial peptide secretion was demonstrated.

Effects of sex and gonadal steroids on arginine vasotocin and mesotocin in the pineal gland and neurohypophysis of White Leghorn fowls.

1. Pineal and neurohypophysial arginine vasotocin (AVT) and mesotocin (MT) were measured in White Leghorn hens, cockerels and castrated males treated with either testosterone propionate (TP) or oestradiol benzoate (EB) (n = 10/group). The lighting regimen was 14 h light: 10 h dark, supplied by natural diffused sunlight and incandescent bulbs. 2. Both AVT and MT were detected in the pineal gland of all the chickens. 3. There was no significant effect of either sex or treatments on pineal MT. 4. Females had about 4 times more pineal AVT than males, regardless of their treatment. There was no effect of the treatments on pineal AVT in the males. 5. No sexual difference in neurohypophysial AVT was detected, but the neurohypophysis of the castrated males treated with EB contained less AVT than the neurohypophysis of the intact males. 6. Intact males had about twice as much MT in the neurohypophysis as females. Castrated males treated with either TP or EB had similar concentrations of neurohypophysial MT, which were lower than that of the intact males, but higher than that of the females.

Arginine vasotocin and mesotocin in the anterior hypothalamus, neurohypophysis, proventriculus and plasma of White Leghorn cockerels, during dehydration.

1. The effect of 96 hrs of water deprivation on plasma electrolytes, osmolarity, arginine vasotocin (AVT), mesotocin (MT), and on AVT and MT content in the neurohypophysis, anterior hypothalamic area (AHA) and proventriculus, was studied at 24 hrs intervals, in adult White Leghorn cockerels. 2. Plasma AVT increased three fold during the first 24 hrs but there was no further change during the next 48 hrs. In the last 24 hrs, plasma AVT decreased in about 25%. Plasma MT did not change during the entire period of dehydration. 3. Plasma sodium and osmolarity gradually increased during that time. 4. Neurohypophysial AVT content was depleted by 95% during the period of dehydration while MT content did not change. 5. In the AHA there was no change in AVT levels during dehydration while the levels of MT increased while in the proventriculus there was no change in either AVT or MT levels. 6. For the data collected during the entire experimental period, no correlation was found between plasma osmolarity and plasma AVT, but there was a highly significant negative correlation between plasma osmolarity and neurohypophysial AVT content. 7. It may be suggested that the depletion in AVT content in the neurohypophysis during progressive water deprivation resulted in an insufficient level of AVT in circulation to enable the cockerels to counter the dehydration. This may explain the death of those cockerels which were dehydrated for a further 24 hr period.

Arginine vasotocin and mesotocin levels in theca and granulosa layers of the ovary during the oviposition cycle in hens (Gallus domesticus).

Neurohypophysial hormones in mammals are synthesized by the ovary and may influence reproductive function. Although arginine vasotocin (AVT) can induce premature oviposition in chickens, the gonadal content of AVT or mesotocin (MT) has not been examined in this species. Ovarian follicles were removed 2 hr before oviposition (-2 hr), immediately after oviposition (0 hr), and 5 hr after oviposition (+5 hr). The theca and granulosa layers of the three largest preovulatory and postovulatory follicles were isolated, extracted, and assayed for immunoreactive AVT and MT. AVT content in the theca and granulosa layers was lowest at -2 hr and increased to maximum levels at +5 hr. Pre- and postovulatory follices contained similar amounts of AVT. Maximum AVT levels in the theca and granulosa layers were 2.5 +/- 0.6 and 1.3 +/- 0.2 ng/tissue, respectively. In contrast, theca content of MT was maximal at 0 hr and the highest peptide levels were observed in the largest preovulatory follicle (0.24 +/- 0.02 ng/tissue). The results demonstrate that, compared with circulating levels of the hormones, the ovary of the chicken contains high concentrations of AVT and MT. The ovarian content of AVT and MT varies in a different manner in relation to the oviposition cycle.