Release Of Oxytocin In Response Research Articles

Estimation of udder emptying based on milk constituents of strip samples after milking

Milk ejection disorders were induced by oxytocin receptor blockade. We tested the hypothesis that the degree of udder emptying at incomplete milk ejection can be estimated based on the concentration of various milk constituents in different milk fraction samples. To induce different levels of spontaneous udder emptying (SUE) 10 Holstein dairy cows were milked either with or without i.v. injection of the oxytocin receptor blocking agent atosiban (ATO). In ATOearly, 12 µg/kg BW ATO was injected immediately before and in ATOlate directly after a 1 min manual udder preparation. The normal milking routine served as Control treatment. In all 3 treatments the udder was completely emptied by the i.v. injection of 10 IU oxytocin (OT) at the end of spontaneous milk flow. During all experimental milkings 4 milk samples were taken in all treatments: at the start of udder preparation (foremilk; FM), immediately after cessation of spontaneous milk flow and cluster detachment by hand stripping (strip milk; SM), from spontaneous removed milk in the bucket 1 (milk before OT; MBOT) and from the milk obtained after OT injection in the bucket 2 (milk after OT; MAOT). Fat, protein, lactose and electrolytes (Na, Cl and K) were measured in each milk sample. In addition, electrical conductivity (EC) was determined in parallel to continuous milk flow recording. The treatments induced individual degrees of SUE; therefore, the final evaluations of data were based on SUE classes instead of treatments. The most pronounced differences of milk constituents at different degrees of SUE were found for the milk fat content. The fat content of SM and MBOT remained almost unchanged up to 60% SUE, but was considerably higher if >80% of the milk was spontaneously removed. The concentrations of Na and Cl were highest and of K lowest if less than 20% on milk was received in the different samples. The EC was higher in SM and MBOT if <20% of milk was received. In conclusion, the blockade of the OT effect influences primarily the fat content, which confirmed an OT-induced fat secretion during milking. Similar effects are likely found in situations of disturbed milk ejections, caused by a lacking or reduced release of OT in response to different degrees of tactile udder stimulation. Our results show that the measurement of fat content and the EC in strip milk samples collected after cluster detachment can be used to estimate the completeness of udder emptying.

Measuring oxytocin release in response to gavage: Computational modelling and assay validation.

In the present experiments, we tested the conclusion from previous electrophysiological experiments that gavage of sweet food and systemically applied insulin both stimulate oxytocin secretion. To do so, we measured oxytocin secretion from urethane-anaesthetised male rats, and demonstrated a significant increase in secretion in response to gavage of sweetened condensed milk but not isocaloric cream, and a significant increase in response to intravenous injection of insulin. We compared the measurements made in response to sweetened condensed milk with the predictions from a computational model, which we used to predict plasma concentrations of oxytocin from the published electrophysiological responses of oxytocin cells. The prediction from the computational model was very closely aligned to the levels of oxytocin measured in rats in response to gavage.

Endogenous oxytocin levels in extracted saliva elevates during breastfeeding correlated with lower postpartum anxiety in primiparous mothers

BackgroundBreastfeeding in the early postpartum period is expected to have mental benefits for mothers; however, the underlying psychobiological mechanisms remain unclear. Previously, we hypothesized that the release of oxytocin in response to the suckling stimuli during breastfeeding would mediate a calming effect on primiparous mothers, and we examined salivary oxytocin measurements in primiparous mothers at postpartum day 4 using saliva samples without extraction, which was erroneous. Thus, further confirmation of this hypothesis with a precise methodology was needed.MethodsWe collected saliva samples at three time points (baseline, feeding, and post-feeding) to measure oxytocin in 24 primiparous mothers on postpartum day 2 (PD2) and 4 (PD4) across the breastfeeding cycle. Salivary oxytocin levels using both extracted and unextracted methods were measured and compared to determine the qualitative differences. State and trait anxiety and clinical demographics were evaluated to determine their association with oxytocin changes.ResultsBreastfeeding elevated salivary oxytocin levels; however, it was not detected to a significant increase in the extraction method at PD4. We found a weak but significant positive correlation between changes in extracted and unextracted oxytocin levels during breastfeeding (feeding minus baseline); there were no other significant positive correlations. Therefore, we used the extracted measurement index for subsequent analysis. We showed that the greater the increase in oxytocin during breastfeeding, the lower the state anxiety, but not trait anxiety. Mothers who exclusively breastfed at the 1-month follow-up tended to be associated with slightly higher oxytocin change at PD2 than those who did not.ConclusionsBreastfeeding in early postpartum days could be accompanied by the frequent release of oxytocin and lower state anxiety, potentially contributing to exclusive breastfeeding.

Feeding and Manual Brushing Influence the Release of Oxytocin, ACTH and Cortisol Differently During Milking in Dairy Cows.

AimThis study aimed to examine the effects of feeding or abdominal brushing on the release of the hormones oxytocin, ACTH and cortisol during milking in dairy cows.MethodsTwelve cows in early lactation were used (2 × 2 factorial experimental design), testing the effects of two types of sensory stimulation during milking over a 3 day period; feeding concentrate or manual abdominal brushing (1 stroke/s). Blood samples for hormone analyses were collected at time at −15, −1, 0 (onset of cluster), every min for 8 min, at 10, 12, 14, 16, 30, and 60 min. Hormone levels were assayed and AUC was calculated.ResultsMilking was associated with an immediate and significant rise of oxytocin. When milking was combined with feeding, significantly higher levels of oxytocin were observed at 2 and 4 mins (p < 0.05). No effect of brushing on oxytocin levels was observed. Milking alone was associated with a significant rise of ACTH levels. Feeding in connection with milking reduced the immediate rise of ACTH levels (p < 0.05) and AUC (p < 0.02), whereas no effects of brushing were found. Milking caused a progressive rise of cortisol levels. Concomitant feeding did not influence cortisol levels, whereas brushing significantly decreased cortisol levels at 1, 5 and 14 mins after onset of milking (p < 0.05).ConclusionFeeding increases oxytocin release in response to milking and decreases ACTH levels. Abdominal brushing did not influence these variables, but decreased cortisol levels. These data demonstrate that activation of afferent vagal nerve fibres and of cutaneous sensory nerves originating from the abdominal skin in front of the udder influence milking related hormone release differently.

Vasopressin but Not Oxytocin Responds to Birth Stress in Infants.

ContextBirth triggers a large fetal neuroendocrine response, which is more pronounced in infants born vaginally than in those born by elective cesarean section (ECS). The two related peptides arginine vasopressin (AVP) and oxytocin (OT) play an essential role in peripheral and central stress adaptation and have a shared receptor mediating their function. Elevated cord blood levels of AVP and its surrogate marker copeptin, the C-terminal part of AVP prohormone, have been found after vaginal delivery (VD) as compared to ECS, while release of OT in response to birth is controversial. Moreover, AVP, copeptin and OT have not yet been measured simultaneously at birth.ObjectiveTo test the hypothesis that AVP but not OT levels are increased in infants arterial umbilical cord blood in response to birth stress and to characterize AVP secretion in direct comparison with plasma copeptin.MethodsIn a prospective single-center cross-sectional study, we recruited healthy women with a singleton pregnancy and more than 36 completed weeks of gestation delivering via VD or ECS (cesarean without prior uterine contractions or rupture of membranes). Arterial umbilical cord blood samples were collected directly after birth, centrifuged immediately and plasma samples were frozen. Concentrations of AVP and OT were determined by radioimmunoassay and that of copeptin by ultrasensitive immunofluorescence assay.ResultsA total of 53 arterial umbilical cord blood samples were collected, n = 29 from VD and n = 24 from ECS. Ten venous blood samples from pregnant women without stress were collected as controls. AVP and copeptin concentrations were significantly higher in the VD group than in the ECS group (both p < 0.001), median (range) AVP 4.78 (2.38–8.66) vs. 2.38 (1.79–3.88) (pmol/L), copeptin 1692 (72.1–4094) vs. 5.78 (3.14–17.97), respectively, (pmol/L). In contrast, there was no difference in OT concentrations (pmol/L) between VD and ECS, 6.00 (2.71–7.69) vs. 6.14 (4.26–9.93), respectively. AVP and copeptin concentrations were closely related (Rs = 0.700, p < 0.001) while OT did not show any correlation to either AVP or copeptin. In linear regression models, vaginal delivery and biochemical stress indicators, base deficit and pH, were independent predictors for both AVP and copeptin. OT was not linked to base deficit or pH.ConclusionVaginal birth causes a profound secretion of AVP and copeptin in infants. Whereas AVP indicates acute stress events, copeptin provides information on cumulative stress events over a longer period. In contrast, fetal OT is unaffected by birth stress. Thus, AVP signaling but not OT mediates birth stress response in infants. This unique hormonal activation in early life may impact neurobehavioral development in whole life.

92 Intra-nasal oxytocin treatment does not attenuate the hypothalamo-pituitary-adrenal axis in beef heifers subjected to isolation stress or restraint and isolation stress

Abstract Changes in the physiological, psychological, and behavioral manifestations of stress are observed in association with increases in circulating oxytocin. Providing oxytocin intra-nasally attenuates stressor-induced hypothalamo-pituitary-adrenal (HPA) axis activation in humans and rodents. The present study was conducted to investigate effects of intra-nasal oxytocin supplementation on stressor-induced activation of the HPA axis in beef cattle. We hypothesized that oxytocin would attenuate activation of the HPA axis, ultimately decreasing plasma cortisol. Bos taurus heifers (n = 28) were blocked by bodyweight and randomly allocated to one of four treatment groups, in a 2 × 2 factorial arrangement: 1) Saline, isolated, standing, and unrestrained (S-IS, 0.015mL/kg BW 0.9% isotonic saline, n = 7); 2) Saline, isolated, and restrained (S-RIS; 0.015 ml/kg BW 0.9% isotonic saline; n = 7); 3) Oxytocin, IS (OXT-IS, 0.3 IU/kg BW oxytocin; n = 7); and 4) Oxytocin and isolated and restrained (OXT-RIS, 0.3 IU/kg BW oxytocin; n = 7). Oxytocin and saline were administered intra-nasally. Appropriate stressors were applied 30 minutes following intra-nasal treatment. Blood samples were collected directly following imposing the stressor, and every 10 minutes thereafter, for 2 h. Cortisol concentrations increased over time in animals exposed to restraint and isolation stress (P &lt; 0.01) and decreased over time in animals exposed to isolation stress (P &lt; 0.01). In the present study, oxytocin did not affect measured indicators of HPA axis activation. Greater plasma oxytocin concentrations were observed in restrained animals administered saline, compared with their standing counterparts administered oxytocin (P &lt; 0.01), indicating some endogenous release of oxytocin in response to restraint stress. Overall, restraint stress increased cortisol and oxytocin in Bos taurus heifers compared with heifers subjected only to isolation. Finding a more intermediate stress model may allow for more precise detection of effects of oxytocin on the stress response.

Neuromodulation of maternal circuits by oxytocin.

Motherhood in mammals involves tremendous changes throughout the body and central nervous system, which support attention and nurturing of infants. Maternal care consists of complex behaviors, such as nursing and protection of the offspring, requiring new mothers to become highly sensitive to infant needs. Long-lasting neural plasticity in various regions of the cerebral cortex may enable the perception and recognition of infant cues, important for appropriate caregiving responses. Recent findings have demonstrated that the neuropeptide oxytocin is involved in a number of physiological processes, including parturition and lactation and dynamically shaping neuronal responses to infant stimuli as well. Here, we review experience-dependent changes within the cortex occurring throughout motherhood, focusing on plasticity of the somatosensory and auditory cortex. We outline the role of oxytocin in gating cortical plasticity and discuss potential mechanisms regulating oxytocin release in response to different sensory stimuli.

Technical note: Reduced pulsation chamber vacuum at normal pulsation rate and ratio provides adequate prestimulation to induce oxytocin release and milk ejection while simultaneous milk flow is prevented

In the present study we investigated the milking characteristics and the oxytocin release in dairy cows milked after either manual prestimulation or a premilking period with pulsating liners at normal pulsation rate (60) and ratio (60:40) while the pulsation chamber vacuum (PCV) was reduced to 20 kPa to prevent the opening of the liners. During the milking trial with 8 cows the PCV reduction was started either before attachment (PCV-1) or immediately after attachment (PCV-2) of the teat cups. Milk yields, total milking times, average milk flows, peak flow rates, the duration of milk flow plateaus, and the duration of milk flow declines did not differ among the 3 treatments. Only the time to reach peak milk flow was prolonged when the vacuum reduction was started after teat cup attachment (PCV-2). In this treatment, milk flow >200 g/min already occurred during the premilking period, resulting in bimodal milk flow curves. In 5 of the 8 cows, plasma oxytocin (OT) concentrations were measured from -2 min before the start of milking until 3 min of milking to compare the OT release in response to manual prestimulation and during PCV-1. In both treatments, OT increased similarly and remained elevated until the end of measurements. Consequently, the areas under the curve of OT concentrations did not differ between treatments. In conclusion, milking performance is similar if milking is performed after manual prestimulation or after normal pulsation at reduced PCV. To prevent milk flow during the prestimulation period, it is of crucial importance to start the reduction of the PCV before cluster attachment.

Forced swimming-induced oxytocin release into blood and brain: Effects of adrenalectomy and corticosterone treatment

The oxytocin (OXT) system is functionally linked to the HPA axis in a reciprocal and complex manner. Certain stressors are known to cause the simultaneous release of OXT and adrenocorticotrophic hormone (ACTH) followed by corticosterone (CORT). Furthermore, brain OXT attenuates ACTH and CORT responses. Although there are some indications of CORT influencing OXT neurotransmission, specific effects of CORT on neurohypophyseal or intra-hypothalamic release of OXT have not been studied in detail.In the present set of experiments, adult male rats were adrenalectomized (ADX) or sham-operated and fitted with a jugular vein catheter and/or microdialysis probe targeting the hypothalamic paraventricular nucleus (PVN). Blood samples and dialysates were collected before and after forced swimming (FS) and analyzed for CORT, ACTH and AVP concentrations (in plasma) and OXT concentrations (in plasma and dialysates). Experimental treatments included acute infusion of CORT (70 or 175μg/kg i.v.) 5min prior to FS, or subcutaneous placement of 40% CORT pellets resulting in stable CORT levels in the normal basal range.Although ADX did not alter basal OXT concentrations either in plasma or in microdialysates from the PVN, it did cause an exaggerated peripheral secretion of OXT and a blunted intra-PVN release of OXT in response to FS. CORT pellets did not influence either of these ADX-induced effects, while acute infusion of 175μg/kg CORT rescued the stress-induced rise in OXT release within the PVN and modestly increased peripheral OXT secretion.In conclusion, these results indicate that CORT regulates both peripheral and intracerebral OXT release, but in an independent manner. Whereas the peripheral secretion of OXT occurs simultaneously to HPA axis activation in response to FS and is modestly influenced by CORT, HPA axis activation and circulating CORT strongly contribute to the stress-induced stimulation of OXT release within the PVN.

PLASMA OXYTOCIN CONCENTRATION AND DEPRESSIVE SYMPTOMS: A REVIEW OF CURRENT EVIDENCE AND DIRECTIONS FOR FUTURE RESEARCH.

There is substantial recent interest in the role of oxytocin in social and affiliative behaviors-animal models of depression have suggested a link between oxytocin and mood. We reviewed literature to date for evidence of a potential relationship between peripheral oxytocin concentration and depressive symptoms in humans. Pubmed(®) and PsychINFO(®) were searched for biomedical and social sciences literature from 1960 to May 19, 2015 for empirical articles in English involving human subjects focused on the relationship between peripheral oxytocin concentration and depressive symptoms, excluding articles on the oxytocin receptor gene, or involving exogenous (i.e. intranasal) administration of oxytocin. Eight studies meeting criteria were identified and formally reviewed. Studies of pregnant women suggested an inverse relationship between oxytocin level and depressive symptom severity. Findings in nonpregnant women were broadly consistent with the role of oxytocin release in response to stress supported by animal studies. The relationship between oxytocin and depression in men appeared to be in the opposite direction, possibly reflecting the influence of gonadal hormones on oxytocinergic functioning found in other mammalian species. Overall, small sample sizes, heterogeneity in study designs, and other methodological limitations may account for inconsistent findings. Future research utilizing reliable oxytocin measurement protocols including measurements across time, larger sample sizes, and sample homogeneity with respect to multiple possible confounders (age, gender, race and ethnicity, ovarian status among women, and psychosocial context) are needed to elucidate the role of oxytocin in the pathogenesis of depression, and could guide the design of novel pharmacologic agents.

A double-blind randomized controlled trial of oxytocin nasal spray in Prader Willi syndrome.

Individuals with Prader-Willi syndrome (PWS) have a significant reduction in the number of oxytocin-producing neurons (42%) in the hypothalamic paraventricular nucleus. A number of animal studies and observations of humans show that lesions in this region can produce PWS-like symptoms. Given the evidence for potential oxytocin deficiency, we tested the effects of a course of intranasal oxytocin on PWS symptoms. Thirty individuals with PWS aged 12-30 years participated in an 18-week randomized double-blind placebo-controlled crossover trial. Participants received 8 weeks of oxytocin and 8 weeks of placebo with a minimum 2-week washout period. The first 11 participants received the following oxytocin doses: 24 IU (twice daily) B.I.D for participants 16 years and over and 18 IU B.I.D for participants 13-15 years. The dose was increased for the remaining 18 participants to 40 IU B.I.D for participants 16 years and over and 32 IU B.I.D for 13-15 years. Measures used to assess changes were standardized well-accepted measures, including the Developmental Behavior Checklist-Monitor, Parent, Teacher, and Adult; The Yale-Brown Obsessive Compulsive Scale; The Dykens Hyperphagia questionnaire; Reading The Mind in the Eyes Test; Epworth Sleepiness Scale and the Movie Stills. Oxytocin had little impact on any measure. The only significant difference found between the baseline, oxytocin, and placebo measures was an increase in temper outbursts (P = 0.023) with higher dose oxytocin. The lack of effect of oxytocin nasal spray may reflect the importance of endogenous release of oxytocin in response to exogenous oxytocin.

Kinetics of Oxytocin Response to Repeated Restraint Stress and/or Chronic Cold Exposure

Recently, several new atypical actions of circulating oxytocin are emerging, which may be of importance for the physiological effects of oxytocin released during stress. However, little information is available on oxytocin response to chronic stress stimuli. The aim of the present study is to deepen the knowledge on oxytocin secretion during chronic and repeated stress. The main hypothesis to be tested was that oxytocin release in response to single and to repeated or chronic stress is of different kinetics. Adult male Sprague-Dawley rats were exposed to 2 different stress stimuli or their combination. Restraint (immobilization) of different duration (10-120 min) and number of repetitions (1 or 7 times) as well as chronic exposure (28 days) to cold temperature were used. Concentrations of oxytocin in plasma and posterior pituitary were measured by a radioimmunoassay. Concentrations of oxytocin in plasma increased significantly in response to both single and repeated immobilization. Acute immobilization caused rapid increase already after 10 min of restraint, while the recovery occurred only after 24 h. Repeated restraint caused delayed onset of increased oxytocin release and a more rapid recovery to prestress levels after 3 h. In conclusion, the results of the present study show that though with a different kinetics, increased oxytocin release is preserved during repeated exposure to an intensive stressor, namely immobilization for 120 min. During repeated exposure to shorter stressors, an adaptation in oxytocin responses may occur. This should be taken into account with respect to cardiovascular and metabolic effects of stress-induced oxytocin.

State and Trait Affectionate Communication Buffer Adults' Stress Reactions

The present study tested the prediction that affectionate communication is positively associated with the release of oxytocin in response to stressors. One hundred participants completed questionnaire measures about their personal relationships prior to participation in a laboratory session that included a series of standard laboratory stressors. Both state and trait affectionate communication predicted increases in oxytocin during exposure to stressors, an effect that was not moderated by sex. The results demonstrate the stress-buffering effect of affectionate interaction.

Oxytocin release in magnocellular nuclei: neurochemical mediators and functional significance during gestation

When released from dendrites within the supraoptic (SON) and paraventricular (PVN) nuclei (intranuclear release) during suckling, oxytocin exerts autocrine and paracrine effects on oxytocin neurons that are necessary for the unique timing and episodic pattern of oxytocin release into the systemic circulation that is characteristic of lactation. Recent reports have shown that stimulation of central noradrenergic and histaminergic receptors are both necessary for intranuclear release of oxytocin in response to suckling. In addition, in vitro studies indicate that excitatory amino acids may also be critical for central oxytocin secretion, although in vivo experiments have not provided direct support for this hypothesis. In addition to a critical role in intranuclear oxytocin release during lactation, norepinephrine has also been shown to stimulate central oxytocin during gestation. Stimulation of central oxytocin receptors during gestation appears critical for normal systemic oxytocin secretion in responses to suckling during the subsequent period of lactation. Oxytocin receptor blockade during pregnancy alters normal timing of systemic oxytocin release during suckling and reduces milk delivery. Several adaptations occur in the central oxytocin system that are necessary for determining the unique response characteristic observed during parturition and gestation. Central oxytocin receptor stimulation during gestation has been implicated in pregnancy-related morphological changes in magnocellular oxytocin neurons, disinhibition of oxytocin neurons to GABA, and adaptations in membrane response characteristics of oxytocin neurons. In conclusion, intranuclear oxytocin release during gestation and lactation are critical for establishing, and then evoking the unique pattern of systemic oxytocin secretion in response to the suckling offspring necessary for adequate milk delivery. Furthermore, activation of central noradrenergic receptors appears to be critical for release of central oxytocin in both of these reproductive states.

A possible relationship between brain-derived adrenomedullin and oxytocin in the regulation of sodium balance

Exaggerated thirst and salt appetite occurs when endogenous, brain-derived adrenomedullin (AM) production is compromised. In addition, the arginine vasopressin (AVP) response to hypovolemia is compromised. We hypothesized that AM acts in the hypothalamus to control oxytocin (OT) release and that the inhibitory action of AM on salt appetite is mediated via its effects on OT release in the rat. When plasma tonicity was elevated with sodium, ribozyme-induced compromise of central AM production significantly blunted the release of OT into plasma. OT responses to elevation of plasma osmolality without concomitant change in plasma sodium levels were not altered by compromise of AM production. Thus, brain-derived AM controls OT release in response to altered plasma sodium levels. Furthermore, central AM-induced inhibition of NaCl intake can be reversed by pretreatment with an OT antagonist, and the increase in NaCl appetite seen following ribozyme compromise of central AM can be attenuated with central OT administration. These data support the hypothesis that endogenous, brain-derived AM is an essential participant in the hypothalamic response to hypernatremia via its actions on OT-expressing neurons. Together with our previous reports of the effects of AM on AVP secretion and ingestive behaviors, our results suggest that endogenous AM is a physiologically relevant regulator of the endocrine and behavioral mechanisms that maintain fluid and electrolyte homeostasis.

Evidence of Dysregulated Peripheral Oxytocin Release Among Depressed Women

Oxytocin is a hypothalamic neuropeptide that plays a key role in mammalian female reproductive function. Animal research indicates that central oxytocin facilitates adaptive social attachments and modulates stress and anxiety responses. Major depression is prevalent among postpubertal females, and is associated with perturbations in social attachments, dysregulation of the hypothalamic-pituitary-adrenal stress axis, and elevated levels of anxiety. Thus, depressed women may be at risk to display oxytocin dysregulation. The current study was developed to compare patterns of peripheral oxytocin release exhibited by depressed and nondepressed women. Currently depressed (N = 17) and never-depressed (N = 17) women participated in a laboratory protocol designed to stimulate, measure, and compare peripheral oxytocin release in response to two tasks: an affiliation-focused Guided Imagery task and a Speech Stress task. Intermittent blood samples were drawn over the course of two, 1-hour sessions including 20-minute baseline, 10-minute task, and 30-minute recovery periods. The 10-minute laboratory tasks did not induce identifiable, acute changes in peripheral oxytocin. However, as compared with nondepressed controls, depressed women displayed greater variability in pulsatile oxytocin release over the course of both 1-hour sessions, and greater oxytocin concentrations during the 1-hour affiliation-focused imagery session. Oxytocin concentrations obtained during the imagery session were also associated with greater symptoms of depression, anxiety, and interpersonal dysfunction. Depressed women are more likely than controls to display a dysregulated pattern of peripheral oxytocin release. Further research is warranted to elucidate the clinical significance of peripheral oxytocin release in both depressed and nondepressed women.

Angiotensin II-induced release of oxytocin: interaction with norepinephrine and role in lactation

These studies examined the receptors involved in angiotensin II (Ang II) stimulated secretion of systemic oxytocin (OT) and the role of this peptide in release of OT during suckling. Plasma OT concentrations were measured following intracerebroventricular (icv) injection of vehicle, Ang II, or Ang II following pretreatment with a selective AT1 (Losartan) or AT2 (PD 123319) receptor antagonist. Furthermore, we measured Ang II-induced OT release during central α-adrenergic receptor blockade (phentolamine). Finally, plasma OT concentrations before and during suckling were evaluated following central administration of Ang II receptor antagonists. The increase in systemic OT following central Ang II was abolished by AT1 receptor blockade and inhibited by the AT2 receptor antagonist. Furthermore, pretreatment with phentolamine significantly diminished systemic OT release in response to icv Ang II. Finally, central Ang II receptor blockade did not alter the increase in circulating OT during suckling. These data demonstrate that Ang II evoked OT release is mediated through activation of both AT1 and AT2 receptors and suggest that a component of Ang II-induced OT stimulation is due to norepinephrine release. Furthermore, central angiotensin systems do not have a direct role in stimulating OT release during suckling.

Endogenous opioid regulation of stress-induced oxytocin release within the hypothalamic paraventricular nucleus is reversed in late pregnancy: a microdialysis study

Oxytocin secretion into blood in response to swim stress is differentially regulated by endogenous opioids in virgin and pregnant rats. Here, the influence of endogenous opioids on oxytocin release within the hypothalamic paraventricular and supraoptic nuclei was investigated using microdialysis in virgin and pregnant (day 19–21) rats. Rats fitted with a U-shaped microdialysis probe 3 days before testing were injected with naloxone (5 mg/kg body weight, s.c.) or vehicle (sterile saline) and, 3 min later, were forced to swim (10 min at 19°C). Within the paraventricular nucleus, basal and stimulated oxytocin release did not significantly differ between vehicle-treated virgin and pregnant rats. After naloxone, local oxytocin release in response to swimming was lowered in virgin rats ( P<0.01), whereas it was further increased in pregnant rats ( P<0.01). Within the supraoptic nucleus, basal oxytocin release was significantly lower in pregnant compared to virgin rats ( P<0.01). Forced swimming induced a similar rise in intranuclear oxytocin release in both vehicle-treated virgin and pregnant rats, but peak levels were still higher in the virgin controls. In contrast to the paraventricular nucleus, naloxone did not alter swim-induced oxytocin release within the supraoptic nucleus either in virgin or pregnant rats. Vasopressin release in the paraventricular nucleus was also increased by forced swimming but there was no effect of pregnancy or naloxone on it. In summary, in pregnancy, basal and stress-induced oxytocin release within the paraventricular nucleus was not changed, whereas it was blunted within the supraoptic nucleus. Further, within the paraventricular nucleus the excitatory effect of endogenous opioids on local oxytocin release seen in virgins was switched into an inhibitory action in pregnancy. In contrast, endogenous opioids were evidently not involved in the regulation of swim-induced oxytocin release within the supraoptic nucleus either in virgin or pregnant rats. Thus, pregnancy-related neuroendocrine plasticity also includes site-specific functional alterations in opioid receptor-mediated actions in the hypothalamus.

Inhibition of oxytocin release and milk let-down in postpartum primiparous cows is not abolished by naloxone.

About 10% of primiparous cows have no milk ejection during the first milkings after delivery. Therefore, 17 Brown Swiss dairy cows in their first lactation were used to evaluate the extent of disturbed milk let-down and the corresponding oxytocin (OT) plasma values in the 1st 5 days after delivery. The first milking was 9-22 h after parturition and served for classification of the cows to groups with inhibited (INH), bimodal (BIMO), or normal (NOR) milk let-down. The OT plasma levels before the start of manual teat stimulation and machine milking were comparably high during the first milking especially in NOR and BIMO cows. Ten minutes before the second milking (M2), 300 mg of the opioid antagonist naloxone was injected to test whether the disturbance was affected by the action of endogenous opioids on the neurohypophysis. The milk yield was not influenced by the naloxone treatment, and the INH cows had milk ejection only after a vaginal stimulation. Afterwards, the cows were milked twice every day, until the milk let-down and the OT release were unaffected (equal to control milking). Then, at the next milking, the cows were injected with 300 mg morphine 10 min before milking. The central OT release in response to manual teat stimulation and machine milking was completely blocked in all cows, but a vaginal stimulation was able to abolish this block, at least partially, in 16 cows. Thus, morphine produced a milk let-down characteristic as in the INH cows during the first three milkings. For the following milking, the cows were pre-treated with 300 mg naloxone (-15 min) plus 300 mg morphine (-10 min) before milking. The OT release and the milk yields were unaffected when compared with the control milking. This experiment demonstrates that exogenous opioids can affect the central release of OT in a naloxone-reversible manner even very soon after parturition. However, endogenous opioids are probably not the main mediators of disturbed central OT release and alveolar milk ejection in post-partum primiparous cows.

Naloxone cannot abolish the lack of oxytocin release during unexperienced suckling of dairy cows

To evaluate the role of opioids for the regulation of oxytocin release in response to teat stimulation, 10 brown-Swiss dairy cows were randomized to two experiments during mid of lactation. In the first experiment, four cows without previous suckling experience were suckled by an alien calf between two normal milkings. Before and during milking or suckling, frequent blood samples were collected via a jugular cannula for determination of oxytocin and β-endorphin. In the second experiment, six cows were treated with naloxone or saline, 10 min before the start of the first or second suckling, respectively. The collected blood samples were assayed for oxytocin. In the first experiment, the plasma levels of β-endorphin were elevated during and after the unexperienced suckling in three cows, but not in the fourth cow, and the release of oxytocin during suckling was markedly reduced, suggesting no release of alveolar milk. In the second experiment, the release of oxytocin during suckling was again significantly reduced. Pretreatment with naloxone before suckling did not completely abolish the adverse effect of suckling and the oxytocin plasma level did not increase to levels comparable with control milking. In emotional stress situations, the release of oxytocin from the pituitary is inhibited with simultaneously elevated β-endorphin plasma levels. Although there is some evidence for a regulatory role of opioids for the release of oxytocin, other mediators are suggested to be more potent in regulating oxytocin under stress conditions.