Studies on the functional relationship between thyroid, adrenal and gonadal hormones

In order to clarify the functional relationship between thyroid, adrenal and gonadal hormones, hypothyroidism was induced by administration of thiuoracil in adult male and female rats, and the effects of hypothyroidism on the adrenal and the gonadal axes were investigated in the present study. 1. The functional relationship between thyroid and adrenal hormones: Adrenal weights and corticosterone were lowered, whereas the secretion of ACTH, corticotrophin-releasing hormone (CRH) and arginine vasopressin (AVP) increased in hypothyroid rats compared to euthyroid rats. These results indicate that hypothyroidism causes adrenal dysfunction directly and results in hypersecretion of CRH and AVP from the hypothalamus. 2. The functional relationship between thyroid and gonadal hormones: The pituitary response to LHRH was lowered, whereas the testicular response to hCG was not changed in hypothyroid rats. Hypothyroidism suppressed copulatory behavior in male rats. These results suggest that hypothyroidism probably causes dysfunction in gonadal axis at the hypothalamic-pituitary level in male rats. In adult female rats, hypothyroidism inhibited the follicular development accompanied estradiol secretion, whereas plasma concentrations of progesterone and prolactin (PRL) increased in hypothyroid female rats. Hypothyroidism significantly increased the pituitary content of vasoactive intestinal peptide (VIP) though it did not affect dopamine synthesis. These results suggest that hypothyroidism increases pituitary content of VIP and this increased level of VIP likely affects PRL secretion in a paracrine or autocrine manner. In female rats, inhibition of gonadal function in hypothyroid rats mediated by hyperprolactinemia in addition to hypersecretion of endogenous CRH.     

Regulation of adrenocorticotropin secretion from cultured canine anterior pituitary cells.

Pituitary cells, collected from five healthy dogs, were cultured and treated with various doses of ovine corticotropin-releasing hormone (CRH), arginine vasopressin (AVP), oxytocin (OT), or angiotensin II (AII) to determine which of these hypothalamic peptides affected adrenocorticotropin (ACTH) secretion. Of the 4 peptides, only CRH significantly increased ACTH secretion from cultured canine anterior pituitary cells. The lowest dose of CRH tested, 0.01 nM, significantly stimulated ACTH release. Co-addition of AVP, OT, or AII with CRH did not increase ACTH secretion beyond that caused by addition of CRH alone. Similarly, neither co-addition of AVP with OT, AVP with AII, or OT with AII significantly stimulated ACTH secretion. These results support a role for CRH in the physiologic regulation of ACTH secretion from the canine anterior pituitary, but do not support regulatory roles for AVP, OT, or AII.     

Distribution of the androgen receptor in the diencephalon and the pituitary gland in goats: Co-localisation with corticotrophin releasing hormone, arginine vasopressin and corticotrophs

Previously it has been shown that androgen suppresses transportation-induced increases in plasma adrenocorticotropic hormone (ACTH), possibly by suppressing the secretion of corticotrophin releasing hormone (CRH) or arginine vasopressin (AVP) from the hypothalamus, or secretion of ACTH from the pituitary gland. The aim of the present study was to examine androgen target sites in the caprine diencephalon and pituitary gland using immunohistochemical methods. The androgen receptor (AR) was expressed strongly in the bed nucleus of the stria terminalis, the medial preoptic area, the arcuate nucleus, the ventromedial hypothalamic nucleus and the suprachiasmatic nucleus in the diencephalon. Between 8% and 11% of CRH and AVP neurons in the paraventricular hypothalamic nucleus (PVN) expressed AR. In the pituitary gland, 7.1% of corticotrophs expressed AR. The results are consistent with the proposal that androgen acts directly and indirectly on CRH and/or AVP neurons in the PVN. The possibility of a direct action of androgen on the corticotrophs in the pituitary gland was also considered.     

The impact of maternal overnutrition and obesity on hypothalamic-pituitary-adrenal axis response of offspring to stress

We evaluated the effect of maternal obesity before and throughout gestation on offspring hypothalamic-pituitary-adrenal axis function. Multiparous Rambouillet by Columbia crossbred ewes were fed either 100% of National Research Council (NRC) recommendations (control, C) or 150% of NRC recommendations (obese, OB) from 60 d before mating until lambing. Ten lambs born to OB ewes (five males and five females), and eight lambs born to C ewes (three male and five female) were studied. From delivery to weaning lambs were maintained with their mothers, who were all fed 100% NRC recommendations. After weaning, all lambs were group housed and fed the same diet to meet NRC requirements. At 19 mo of age lambs were placed in individual pens and fed a pelletized diet to meet maintenance requirements. Jugular vein catheters were placed and 2 d later lambs received an intravenous (i.v.) adrenocorticotropic hormone (ACTH) challenge followed by an i.v. corticotropin-releasing hormone (CRH)/arginine vasopressin (AVP) challenge 1 d later. Thirty d later offspring were again catheterized and placed into metabolism crates for 2 d before receiving an isolation stress test. ACTH and cortisol responses to the isolation stress test and CRH/AVP challenge and cortisol responses to ACTH challenge were determined. Cortisol was quantified via radioimmunoassay and ACTH was quantified using an Immulite 1000; both were analyzed using repeated measures using the MIXED procedure of SAS. Offspring from OB ewes had elevated basal plasma ACTH and cortisol compared with C offspring before all three challenges (P < 0.05). Offspring from OB mothers tended (P = 0.06) to have a greater ACTH response after an i.v. CRH/AVP injection than offspring from C mothers (12,340 ± 1,430 vs 8,170 ± 1,570 area under the curve, respectively). Cortisol response to the CRH/AVP and ACTH challenges was not influenced by maternal nutrition (P = 0.46) and averaged 4.77 ± 0.2 μg/dL and 1.94 ± 0.01 μg/dL, respectively. The ACTH response following the isolation stress test was also similar (P = 0.82) for OB and C offspring (147 ± 20 pg/mL), and cortisol response during the isolation stress test was similar between C and OB offspring (P = 0.64, 5.25 ± 0.3 μg/dL). These findings suggest that maternal obesity before and during gestation does not affect stress responses by the offspring, but has an impact on hypothalamic-pituitary-adrenal sensitivity. The lack of differences in cortisol release under the influence of difference concentrations of ACTH during the CRH/AVP challenge could indicate adrenal dysfunction in OB offspring.     

促肾上腺皮质激素释放激素结合蛋白功能与调控研究进展

促肾上腺皮质激素释放激素结合蛋白(CRH-BP)是37 ku的分泌糖蛋白,其结构与促肾上腺皮质激素释放激素(CRH)受体不同,但与CRH和尿皮素(UCN)具有很高的亲和力.CRH-BP具有重要的生物调控作用,对CRH诱导的脑垂体促肾上腺皮质激素(adrenocorticotropic hormone,ACTH)的释放具有抑制作用.应激和代谢因素能改变体内CRH-BP的表达与调控,CRH-BP在生产分娩、炎性疾病、阿尔海默症(Alzheiner′s disease)和其他机能障碍等过程中也具有广泛的作用,这说明CRH-BP在大脑和脑垂体中可能还有其他潜在功能,研究CRH-BP在中枢和外周的作用及机理,有利于阐明其在病因学方面的潜在地位,有助于治疗CRH失调引起的机能紊乱.     

Decreased ACTH secretion during prolonged transportation stress is associated with reduced pituitary responsiveness to tropic hormone stimulation in cattle

The present study examined the effect of transportation stress on hypothalamic–pituitary–adrenal axis responsiveness to tropic hormone stimulation and on abundance of corticotropin releasing factor (CRF) receptor R1 (CRFR1) and arginine vasopressin (AVP) receptor V3 (V3) mRNAs in the anterior pituitary (AP) of cattle. Holstein steers were transported for 10 h or used as non-transported controls (NTC). Blood samples were collected at start of transportation and every 1–2 h thereafter. To test AP responsiveness to tropic hormones, animals were challenged (i.v.) with CRF (0.5 μg/kg), AVP (1 μg/kg) or CRF plus AVP immediately after end of transportation and blood samples collected every 30 min for 3 h. The AP of animals transported for 0, 4 or 10 h were harvested for mRNA analyses. Plasma ACTH in transported animals increased within 1 h and remained elevated for 6 and 8 h versus NTC and 0 h values, respectively. Plasma concentrations of cortisol increased in response to transportation and remained elevated throughout the transport period. Injection of CRF or AVP to NTC animals increased plasma ACTH, but ACTH secretion in response to CRF or AVP was dramatically reduced in transported animals. ACTH secretion following co-injection of CRF and AVP tended to be less in transported animals, but was almost 100% greater than when secretagogues were administered separately. Despite decreased AP responsiveness to CRF and AVP, AP CRFR1 and V3 mRNAs were increased after 10 h transportation. Results indicate decreased AP responsiveness to CRF and AVP may regulate duration of ACTH secretion in response to transportation stress in cattle.     

Hormonal interactions within the hypothalamus and pituitary with respect to stress and reproduction in sheep

Endocrine systems may be used as indicators of stress in two ways. The primary role of a hormone may be as part of the homeostatic response to a stimulus (e.g., adrenaline, corticosteroids). The amplitude of hormone response may correlate with the severity of the stimulus and any change indicate that the body is responding. Alternatively, a hormone may have a key role in normal body function (e.g., reproduction) and stress may deleteriously alter the hormone signal prevent normal function. This demonstrates that the stimulus was sufficiently severe that homeostatic mechanisms were unable to maintain normal function. Stress may effect reproduction by reducing both LH pulse amplitude and frequency. The LH surge may also be delayed. Several mechanisms may account for these effects both at the hypothalamus and pituitary. Corticosteroids have a broad, yet fundamental, role in homeostasis and have been used as primary indicators of stress for many years. Excess corticosteroid can be detrimental so the concentration is controlled via the hypothalamus-pituitary-adrenal (HPA) axis by multi-level feedback mechanisms. Under field and experimental conditions, after an initial large response prolonged stimulation leads to a gradually reducing plasma corticosteroid concentrations. This has been interpreted as a reduction in perceived stimulus severity or habituation to the stimulus and the animal deemed "less stressed" and its welfare "better." However, this reduction may be due to the intrinsic control mechanisms designed to prevent prolonged increases in corticosteroid concentrations. The stress signal at higher brain levels may still be present and the animal may still be experiencing the stimulus as aversive. Thus, the welfare interpretation of a corticosteroid concentration may differ during the time course of a stress response. A greater understanding of the mechanisms controlling corticosteroid secretion at each level of the HPA is required to determine what is the correct interpretation at any time point. To address these issues, we have used mathematical modelling to produce representations of possible control mechanisms at each level of the HPA. The starting point was to measure AVP and CRH concentrations in hypophysial portal blood and ACTH and cortisol concentrations in jugular blood in conscious sheep during 2h road transport (a cognitive stimulus). Modelling identified the signal inputs that were most likely to explain the secretion rate of each hormone. Modelling suggested that the reduction in AVP and CRH secretion observed during transport was most likely due to a reduction in stimulus input, with a significant contribution from cortisol negative feedback only on AVP secretion. At the pituitary level, ACTH secretion was stimulated more by AVP than by CRH (ratio 2.3:1) and there was also a stimulatory effect related to cortisol concentration at the time of sampling. However, the responses to both stimuli were curtailed by cortisol negative feedback and an inhibitory effect of prior CRH concentration. These are complex effects, but the modelling does suggest that while "stress" inputs may reduce over time hormone negative feedback is a major factor reducing hormone responses. When interpreting hormone data for animal welfare purposes, it is important not to interpret a reduction in hormone concentration due to intrinsic hormone control mechanisms as a reduction due to a decrease in the stress stimulus.     

Adrenocorticotropic hormone-induced secretion of cortisol in goats is inhibited by androgen

In a previous study, it was found that there are sex differences in goats with respect to the levels of cortisol secretion induced by transportation stress. We also found that treatment of castrated male goats with dihydrotestosterone (DHT) suppressed the increase in plasma cortisol concentration following transportation, but did not suppress the secretion of adrenocorticotropic hormone (ACTH). This suggests that androgen might block ACTH ‐ induced cortisol secretion. In order to examine this hypothesis, the effects of androgen on ACTH‐induced cortisol secretion in goats were investigated. First, castrated male goats were treated with testosterone (T), DHT or cholesterol (cho) for 21–25 days. Cho was used as a control for T and DHT treatment. Then, plasma cortisol concentrations were compared among the hormonal treatments after ACTH injection. Subsequently, the distribution of androgen receptors in the caprine adrenal gland was investigated. There were no differences in the basal cortisol concentrations among the hormonal treatments. However, plasma cortisol concentrations after ACTH injection were significantly lower in T ‐ and DHT ‐ treated goats than in cho ‐ treated goats. Androgen receptors were present in 60% of the cells in the zonae fasciculata and reticularis of the adrenal cortex, the regions that secrete glucocorticoids. These results suggest that androgen may act directly on the adrenal cortex to suppress cortisol secretion induced by ACTH.     

Restraint effects on stress-related hormones and blood natural killer cell cytotoxicity in pigs with a mutated ryanodine receptor

A mutation in the ryanodine receptor gene (RYR1) of the calcium release channel is responsible for increased stress susceptibility in pigs. In the present study, the relation of a mutation in RYR1 with the neuroendocrine (stress-related hormone) response and the immune defense represented by natural killer cell cytotoxicity (NKCC) during a 4-h restraint and recovery phase in 60 male pigs was investigated. Blood samples were collected from pigs previously divided into RYR1 genotypes (nn, Nn, NN), based on PCR amplification and restriction analyses. The blood samples collected during the restraint and recovery phases of the experiment were used to determine NKCC (⁵¹Cr-release assay), large granular lymphocyte number (hematologic method), and plasma concentrations of prolactin (PRL), GH, ACTH, and cortisol (COR) (by specific RIA). The greatest degree of NKCC response (P < 0.05) to restraint stress relative to controls was observed for the stress-susceptible homozygote group (nn). Measures of stress-related hormones were positively correlated with NKCC during the entire experimental period (P < 0.001 for all investigated hormones) in the nn group. Immunostimulatory effects in the early (0–60 min) phase of restraint were associated with increased hormone responses, especially PRL and GH. In the late (180–240 min) phase of stress and the recovery phase (480 min), a decrease in immune response was accompanied by an elevated COR response in all RYR1 genotypes. Moreover, divergent responses of both PRL (greatest in nn, P < 0.001) and GH (greatest in NN, P < 0.001) to the 4-h restraint were observed. Our results suggest that stress-susceptible RYR1-mutated homozygotes develop a greater level of immune defense, including cytotoxic activity of NK cells, and accompanied by more pronounced stress-induced changes in neuroendocrine response than stress-resistant heterozygous (Nn) and homozygous (NN) pigs.     

In vitro and in vivo temporal aspects of ACTH secretion: stimulatory actions of corticotropin-releasing hormone and vasopressin in cattle

The objective of the present study was to evaluate the temporal aspects associated with corticotropin-releasing hormone (CRH) and vasopressin (VP) stimulated bovine adrenocorticotropic hormone (ACTH) secretion in vitro and in vivo. For the in vitro studies, bovine anterior pituitary glands were enzymatically dispersed to establish primary cultures. On day 5 of culture, cells were challenged for 3 h with medium alone (Control) or various combinations and concentrations of bovine CRH (bCRH) and VP. Both CRH and VP each increased (P < 0.05) ACTH secretion. Maximal increases in ACTH secretion occurred in response to 0.1 microM CRH (5.5-fold) and 1 microM VP (3.7-fold), relative to Control cells. The in vivo portion of the study examined possible temporal differences in the activation of the pituitary-adrenal axis by CRH and VP. Jersey cows were randomly assigned to one of four groups (n = 8 cows/group): (i) Control (saline); (ii) bCRH (0.3 microg/kg BW); (iii) VP (1 microg/kg BW) and (iv) bCRH (0.3 microg/kg BW) + VP (1 microg/kg BW). Jugular blood samples were collected at 15-min intervals for 4 h pre- and for 6 h post-treatment; samples were also taken at 1, 5 and 10 min post-treatment. Plasma concentration of ACTH did not differ among treatment groups for the 4-h pre-treatment period. At 1 min post-treatment, bCRH + VP, VP and bCRH increased ACTH secretion by 22.4-, 9.6- and 2.2-fold, respectively, relative to Control (32.7 +/- 7.2 pg/ml). Maximal plasma concentration of ACTH occurred at 5, 10 and 15 min post-treatment for the VP (1017.7 +/- 219.9 pg/ml), bCRH + VP (1399.8 +/- 260.1 pg/ml) and bCRH (324.8 +/- 126.2 pg/ml) treatment groups respectively. Both the in vitro and in vivo data demonstrated that while VP acutely activates the bovine pituitary-adrenal axis, CRH-induced ACTH secretion is slower in onset but of longer duration. The present study also provides insight into the dynamics of ACTH and cortisol (CS) responsiveness to CRH and VP in cattle.     

Effects of corticotropin-releasing hormone,lysine vasopressin,oxytocin, and angiotensin II on adrenocorticotropin secretion from porcine anterior pituitary cells

The aim of this study was to determine the ability of corticotropin-releasing hormone (CRH), lysine vasopressin (LVP), oxytocin (OT), and angiotensin II (AII) to stimulate adrenocorticotropin (ACTH) secretion from porcine anterior pituitary (AP) cells in vitro and to evaluate the role of protein kinase C (PKC) in the interaction between CRH and LVP. In this study, porcine AP cells were enzymatically and mechanically dispersed, cultured (150,000 cells/well) for 4 d, and then challenged with doses of various neuropeptides for 3 hr. CRH (10⁻⁷−10⁻¹⁰ M) was the most potent of the peptides tested in stimulating ACTH release from porcine AP cells. In fact, none of the other peptides consistently affected ACTH concentrations relative to basal levels. However, LVP potentiated CRH action, even though by itself, it failed to stimulate ACTH production. Neither OT or AII potentiated CRH-stimulated ACTH release from porcine AP cells. To determine whether the interaction between CRH and LVP was regulated partially by the protein Kinase C (PKC) pathway, we challenged AP cells in a 30-min incubation with 10⁻⁷ M staurosporine (ST), a treatment predicted to decrease PKC activity. Then, cells were washed and challenged with 10⁻⁹ M LVP, 10⁻⁹ M CRH, and 10⁻⁹ M CRH + LVP. Treatment with ST decreased (P < 0.05) CRH + LVP-stimulated ACTH release. To further demonstrate an interaction between protein kinase A (PKA) and PKC transduction pathways in the observed synergism between CRH and LVP to enhance ACTH secretion, we also challenged AP cells with 10⁻⁷ M phorbol 12, 13-myristate acetate (PMA) and 5 μM forskolin (FOR) for 3 hr. This treatment was predicted to enhance PKA and PKC activities, respectively, and thereby enhance ACTH concentrations. Challenging cells with FOR + PMA enhanced (P < 0.001) ACTH release above basal concentrations, but more important, it increased (P < 0.001) ACTH concentration above that elicited by either drug given alone. Taken together, our in vitro studies support the conclusion that CRH is the principal regulator of ACTH secretion in the pig. In contrast to the results in most other species evaluated, vasopressin alone did not affect ACTH release. However, LVP can enhance the effectiveness of CRH in releasing ACTH, and this enhancement appears to rely, at least in part, on the activation of the PKC signal transduction pathway.     

Dopamine Antagonist Affects Luteal Function But Not Cyclicity During the Autumn Transition

The autumn transition is characterized by a number of anomalies affecting the luteal phase of the estrous cycle, such as declining progesterone secretion from the corpus luteum and increased rate of failure of luteolysis. Prolactin also decreases. We theorized that these events may be linked, possibly through dopamine. We administered domperidone, a specific dopamine D2 receptor antagonist from September 12 to anestrus or January 15, either daily (n = 8), or once per day for the first 7 days of each month (n = 7). Controls (n = 7) received no treatment. Mean progesterone and prolactin concentrations were compared with summer cycles. Time to entry into anestrus and incidence of spontaneously prolonged luteal activity was compared between groups during the autumn transition. Prolactin declined from June through October equally in all groups. There was no difference between groups in time to anestrus. Progesterone decline was prevented (daily treatment) or delayed for a prolonged period (weekly group) in autumn. The incidence of spontaneously prolonged corpus luteum activity was reduced to summer levels in both domperidone groups compared with the control. We concluded that autumn prolactin decline was not driven through dopaminergic D2 receptor inhibition of pituitary lactotrophs. Sustained progesterone synthesis through the autumn may be the result of action of the D2 receptor antagonist with dopamine receptors on the corpus luteum. Autumnal luteolytic function appears to have a dopamine-influenced component. There does not appear to be a causative relationship between autumnal progesterone and prolactin secretion.     

Adrenocorticotropic hormone and cortisol in calves after corticotropin-releasing hormone.

The aim for this study was to analyze responsiveness of the hypothalamo-pituitary-adrenocortical axis to exogenous bovine corticotropin-releasing hormone (bCRH) in calves. Two dose-response studies were carried out, using either bCRH alone (dose rates of 0, .01, .03, and .1 microg bCRH/kg live weight) or in combination with arginine-vasopressin (bCRH:AVP, 0:0, .1:.05, .5:.25, and 1:.5 microg kg live weight). The bCRH was administered i.v. to calves (n = 5 to 7 per dose) housed individually or in groups. Serial blood samples were obtained from before to 300 min after injection and analyzed for plasma ACTH and cortisol concentrations. The lowest bCRH dose that produced a response in all calves was .1 microg/kg. In the experiment using bCRH with AVP, increasing the bCRH dose from .1 to 1 microg/kg resulted in an increase in peak ACTH concentration (321 vs. 2,003 pg/mL) but did not significantly affect the peak cortisol concentration (37 vs. 40 ng/mL). The time to reach the peak cortisol concentration increased with the dose of bCRH with AVP (from 38 to 111 min). The ACTH and cortisol concentrations determined at any time between 20 and 90 min after bCRH injection were correlated to the integrated responses calculated as areas under the ACTH and the cortisol curves (r between .61 and .99, P<.05). In comparison with results from studies in humans, pigs, and sheep, our data showed that the pituitary of calves seems less sensitive to CRH than that of other mammals, despite a greater capacity to produce ACTH. Moreover, the calf's adrenals seem to have a lower capacity to produce cortisol than adrenals of other mammals. As in other species, it seems that AVP enhances the release of ACTH and cortisol. For CRH challenge to be used in calves, we suggest injecting at least .1 microg of bCRH/kg live weight either with or without AVP and taking several blood samples before injection and between 20 and 90 min after injection.     

Physiological and behavioural effects of an intracerebroventricular injection of corticotropin releasing hormone in goats

This study investigated the effects of an intracerebroventricular (ICV) injection of corticotropin releasing hormone (CRH) on physiological and behavioural responses in goats. In Experiment 1, saline (control) or saline plus 25 microg of ovine CRH was injected into the third ventricle of castrated male goats. CRH increased plasma cortisol (Cor) levels markedly within 15 min, but had little effect on plasma glucose (Glu). Compared with saline injected goats, CRH decreased the total duration of lying behaviour but increased its frequency, and suppressed rumination and self-grooming. In Experiment 2, the effects of an intravenous (IV) injection of human adrenocorticotropic hormone (ACTH) (1-24) (0.1mg) were examined and an IV injection of saline was used as control. ACTH increased plasma Cor levels markedly, but did not change any behaviour compared with controls. It was concluded that CRH mediated the response of the hypothalamus-pituitary-adrenal (HPA) axis and behaviour following stress in goats, although the CRH-induced behavioural changes were independent of the HPA axis and seemed to be the result of direct action within the central nervous system.     

Integration Between Different Hypothalamic Nuclei Involved in Stress and GnRH Secretion in the Ewe

This study investigated possible integrated links in the neuroanatomical pathways through which the activity of neurones in the paraventricular nucleus and arcuate nucleus may modulate suppression of gonadotrophin‐releasing hormone (GnRH) secretion during stressful situations. Double‐label immunofluorescence and laser scanning confocal microscopy were used to examine the hypothalamic sections from the follicular phase ewes. Noradrenergic terminals were in close contact with 65.7 ± 6.1% corticotrophin‐releasing hormone (CRH) and 84.6 ± 3.2% arginine vasopressin (AVP) cell bodies in the paraventricular nucleus but not with β‐endorphin cell bodies in the arcuate nucleus. Furthermore, γ‐amino butyric acid (GABA) terminals were close to 80.9 ± 3.5% CRH but no AVP cell bodies in the paraventricular nucleus, as well as 60.8 ± 4.1%β‐endorphin cell bodies in the arcuate nucleus. Although CRH, AVP and β‐endorphin cell terminals were identified in the medial pre‐optic area, no direct contacts with GnRH cell bodies were observed. Within the median eminence, abundant CRH but not AVP terminals were close to GnRH cell terminals in the external zone; whereas, β‐endorphin cells and terminals were in the internal zone. In conclusion, neuroanatomical evidence is provided for the ewe supporting the hypothesis that brainstem noradrenergic and hypothalamic GABA neurones are important in modulating the activity of CRH and AVP neurones in the paraventricular nucleus, as well as β‐endorphin neurones in the arcuate nucleus. These paraventricular and arcuate neurones may also involve interneurones to influence GnRH cell bodies in medial pre‐optic area, whereas the median eminence may provide a major site for direct modulation of GnRH release by CRH terminals.     

Effect of ovary lipid of skipjack tuna on rat serum components after stress application

Lipid extracted from the ovary of skipjack tuna by the method that we developed is rich in phospholipid-type docosahexaenoic acid. The ovary lipid of skipjack tuna (OLS) was studied for its anti-stress activity in male Wistar rats, focusing on stress-related blood components: recovery from stress was examined after application of water immersion restraint stress. As a result, serum corticosterone (CORT) secretion was inhibited and decreased rapidly after stress application in rats given OLS compared with control rats. As CORT acts as a glucocorticoid, non-esterified fatty acid (NEFA) is expected to increase by stress application. However, the concentration tended to be lower in rats given OLS than in control rats. With respect to OLS concentration, OLS increased serum dehydroepiandrosterone, secretion concentration-dependently. In addition, as with the recovery study, it tended to inhibit the increase in NEFA. These results indicate that OLS may have an anti-stress activity against acute stress.     

Adenohypophyseal function in bitches treated with medroxyprogesterone acetate

The aim of this study was to investigate the effects of treatment with medroxyprogesterone acetate (MPA) on canine adenohypophyseal function. Five Beagle bitches were treated with MPA (10mg/kg, every 4 weeks) and their adenohypophyseal function was assessed in a combined adenohypophyseal function test. Four hypophysiotropic hormones (CRH, GHRH, GnRH, and TRH) were administered before and 2, 5, 8, and 11 months after the start of MPA treatment, and blood samples for determination of the plasma concentrations of ACTH, cortisol, GH, IGF-1, LH, FSH, prolactin, alpha-MSH, and TSH were collected at -15, 0, 5, 10, 20, 30, and 45 min after suprapituitary stimulation. MPA successfully prevented the occurrence of estrus, ovulation, and a subsequent luteal phase. MPA treatment did not affect basal and GnRH-induced plasma LH concentrations. The basal plasma FSH concentration was significantly higher at 2 months after the start of MPA treatment than before or at 5, 8, and 11 months after the start of treatment. The maximal FSH increment and the AUC for FSH after suprapituitary stimulation were significantly higher before treatment than at 5, 8, and 11 months of MPA treatment. Differences in mean basal plasma GH concentrations before and during treatment were not significant, but MPA treatment resulted in significantly elevated basal plasma IGF-1 concentrations at 8 and 11 months. MPA treatment did not affect basal and stimulated plasma ACTH concentrations, with the exception of a decreased AUC for ACTH at 11 months. In contrast, the maximal cortisol increment and the AUC for cortisol after suprapituitary stimulation were significantly lower during MPA treatment than prior to treatment. MPA treatment did not affect basal plasma concentrations of prolactin, TSH, and alpha-MSH, with the exception of slightly increased basal plasma TSH concentrations at 8 months of treatment. MPA treatment did not affect TRH-induced plasma concentrations of prolactin and TSH. In conclusion, the effects of chronic MPA treatment on adenohypophyseal function included increased FSH secretion, unaffected LH secretion, activation of the mammary GH-induced IGF-I secretion, slightly activated TSH secretion, suppression of the hypothalamic-pituitary-adrenocortical axis, and unaffected secretion of prolactin and alpha-MSH.     

Involvement of neurokinin receptors in the control of pulsatile luteinizing hormone secretion in rats

It has recently been shown that neurokinin B, a tachykinin, is associated with GnRH pulse generation in sheep and goats. The aim of the present study was to clarify the role of tachykinin receptors in the control of LH secretion in rats. To this end, we evaluated the effect of CS-003, an antagonist for all three neurokinin receptors (NK1, NK2 and NK3 receptors), on pulsatile LH secretion in both sexes of rats with different routes of administration. Both oral and third ventricular administration of CS-003 suppressed LH secretion in both sexes of gonadectomized animals. Furthermore, intact male rats with oral administration of CS-003 showed decreased serum testosterone levels, which might be due to suppressed LH secretion. None of the three subtype-specific neurokinin receptor antagonists showed a significant effect on LH secretion in ovariectomized rats when each antagonist was singly administered. The present results suggest that neurokinins play a role in the control of pulsatile GnRH/LH secretion via multiple neurokinin receptors in both male and female rats.     

Administration of ACTH to restrained, pregnant sows alters their pigs' hypothalamic-pituitary-adrenal (HPA) axis

This study was designed to examine the physiology and behavior of pigs whose dams were snared and then injected with ACTH during gestation. Administration of ACTH to dams during pregnancy has been shown to replicate the effects of prenatal stress in other species. Control sows (n = 8) were given no treatment, whereas the treatment sows (ACTH, n = 8) were immobilized by snaring the snout and then administered an i.v. injection of ACTH (1 IU/kg BW) weekly from 6 to 12 wk of gestation. A pig was killed from each sow at 1, 30, and 60 d of age. The hypothalamus, pituitary gland, adrenal glands, and liver were immediately obtained to determine the amounts of corticotropin-releasing hormone (CRH), beta-endorphin, and mRNA for pro-opiomelanocorticotropin (POMC), the ACTH receptor (ACTH-R), and insulin-like growth factor I (IGF-I). Pituitary corticotrope and somatotrope cell numbers and adrenal cortex-to-medulla area ratios (CORT:MED) were also determined. Pigs' behaviors were recorded at 6 and 8 wk of age. At 75 d of age, a blood sample was taken and a biopsy puncture was created on one pig from each litter, then pigs were stressed by mixing. Blood was sampled every other day for 10 d to determine plasma cortisol concentrations and differential leukocyte counts. Biopsy damage was evaluated for healing. At 1 d of age, control pigs tended to weigh more (P = .09), have a lower expression of ACTH-R mRNA (P = .01) and IGF-I mRNA (P = .01), and a lower CORT:MED (P = .04) than ACTH pigs. At 30 d of age, control pigs had a greater concentration of beta-endorphin (P = .01) and tended to have a lower concentration of CRH (P = .09) and IGF-I mRNA (P = .10) than ACTH pigs. At 60 d of age, control pigs tended to have lighter pituitary glands (P = .08), a lower expression of POMC mRNA (P = .02), and a CORT:MED (P = .003) than ACTH pigs. At 8 wk of age, control pigs performed a higher frequency of belly nosing (P = .07) and oral vice behaviors (P = .01) than ACTH pigs. In response to mixing stress, control pigs had lesser concentrations of plasma cortisol (P = .03) and healed faster (P = .006) than ACTH pigs. Thus, exogenous ACTH and restraint during gestation alters the HPA axis of the sow's offspring, and during stressful situations later in life health, and therefore welfare, may be compromised.     

The role of neurokinin A and its receptor in the regulation of prolactin secretion by the anterior pituitary of cyclic pigs

In pigs, plasma prolactin concentration markedly changes during the oestrous cycle and the regulation of its secretion is very complex. The contribution of neurokinins in this process has not been sufficiently delineated. The aim of the study was to examine the effects of neurokinin A (NKA) on prolactin synthesis and secretion in cyclic gilts. The expression of NKA precursor (Ppta) and receptor (Tacr2) genes as well as NKA and TACR proteins content in the porcine pituitaries (days 2–3, 9–10, 12–13, 15–16 and 19–20 of the cycle) was determined. Furthermore, the in vitro influence of NKA on the expression of prolactin (Prl), dopamine receptor (D2r), TRH receptor (Trhr) genes and prolactin secretion by the porcine pituitary cells (days 9–10, 15–16 and 19–20 of the cycle) was assessed. The expression of Ppta and Tacr2 as well as NKA and TACR proteins in the pituitary tissue has been changing throughout the oestrous cycle. NKA affected in vitro the expression of studied genes and prolactin secretion depending on the stage of the cycle, dose of NKA and/or duration of the cell incubation. Altogether, the study indicates that NKA is engaged in the modulation of prolactin secretion in the pig during the oestrous cycle.