Effects of brief exposure of male pheromone on multiple-unit activity at close proximity to kisspeptin neurons in the goat arcuate nucleus

Exposure of females to the male pheromone induces pulsatile release of gonadotropin-releasing hormone (GnRH) in goats. Recently, kisspeptin neurons in the arcuate nucleus (ARC) have been suggested to represent the proximate source of the GnRH pulse generator. In this study, we examined the effects of the pheromone on multiple-unit activity (MUA) in female goats fitted with recording electrodes aimed at the ARC kisspeptin neurons. In all eight goats, periodic bursts in MUA (MUA volleys), which were considered to be electrophysiological manifestations of the GnRH pulse generator, were observed. The mean intervolley interval (T) during the control period was calculated in each goat that was then exposed to the male pheromone for 1 sec at timings of 1/4 T, 1/2 T or 3/4 T after one regularly occurring MUA volley. An instantaneous rise in MUA was observed immediately after the exposure regardless of timing. Exposure at a timing of 3/4 T resulted in an MUA volley within 60 sec following the instantaneous rise in all goats. In contrast, an MUA volley was induced in only 2 goats by exposure at 1/2 T, while exposure at 1/4 T failed to induce an MUA volley in any goats. These results suggest that transmission of the pheromone signal to the ARC, represented by an instantaneous rise, activates the GnRH pulse generator. Moreover, the timing-dependent pheromone action in inducing an MUA volley indicates that the GnRH pulse generator has a refractory period for the pheromone signal after the burst.     

Effects of intravenous administration of neurokinin receptor subtype-selective agonists on gonadotropin-releasing hormone pulse generator activity and luteinizing hormone secretion in goats

Recent evidence suggests that neurokinin B (NKB), a member of the neurokinin (tachykinin) peptide family, plays a pivotal role in gonadotropin-releasing hormone (GnRH) pulse generation. Three types of neurokinin receptors (NKRs), NK1R, NK2R and NK3R, are found in the brain. Although NKB preferentially binds to NK3R, other NKRs are possibly also involved in NKB action. The present study examined the effects of intravenous administration of the NKR subtype-selective agonists GR73632 (NK1R), GR64349 (NK2R), and senktide (NK3R) on GnRH pulse generator activity and luteinizing hormone (LH) secretion. Multiple-unit activity (MUA) was monitored in ovariectomized goats (n = 5) implanted with recording electrodes. Characteristic increases in MUA (MUA volleys) were considered GnRH pulse generator activity. Although three NKR agonists dose-dependently induced an MUA volley and an accompanying increase in LH secretion, the efficacy in inducing the volley markedly differed. As little as 10 nmol of senktide induced an MUA volley in all goats, whereas a dose of 1000 nmol was only effective for the NK1R and NK2R agonists in two and four goats, respectively. When the treatment failed to evoke an MUA volley, no apparent change was observed in the MUA or LH secretion. Similar effects of the NK2R and NK3R agonists were observed in the presence of estradiol. The results demonstrated that NK3R plays a predominant role in GnRH pulse generation and suggested that the contributions of NK1R and NK2R to this mechanism may be few, if any, in goats.     

Local administration of Neurokinin B in the arcuate nucleus accelerates the neural activity of the GnRH pulse generator in goats

Kisspeptin neurons in the arcuate nucleus (ARC), which co-express neurokinin B (NKB) and dynorphin A, are termed KNDy neurons. These neurons are candidates for the intrinsic gonadotropin-releasing hormone (GnRH) pulse generator. The central and peripheral administration of NKB or its receptor (NK3R) agonist evokes GnRH pulse generator activity and the subsequent pulsatile GnRH/luteinizing hormone (LH) secretion. However, the mechanism responsible for neural activation of the GnRH pulse generator in goats is unclear. We conducted electrophysiological and histochemical experiments to test the hypothesis that KNDy neurons receive NKB and that the signal is transmitted bilaterally to a population of KNDy neurons. Bilateral electrodes aimed at a cluster of KNDy neurons were inserted into the ovariectomized goat ARC. We observed the GnRH pulse generator activity, represented by characteristic increases in multiple-unit activity (MUA volleys). The unilateral administration of NKB or vehicle in the close vicinity of KNDy neurons under simultaneous MUA recording from both sides revealed that only NKB evoked MUA volley(s) immediately after administration. The timing of the MUA volley(s) evoked on the ipsilateral side was synchronized to that on the contralateral side. The double-labeled ISH for KISS1 and TACR3, which encode kisspeptin and NK3R, respectively, revealed that most KNDy neurons co-expressed TACR3. Therefore, NKB could directly stimulate KNDy neurons, following which the stimulatory signal is immediately transmitted to the entire population of KNDy neurons via connection with their fibers. This mechanism helps synchronize burst activity among KNDy neurons, thereby generating neural signals that govern pulsatile GnRH secretion.     

Electrophysiological and Morphological Evidence for Synchronized GnRH Pulse Generator Activity Among Kisspeptin/Neurokinin B/Dynorphin A (KNDy) Neurons in Goats

Neurons in the arcuate nucleus (ARC) that concomitantly express kisspeptin, neurokinin B (NKB) and dynorphin A are termed KNDy neurons and are likely candidates for the intrinsic gonadotropin-releasing hormone (GnRH) pulse generator. Our hypothesis is that KNDy neurons are functionally and anatomically interconnected to generate discrete neural signals that govern pulsatile GnRH secretion. Our goal was to address this hypothesis using electrophysiological and anatomical experiments in goats. Bilateral electrodes targeting KNDy neurons were implanted into ovariectomized goats, and GnRH pulse generator activity, represented by characteristic increases in multiple-unit activity (MUA volleys), was measured. Spontaneous and pheromone- or senktide (an NKB receptor agonist)-induced MUA volleys were simultaneously recorded from both sides of the ARC. An anterograde tracer, biotinylated dextran amine (BDA), was also injected unilaterally into the ARC of castrated male goats, and the distribution of fibers containing both BDA and NKB was examined using dual-labeling histochemistry. The results showed that MUA volleys, regardless of origin (spontaneous or experimentally induced), occur simultaneously between the right and left sides of the ARC. Tract tracing indicated that axons projecting from NKB neurons in the ARC were directly apposed to other NKB neuronal cells located bilaterally in the ARC. These results demonstrate that GnRH pulse generator activity occurs synchronously between both sides of the ARC in goats and that KNDy neurons are bilaterally interconnected in the ARC via NKB-containing fibers. Taken together, the results suggest that KNDy neurons form a neuronal circuit to synchronize burst activity among KNDy neurons and thereby generate discrete neural signals that govern pulsatile GnRH secretion.     

Androgen induces production of male effect pheromone in female goats

Previously we showed that the primer pheromone responsible for the "male effect" was produced in specific skin regions of castrated male goats by androgen treatments. In the present study, we examined whether androgen can also induce production of the male effect pheromone in female goats. Capsules containing dihydrotestosterone (DHT) or testosterone (T) were subcutaneously implanted into six ovariectomized (OVX) goats for 28 days. Small skin samples were collected from the head and rump regions, and the pheromone activity of their ether extracts was examined using a bioassay that monitors the electrophysiological manifestation of the hypothalamic gonadotropin-releasing hormone pulse generator as multiple-unit activity. Behaviors of OVX goats towards ovary-intact estrous goats were also examined before and at the end of DHT or T treatment. Before androgen treatment, neither the head nor rump skin samples in OVX goats showed pheromone activity. DHT treatment induced pheromone activity in the head skin sample of six OVX goats and in the rump skin sample of two OVX goats. Similar results were obtained by T treatment. In addition, OVX goats treated with T showed masculine-type sexual behaviors such as courtship and mounting behaviors towards the estrous goats. These results demonstrate that androgen is capable of inducing primer pheromone activity in the female and suggest that the synthesis pathway of the male effect pheromone exists in both sexes in the goat.     

Effects of Gonadotrophin Releasing Hormone Administration on the Pituitary-Ovarian Axis in Anoestrous vs Ovariectomized Bitches

The aim of this study was to determine the effects of gonadotrophin releasing hormone (GnRH) administration on the plasma concentrations of reproductive hormones in intact and ovariectomized (OVX) bitches. Therefore, blood samples were collected at multiple times before and after the administration of 10 microg/kg GnRH (Fertagyl)) for the determination of the plasma concentrations of luteinizing hormone (LH), oestradiol, progesterone and testosterone in six anoestrus and in six OVX bitches. The mean plasma LH concentrations before and 60 min after GnRH administration were significantly lower in the anoestrous bitches than in the OVX bitches. In both groups GnRH administration resulted in a significant increase in the plasma LH concentration. The highest plasma LH concentrations were found at 10 min after GnRH administration and these values did not differ significantly between the two groups. Only in the anoestrous bitches a significant increase in plasma oestradiol concentrations was found after GnRH administration and these values were significantly higher than those in the OVX bitches. The plasma concentrations of progesterone and testosterone were low (close to or below the limit of quantitation) both before and after GnRH administration and the differences between anoestrous and OVX bitches were not significant. It can be concluded that (i) basal plasma LH concentration is significantly higher in OVX bitches than in anoestrous bitches, (ii) plasma LH concentration increases after GnRH administration in both anoestrous and OVX bitches, (iii) GnRH administration causes a significant rise in plasma oestradiol concentration only if ovarian tissue is present and (iv) measurement of plasma progesterone and testosterone concentrations before and after GnRH administration does not aid in distinguishing between anoestrous and OVX bitches. The results of this study may provide a basis for the diagnosis of remnant ovarian tissue and verification of neuter status in the bitch.     

Effects of fasting and refeeding on plasma concentrations of leptin, ghrelin, insulin, growth hormone and metabolites in swine

The effects of nutritional status, such as fasting and refeeding, on leptin and ghrelin secretion in swine were examined. The swine (n = 4) were fasted for 54 h and plasma hormone levels were measured before, during and after fasting. Plasma leptin and insulin concentrations began to decrease 12 and 6 h into the fasting period, respectively (P < 0.05), and maintained a low level for the remaining period of fasting. Plasma leptin and insulin returned to the pre‐fasting value 6 and 12 h after refeeding, respectively. Plasma ghrelin concentrations showed a nocturnal periodicity during the fasting period; it increased nocturnally at 36 and 42 h into the fasting period (P < 0.05). Plasma growth hormone levels did not show any remarkable changes during the fasting. Plasma glucose levels showed a modest fall during fasting and significantly decreased (P < 0.05) at 24 h into the fasting period, returning to pre‐fasting levels after refeeding. Plasma nonesterified fatty acid levels increased (P < 0.05) at 12 h into the fasting period and returned to the pre‐fasting level 6 h after refeeding. These results indicate that plasma leptin, insulin and ghrelin play an important role in maintaining energy homeostasis in swine. The plasma ghrelin did not continuously increase, but showed nocturnal periodicity during fasting. This may suggest that ghrelin is also involved in physiological processes other than energy homeostasis.     

Urinary excretion of purine derivatives and plasma allantoin level in sheep and goats during fasting

The relationship between blood plasma level and urinary excretion of allantoin (AN) was examined in sheep and goats during fasting to investigate the possible use of purine derivatives (PD) in urine and/or plasma for estimating the microbial protein production in the rumen, and the further digestion in the lower guts of ruminants. Urinary AN excretion decreased markedly during fasting (0.13 mmol/kgW^0.75 per day), although urinary levels of other PD, hypoxanthine + xanthine and uric acid did not differ irrespective of the feeding condition, that is, feeding, fasting and refeeding in both species. The AN concentration in blood plasma also decreased drastically in the starvation period, and was suddenly increased on refeeding in sheep and goats, and these phenomena were very similar to those of urinary AN excretion. Therefore, there was a high positive correlation between plasma AN level and urinary AN excretion, and the coefficient of correlation was statistically significant (P < 0.01). These results clearly indicate that changes in urinary AN reflect change in plasma AN, which is induced by the catabolism of purine base in the body.     

Hormonal and metabolic adaptation to fasting: effects on the hypothalamic-pituitary-ovarian axis and reproductive performance of rabbit does

To assess the impact of acute caloric shortage on reproduction, rabbit does were either fed ad libitum (control, AL), or fasted for 24 (STF) or 48 h (LTF) before induction of ovulation with GnRH injection. Blood samples were collected during the last 3 h of fasting, and the following 4 h after GnRH injection, when feed was provided again, to measure plasma concentrations of LH, estradiol-17beta, leptin, insulin, T3, corticosterone, glucose, and NEFA. Before re-feeding, plasma leptin, insulin, and T3 concentrations were lower (P < or = 0.01) in both fasted groups than in controls, but then gradually increased following realimentation to match those of controls. During fasting, corticosterone levels were higher (P < or = 0.01) in LTF than in STF and AL does, but decreased to control values soon after realimentation. During fasting, plasma glucose concentrations did not differ among groups, but upon re-feeding they markedly increased (P < or= 0.01) both in STF and LTF does. NEFA levels were also more elevated (P < or = 0.01) in fasted rabbits than in controls, and rapidly decreased (P < or = 0.01) after re-feeding. Following GnRH injection, LH peak was lower (P < or = 0.01) in LTF than in AL and STF does. Estradiol-17beta showed higher pulse frequency and amplitude in AL than in STF and LTF does. Compared to controls, receptivity rate of STF and LTF artificially inseminated does declined respectively by -20.5% (P < or = 0.05) and -22.7%, and fertility rate by -23.9% (P < or = 0.05) and 21.4%, but no difference was found in ovulation rate. In summary, nutritional status of does, as modified by fasting, greatly influenced fertility, metabolic and reproductive hormones.     

Effect of exogenous corticosterone in genetically fat and lean chickens

1. The effects of daily injections of corticosterone (1 or 5 mg/bird) on growth, fat deposition, liver lipid and plasma concentrations of uric acid, glucose, insulin and growth hormone were studied using genetically selected lines of fat (FL) and lean (LL) chickens. 2. Both doses of corticosterone depressed body weight gain and increased the liver lipid and the abdominal fat to the same extent in both lines. 3. In both lines, corticosterone caused a dose-dependent increase in the plasma concentrations of uric acid, glucose and insulin in the fasted and refed states. 4. In untreated birds, plasma concentrations of growth hormone (GH) were slightly higher in FL than in LL chickens and slightly decreased during refeeding. The response was not modified by injection of 1 mg corticosterone. Injections of 5 mg decreased plasma GH in both lines in the fasting state and in LL chickens during refeeding. In contrast, the same dose increased GH in FL chickens during refeeding. This contradiction remains unexplained. 5. The results suggest that corticosterone sensitivity is not involved in difference of fattening between FL and LL chickens.     

Changes in metabolic and reproductive characteristics associated with lactation and glucose infusion in the postpartum ewe

This study examined mechanisms whereby the metabolic environment interacts with basic reproductive function. Ewes lambing during the breeding season were fed to maintain (MAINT, n = 10) or gain (GAIN, n = 11) body weight during the last 4 mo of gestation. From d 7 to 22 postpartum, ewes were infused iv with saline (n = 10) or glucose at a rate calculated to increase normal glucose entry rate by 75% (n = 11). Blood samples were collected daily to determine plasma concentrations of nutritive metabolites and insulin and at frequent intervals on d 14 and 21 to determine serum gonadotropin concentrations. Hypothalami and pituitaries were collected on d 22 to determine hormone content and receptor concentrations. Plasma concentrations of nutritive metabolites and insulin indicated that MAINT ewes mobilized more (P less than .01) body fat and protein reserves during gestation and early lactation than ewes in the GAIN group. Glucose infusion elevated plasma concentrations of glucose (P less than .05) and insulin (P less than .07) and reduced (P less than .05) fat and protein mobilization, even though it depressed feed intake (P less than .001), compared with saline infusion. Hypothalamic gonadotropin-releasing hormone (GnRH), pituitary luteinizing hormone (LH) and follicle-stimulating hormone (FSH) content and pituitary GnRH receptor concentration were similar between treatments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Origin of plasma insulin-like growth factor-I (IGF-I) during estrus in goats

The objective of this study was to clarify the origin of the increase in plasma insulin-like growth factor-I (IGF-I) during estrus in goats. Focusing on the uterus, the effect of estradiol-17 beta (E2) on the secretion of IGF-I was examined using ovariectomized and hysterectomized animals. A single 5 microg/kg BW of E2 was injected intramuscularly into ovariectomized and hysterectomized goats for 3 consecutive days, and plasma IGF-I concentrations in the two groups were compared. The concentrations of IGF-I rose after the treatments in both groups. The concentrations were significantly higher from 3 to 8 days after the treatment than before the treatment in ovariectomized goats (P<0.05), and from 1 to 3 days after the treatment than before in hysterectomized goats (P<0.05). Thus higher concentrations of plasma IGF-I tended to last longer in ovariectomized than hysterectomized goats. The area under the IGF-I response curve for the 8-day period after the first injection of E2 tended to be greater in ovariectomized than in hysterectomized goats. The results show that E2 increases plasma IGF-I concentrations in goats, and suggest that E2-stimulated IGF-I in plasma may originate mainly from the uterus.     

Effect of short-term fasting on plasma concentrations of leptin and other hormones and metabolites in dairy cattle

We determined the effects of short-term fasting and refeeding on temporal changes in plasma concentrations of leptin, insulin, insulin-like growth factor- 1 (IGF-1), growth hormone (GH), glucose, and nonesterified fatty acids (NEFA), in early lactating cows, non-lactating pregnant cows, and postpubertal heifers. In experiment 1, Holstein cows in early lactation were either fed ad libitum (Control, n=5) or feed deprived for 48 h (Fasted, n=6). Plasma leptin, insulin, and glucose concentrations rapidly declined (P<0.05) within 6h, and IGF-1 by 12h, but all these variables sharply returned to control levels (P>0.10) within 2h of refeeding. Plasma NEFA and GH concentrations were elevated (P<0.05) by 4 and 36 h of fasting and returned to control levels (P>0.10) by 8 and 24h after refeeding, respectively. In experiment 2, four ruminally cannulated pregnant non-lactating Holstein cows were used in a cross-over design and were fasted for 48 h (Fasted) or fasted with partial evacuation of rumen contents (Fasted-Evac). The plasma variables measured did not differ (P>0.10) between Fasted and Fasted-Evac cows. Plasma leptin, insulin, and IGF-1 concentrations were reduced by 10, 6, and 24h of fasting, respectively, in Fasted-Evac cows; and these variables were reduced by 24h in Fasted cows (P<0.05). Plasma glucose levels were reduced (P<0.05) by 48 h of fasting in both groups of fasted animals. Plasma NEFA and GH levels were increased (P<0.05) by 12 and 48 h of fasting, respectively. In experiment 3, postpubertal Holstein heifers were either fed ad libitum (Control, n=4) or feed deprived for 72 h (Fasted, n=5). Concentrations of leptin, insulin, IGF-1, and glucose in plasma were reduced (P<0.05) by 24, 10, 24, and 48 h of fasting, respectively. Plasma NEFA concentrations increased (P<0.05) by 4h, of fasting while GH levels were not significantly (P>0.10) affected by fasting. Collectively, our data provide evidence that plasma leptin concentrations are reduced with short-term fasting and rebound on refeeding in dairy cattle with the response dependent on the physiological state of the animals. Compared to the rapid induction of hypoleptinemia with fasting of early lactation cows, the fasting-induced hypoleptinemia was delayed in non-lactating cows and postpubertal heifers.     

Dynorphin-Kappa Opioid Receptor Signaling Partly Mediates Estrogen Negative Feedback Effect on LH Pulses in Female Rats

Accumulating evidence suggests that the arcuate nucleus (ARC) kisspeptin/neurokinin B (NKB)/dynorphin (KNDy) neurons play a role in estrogen negative feedback action on pulsatile gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) release. The present study aimed to determine if dynorphin (Dyn) is involved in estrogen negative feedback on pulsatile GnRH/LH release. The effect of the injection of nor-binaltorphimine (nor-BNI), a kappa-opioid receptor (KOR) antagonist, into the third cerebroventricle (3V) on LH pulses was determined in ovariectomized (OVX) adult female rats with/without replacement of negative feedback levels of estradiol (low E₂). The mean LH concentrations and baseline levels of LH secretion in nor-BNI-injected, low E₂-treated rats were significantly higher compared with vehicle-treated controls. On the other hand, the nor-BNI treatment failed to affect any LH pulse parameters in OVX rats without low E₂ treatment. These results suggest that Dyn is involved in the estrogen negative feedback regulation of pulsatile GnRH/LH release. The low E₂ treatment had no significant effect on the numbers of ARC Pdyn (Dyn gene)-,Kiss1- and Tac2 (NKB gene)-expressing cells. The treatment also did not affect mRNA levels of Pdyn and Oprk1 (KOR gene) in the ARC-median eminence region, but significantly increased the ARC kisspeptin immunoreactivity. These findings suggest that the negative feedback level of estrogen suppresses kisspeptin release from the ARC KNDy neurons through an unknown mechanism without affecting the Dyn and KOR expressions in the ARC. Taken together, the present result suggests that Dyn-KOR signaling is a part of estrogen negative feedback action on GnRH/LH pulses by reducing the kisspeptin release in female rats.     

Endocrine responses in mares undergoing abrupt changes in nutritional management

Leptin, a protein hormone secreted by adipocytes, plays an important role in energy homeostasis and regulation of body composition. We previously observed that acute feed restriction resulted in a rapid decline in concentrations of leptin in obese pony mares. This acute response prompted us to characterize the temporal changes in concentrations of leptin, GH, and insulin in obese pony mares during the transition between fed and feed-restricted conditions. Nine obese pony mares of mixed breed, previously maintained on fescue pasture, were randomly allotted to 2 groups. Treatments consisted of a 48-h feed restriction, a 48-h refeeding, and a 24-h feed restriction (RFR; n = 4), or 48 h of alfalfa hay ad libitum, a 48-h feed restriction, and a 24-h refeeding (FRF; n = 5). Blood samples were taken every 15 min during restriction and feeding transitions (0600 to 1400 on d 2 and 4), and every 30 min thereafter until 0830 of the following days (d 3 and 5). In the FRF treatment, plasma concentrations of leptin declined precipitously 6 h after the removal of feed (sample by treatment interaction; P < 0.01), and remained low and unresponsive to refeeding. Similarly, in the RFR group, plasma concentrations of leptin were initially low, and did not respond to feeding during the second (refeeding) sampling period. After feed restriction in each of the 2 treatment sequences, plasma insulin decreased and GH mean concentration, pulse frequency, pulse amplitude, and area under the curve increased (P < 0.05). Refeeding reversed these effects on insulin and GH. These data provide evidence that peripheral concentrations of insulin and GH are dynamically responsive to feed removal (decrease in insulin; increase in GH) and replacement (increase in insulin; decrease in GH), whereas leptin decreases in response to feed restriction but is slow to recover from a transient nutritional insult.     

Neuroendocrine control of reproductive function in ruminants

Gonadotropin‐releasing hormone (GnRH) is a key molecule in the control of reproduction in mammals. It is generally thought that the secretion of GnRH into the pituitary portal vessels is governed by two distinct neural mechanisms: the pulsatile and surge mode centers. The former is called the GnRH pulse generator, and this neural substrate plays a role as the master regulator of the reproductive function. An electrophysiological technique for monitoring the neural activity of the GnRH pulse generator has been established in the Shiba goat. The central actions of several neuropeptides have been assessed using this system. Results suggest that several neuropeptides including neuropeptide Y, cholecystokinin‐octapeptide and melanocortins are involved in the regulation of the GnRH pulse generator activity in the goat. Each input of those neuropeptides likely represents a unique mechanism conveying specific information about changes in the internal and external environments such as olfactory signals, nutrition, stress, and steroidal milieu, to the GnRH pulse generator. Further elucidations of actions of neurotransmitters on the GnRH pulse generator may serve for better understanding of the neuroendocrine control of reproduction in the ruminant.     

Different responses of protein synthesis to refeeding in various muscles of fasted chicks

1. The change in the rate of protein synthesis of different muscles, concentrations of plasma insulin, plasma insulin-like growth factor-I (IGF-I) and other plasma components were investigated after refeeding in fasted chicks. 5.2 g of the complete diet was refed. This was the maximum that could be force-fed with water. 2. The fractional synthesis rates (FSR) of breast (M. pectoralis major) and leg (M. gastrocnemius) muscles were measured after injection of L-[2, 6-(3)H]phenylalanine. Plasma insulin and IGF-I concentration were determined by radioimmunoassay. 3. In the breast muscle, FSR was significantly reduced by 2-d fasting. The FSR had recovered completely after 1 h of refeeding and was maintained until 6 h. The change in FSR after refeeding was associated with the change in ribosomal efficiency (K(RNA); absolute synthesis rate per unit RNA), while no change in ribosomal capacity (C(S); RNA: protein ratio) was observed. 4. In the leg muscle, FSR was decreased by 2-d fasting and increased gradually toward 6 h after refeeding but did not reach the level of the fed control. In contrast to the breast muscle, no significant changes in Cs and K(RNA) in the leg muscle were observed. 5. Plasma glucose concentration increased significantly at 1 h after refeeding but returned to the fasted level after 24 h. Plasma insulin concentration in chicks refed for 1 h was higher than in the fasted group. There was no significant change in plasma IGF-I concentration. 6. These results suggest that the FSR of breast muscle was more sensitive to refeeding than that of leg muscle which may be explained, in part, by differences in sensitivity to the change in circulating plasma insulin concentration after refeeding.     

Posttranslational processing of progonadotropin-releasing hormone (PROGnRH) in ewes

Gonadotropin-releasing hormone (GnRH) is released from hypothalamic neurons into the hypophyseal-portal blood system following enzymatic cleavage of the decapeptide from a large precursor (proGnRH) molecule. The purpose of this study was to determine whether the ability of GnRH-producing neurons to synthesize and/or process proGnRH differed during physiological states associated with a suppressed and enhanced release of GnRH in ewes. Tissues were collected from ovariectomized ewes (OVX, N=4), OVX-estradiol treated ewes (OVX-E, N=5), and ewes (n=7) slaughtered 5 d after parturition (PP). Following euthanasia and exsanguination, stalk-median eminence (SME), medial-basal hypothalamus (MBH) and preoptic areas (POA) were collected. Concentrations of GnRH and proGnRH were determined by radioimmunoassay using specific antisera. Concentrations of GnRH in the SME did not differ (P>.05) between OVX-E and OVX ewes, but both groups contained less (P<.05) GnRH than the SME from PP ewes (4.4 ± 0.7, 12.1 ± 3.8 vs 24.3 ± 5.1 fmol/mg tissue, respectively). Concentrations of proGnRH in SME mimicked those of GnRH and were less (P<.05) in OVX-E ewes than PP ewes, but were not different (P>.05) from those in OVX ewes (.34 ± .34 vs 3.76 ± 1.53 and 1.7 ± .78 fmol/mg, respectively). In the MBH, OVX-E ewes had greater (P<.05) concentrations of GnRH than PP ewes (0.76 ± 0.29 vs 0.24 ± 0.04 fmol/mg) and OVX ewes were intermediate (0.41 ± 0.13 fmol/mg). No differences (P>.05) in concentrations of GnRH in the POA were detected among groups. Concentrations of proGnRH in MBH and POA were not different (P>.05) among groups. In summary, proGnRH is present in the SME which contains nerve terminals of GnRH-producing cells. Although concentrations of proGnRH and GnRH in the SME and MBH were affected by physiological state, ratio's of the prohormone:mature decapeptide remained constant. Therefore, alterations in posttranslational processing of the prohormone leading to formation of the mature GnRH-decapeptide were not demonstrated.     

Effects of fasting and refeeding on the antioxidant system in cockerels and pullets

The effect of fasting and refeeding on total antioxidant status (TAS), glutathione peroxidase (GSH-Px) activity and concentration of some non-enzymatic antioxidant compounds was studied in cockerels and pullets. Blood was collected before and after 48-h fasting and 24 h after refeeding. In cockerels, fasting resulted in a significant decrease of TAS and uric acid concentration. After refeeding, the concentration of TAS remained significantly lower as compared to the control level. At the same time, blood plasma level of total lipids increased in comparison to the control and post-fasting values. In pullets, fasting resulted in a significant decrease of whole blood haemolysate GSH-Px activity and blood plasma concentrations of albumin and uric acid. Simultaneously, a significant increase in total lipids and cholesterol was obtained. In pullets, refeeding resulted in a further decrease of TAS to undetectable values, a significant decrease of blood plasma cholesterol, and a significant increase of GSH-Px in the whole blood haemolysate and in blood plasma uric acid content. The results indicate that fasting has a negative impact on the antioxidant defence system of the blood, which leads to a reduced resistance to oxidative stress in both cockerels and pullets. However, pullets seem to be more susceptible to fasting-provoked oxidative stress than cockerels.     

Effects of Estradiol and Progesterone on the Numbers and Distribution of Mastocytes in the Uterus of Ovariectomized Rats

The purpose of this study was to investigate the effects of estradiol(E)and progesterone(P)on mastocyte distribution in the uterus of ovariectomized rats.Thirty-five adult female rats were divided randomly into seven groups:one sham operated control group(SHAM);one ovariectomized group(OVX);three ovariectomized plus E treatment groups(OVX+E 20,100,or 500 μg/kg body weight·d);and two ovariectomized plus P groups(OVX+P 2 or 10 mg/kg body weight·d).Seven days after treatment,the contents of estradiol and progesterone in serum were detected by radioimmunoassay,and mastocytes in the uterus were stained by toluidine blue staining.Results were as following:① Compared to ovariectomized rat,the concent ration of estradiol in serum increased by 97.13 % in OVX+E 20(P0.05),204.84 % in OVX+E 100(P0.05),and 936.45 % in OVX + E 500 group(P0.05);the progesterone concent ration increased by 77.25 % in OVX+P 2(P0.05)and 235.25 %in OVX+P 10 group(P0.05).② Compared to ovariectomized rat,the number of mast cells in uteri decreased by 32.65% in OVX+E 20,64.50 % in OVX+E 100(P0.05),74.49 % in OVX+E 500(P0.05)and 70.67 % in OVX+P 10 groups(P0.05).However,the number of mast cells increased by 66.73% in OVX+P 2 group(P0.05)compared with OVX.The trend of mast cells number in the rat uterus was decreased gradually with the increase of estrogen or progesterone concent ration.The number of mast cells in ovariectomized rat uterus was affected by estrogen or progesterone.These results demonstrated that estrogen or progesterone directly affected the number of mast cells in the uterus of rat.