Opioidergic, dopaminergic and adrenergic regulation of LH secretion in prepubertal heifers

Studies have shown inhibitory effects of endogenous opioids on LH secretion in early post-natal heifers. However, it is not clear whether these effects change during the rest of the prepubertal period or whether the inhibitory influences on the GnRH neurones are direct or by way of other neuronal systems. Two experiments were performed in heifer calves to study the developmental patterns of opioidergic, dopaminergic and adrenergic regulation of LH and the possible interactions between opioids and dopaminergic and adrenergic neuronal systems, in the regulation of LH secretion. In Expt 1 four groups each of five heifer calves were used. Blood samples were taken every 15 min for 10 h and each calf received one of the following treatments as a single injection at 4, 14, 24, 36 and 48 weeks of age: (i) naloxone (opioid antagonist, 1 mg kg(-1), i. v.); (ii) sulpiride (dopamine D2 antagonist, 0.59 mg kg(-1), s.c.); (iii) naloxone and sulpiride combined; or (iv) vehicle (control group). Treatments began after the first blood sample was taken. The design of Expt 2 was similar; a separate group of heifer calves was assigned to receive one of the following treatments as a single injection at 4, 14, 24, 36 and 48 weeks of age: (i) naloxone; (ii) phenoxybenzamine (an alpha-adrenoreceptor blocker, 0.8 mg kg(-1), i. v.); (iii) naloxone and phenoxybenzamine; (iv) or vehicle. Results from Expt 1 showed that the maximum concentration of LH and the number of calves responding to treatments with an LH pulse was higher in the first hour after treatments at 36 and 48 weeks of age in the naloxone group compared with the control or sulpiride groups (P < 0.05). These values in the naloxone group also increased over time and were greatest at 48 weeks of age (P < 0.05). In heifers given naloxone + sulpiride treatment at 36 and 48 weeks of age, maximum concentrations of LH in the first hour after treatment did not differ from the naloxone and control groups. In Expt 2, at 36 and 48 weeks of age, treatment with naloxone with or without phenoxybenzamine resulted in higher concentrations of LH than in the controls (P < 0.05). No pulses were seen over the first hour of treatment at 36 and 48 weeks of age in heifers treated with phenoxybenzamine. The 10 h periods of blood sampling at 48 weeks of age revealed that phenoxybenzamine alone suppressed LH pulse frequency and mean serum concentrations of LH compared with the control group (P < 0.05). It was concluded that a strong or more acute inhibition of LH secretion by endogenous opioids developed in mid- to late prepubertal heifers, or alternatively, that removal of opioidergic inhibition at the GnRH neurone unmasked stimulatory inputs that were greater in heifers close to first ovulation. Since sulpiride appeared to negate in part the effects of naloxone on LH release, the suppressive effects of opioids could be exerted in part through the inhibition or blocking of a stimulatory dopaminergic system. alpha-Adrenergic neuronal systems have stimulatory effects on LH release, especially during the late prepubertal period, but do not appear to mediate opioidergic inhibition of LH secretion in prepubertal heifer calves.     

Luteinizing hormone secretion as influenced by age and estradiol in the prepubertal gilt

The aim of this study was to determine if there is an age related reduction in the sensitivity of the negative feedback action of 17β-estradiol (estradiol) on luteinizing hormone (LH) secretion in the prepubertal gilt. Ovariectomized gilts at 90 (n=12), 150 (n=11) or 210 (n=12) days of age received estradiol benzoate (EB) osmotic pump implants 6/group and the remaining animals received vehicle control (C) implants except for 150-day C (n=5) on Day 0. On Day 10 blood samples were collected every 15 min for 8h and serum LH and estradiol concentrations were measured. Serum estradiol concentrations averaged 5 ± 1, 5 ± 1 and 7 ± 2 pg/ml for the 90-, 150- and 210-day-old gilts implanted with estradiol, respectively, whereas, serum estradiol concentrations was undetectable in C gilts. Mean serum LH concentrations, basal LH concentrations and serum LH pulse amplitude were less in EB-treated gilts at all ages compared to control animals. In contrast, LH pulse frequency initially was less in EB-treated gilts but subsequently increased (P<0.04) with age (from 0.8 ± 0.2 at 90 days to 5.2 ± 0.2/8h at 210 days), and at 210 days of age the pulse frequency was similar to C gilts. These results demonstrate an age related reduction in the sensitivity to the negative feedback action of estradiol on LH secretion and support the idea that the gilt conforms to the gonadostat hypothesis.     

Absence of brain opioid peptide modulation of luteinizing hormone secretion in the prepubertal gilt

Three experiments were conducted to evaluate the role of endogenous opioid peptides (EOP) in modulating luteinizing hormone (LH) secretion in the prepubertal gilt. In Experiment I, 8 prepubertal (P) gilts, 160-170 days of age (puberty = 197 +/- 10 days), received either 1 (n = 2), 3 (n = 3), or 6 (n = 3) mg/kg BW of naloxone (NAL), an opiate antagonist, in saline i.v. Blood was collected by jugular vein cannula every 15 min for 2 h before and 2 h after NAL. All doses of NAL failed to alter serum LH concentrations. In Experiment II, 21 P gilts 160-170 days of age and 21 mature (M) gilts were ovariectomized (OVX). At the time of OVX, gilts were classified as prepubertal if their ovaries were devoid of corpora albicantia and corpora lutea. Three weeks after OVX, P and M gilts were injected twice daily for 10 days with either 0.85 mg/kg BW of progesterone (P4) or oil vehicle (V), resulting in the following groups: PP4 (n = 11), PV (n = 10), MP4 (n = 11), and MV (n = 10). All gilts received 1 mg/kg BW of NAL on the last day of treatment. Blood samples were collected via a jugular cannula every 15 min for 4 h before and 2 h after NAL treatment. NAL treatment resulted in an increase (p less than 0.05) in serum LH concentrations only in the MP4 gilts. In Experiment III, 15 OVX gilts 280 days of age were used. Ten of the 15 gilts were OVX prior to puberty at 160 days of age and were classified as chronologically mature (CM) at the time of treatment. The remaining 5 gilts were OVX after puberty, and were classified as sexually mature (SM) at the time of treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

Kinetics of follicle growth in the prepubertal gilt.

Follicular growth rates were determined by histological examination of ovaries of five prepubertal gilts following treatment with the stathmokinetic agent colchicine. One ovary from each of five gilts was removed surgically and then colchicine (n = 3) or saline (n = 2) was infused i.v. Precisely 2 h after treatment with colchicine, the remaining ovary was removed. Ovaries were processed for histological analyses and sectioned at 10 microns; every twentieth section was stained with hematoxylin and periodic acid-Schiffe's. Sections were viewed with a projection microscope and individual follicles were measured. Eight classes of follicles were established such that the number of granulosa cells per cross section doubled in each class. Diameters of follicles for each class were as follows: 1) less than 106 microns, 2) 106-148 microns, 3) 148-206 microns, 4) 206-287 microns, 5) 287-400 microns, 6) 400-657 microns, 7) 657-1480 microns, and 8) 1480-3130 microns. A layer of thecal cells was first seen in class 2 follicles, and 76% of class 3 follicles had a thecal layer. Oocyte diameter increased through the first four classes and reached a maximum diameter of approximately 110 microns. Almost all follicles greater than 400 microns had an antrum. Preantral follicles had a lower mitotic index and a higher mitotic time and class time than antral follicles. Growth rate increased with increasing size of follicles. Preantral follicles grew at a rate of 5.2 microns/day whereas antral follicles grew at 313 microns/day.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of restricted feeding and realimentation periods on compensatory growth performance and physiological characteristics of pigs

An experiment with 94 growing pigs was conducted to determine the effect of a feed restriction of 25% on performance, carcass quality, organ weight, blood hormone levels and some biochemical parameters. The experiment consisted of four periods of 21 days each. In the different periods animals (initial BW about 31 kg) were fed ad libitum (A) or restrictively (R), resulting at day 84 in Groups AAAA, AARA, RAAA and RARA. During Period I, the daily gain of restrictively fed pigs (Group R) was about 22% lower than from Group A (p < 0.01). During realimentation, compensatory growth was observed in Period II for Group RA, and in Period IV for Group RARA. No compensatory growth was observed for Group AARA, which was fed restrictively in Period III only (day 43 to 63). For the whole experiment (day 1 to 84), BW gain and feed conversion amounted to 830 g/d and 3.03 kg/kg, 798 g/d and 2.99 kg/kg, 813 g/d and 2.86 kg/kg, and 800 g/d and 2.78 kg/kg for Groups AAAA, AARA, RAAA and RARA, respectively. The decrease of liver and kidney weights as a result of restricted feeding was not significant and after three weeks of realimentation these differences almost disappeared. At day 3 after realimentation of restrictively fed pigs (Group RA) the growth hormone level was significantly increased, but at day 14 of realimentation this level turned out to be lower (p < 0.01) than in pigs fed ad libitum (Group AA). This was considered as a further indication of compensatory growth.     

Effect of restricted feeding and realimentation periods on compensatory growth performance and physiological characteristics of pigs

An experiment with 94 growing pigs was conducted to determine the effect of a feed restriction of 25% on performance, carcass quality, organ weight, blood hormone levels and some biochemical parameters. The experiment consisted of four periods of 21 days each. In the different periods animals (initial BW about 31 kg) were fed ad libitum (A) or restrictively (R), resulting at day 84 in Groups AAAA, AARA, RAAA and RARA. During Period I, the daily gain of restrictively fed pigs (Group R) was about 22% lower than from Group A (p < 0.01). During realimentation, compensatory growth was observed in Period II for Group RA, and in Period IV for Group RARA. No compensatory growth was observed for Group AARA, which was fed restrictively in Period III only (day 43 to 63). For the whole experiment (day 1 to 84), BW gain and feed conversion amounted to 830 g/d and 3.03 kg/kg, 798 g/d and 2.99 kg/kg, 813 g/d and 2.86 kg/kg, and 800 g/d and 2.78 kg/kg for Groups AAAA, AARA, RAAA and RARA, respectively. The decrease of liver and kidney weights as a result of restricted feeding was not significant and after three weeks of realimentation these differences almost disappeared. At day 3 after realimentation of restrictively fed pigs (Group RA) the growth hormone level was significantly increased, but at day 14 of realimentation this level turned out to be lower (p < 0.01) than in pigs fed ad libitum (Group AA). This was considered as a further indication of compensatory growth.     

Steroid control of gonadotrophin secretion in the orchidectomized dog

The effects of s.c. administration of oil, testosterone, 5 alpha-dihydrotestosterone, 5 alpha-androstane-3 alpha, 17 beta-diol and oestradiol-17 beta on plasma concentrations of LH and FSH were determined in 5 orchidectomized dogs. The dosages for the androgens and oestradiol-17 beta were 500 and 50 micrograms/kg body weight, respectively. Testosterone and oestradiol-17 beta significantly reduced plasma gonadotrophin concentrations, although the onset and duration of their suppressive effects differed. Dihydrotestosterone and oil had no effect on either gonadotrophin. Administration of androstanediol had no effect on plasma concentrations of LH but did cause a temporary and significant reduction in FSH. It is concluded that testosterone and oestradiol-17 beta are major regulators of gonadotrophin secretion in the male dog, but the 5 alpha-reduction of testosterone seems to play only a minor role in this control.     

Oestrogen and progesterone interactions in the control of gonadotrophin and prolactin secretion

Oestrogen and progesterone have marked effects on the secretion of the gonadotrophins and prolactin. During most of the oestrous or menstrual cycle the secretion of gonadotrophin is maintained at a relatively low level by the negative feedback of oestrogen and progesterone on the hypothalamic-pituitary system. The spontaneous ovulatory surge of gonadotrophin is produced by a positive feedback cascade. The cascade is initiated by an increase in the plasma concentration of oestradiol-17 beta which triggers a surge of luteinizing hormone releasing hormone (LHRH) and an increase in pituitary responsiveness to LHRH. The facilitatory action of oestrogen on pituitary responsiveness is reinforced by progesterone and the priming effect of LHRH. How oestrogen and progesterone exert their effects is not clear but the facilitatory effects of oestrogen take about 24 h, and the stimulation of LHRH release is produced by an indirect effect of oestradiol on neurons which are possibly opioid, dopaminergic or noradrenergic and which modulate the activity of LHRH neurons. In the rat, a spontaneous prolactin surge occurs at the same time as the spontaneous ovulatory gonadotrophin surge. The prolactin surge also appears to involve a positive feedback between the brain-pituitary system and the ovary. However, the mechanism of the prolactin surge is poorly understood mainly because the neural control of prolactin release appears to be mediated by prolactin inhibiting as well as releasing factors, and the precise role of these factors has not been established. The control of prolactin release is further complicated by the fact that oestradiol stimulates prolactin synthesis and release by a direct action on the prolactotrophes. Prolactin and gonadotrophin surges also occur simultaneously in several experimental steroid models. A theoretical model is proposed which could explain how oestrogen and progesterone trigger the simultaneous surge of LH and prolactin.     

Suppression of ovarian activity in the gilt and reversal by exogenous gonadotrophin administration

The aim of the current experiment was to study the regulation of follicle development in the pig using a potent GnRH agonist (GnRH-A) to initially suppress follicle development. Large-White hybrid gilts (n = 8) were treated during the luteal phase with GnRH-A. Four of these GnRH-A treated gilts and four control gilts were given a GnRH bolus on days 14 and 28 after GnRH-A administration or during the luteal phase in control gilts. Blood samples were collected for 10 h for FSH and LH, after which 1500 IU PMSG were administered and the ovaries and uteri recovered 72 h later. A further four GnRH-A treated gilts and four control gilts were slaughtered either 28 days after GnRH-A administration or during the luteal phase respectively, and all follicles > or = 1 mm diameter were dissected. The mean basal plasma FSH level was lower (P < 0.01) in GnRH-A treated than control gilts and showed no response to the GnRH challenge although levels increased (P < 0.01) in control gilts. The mean basal plasma LH levels were similar (P > 0.1) in GnRH-A treated and control gilts. Whilst in GnRH-A treated gilts plasma LH levels showed no response to the GnRH challenge, plasma LH levels were increased (P < 0.01) in control gilts. Pulsatile LH secretion was abolished in GnRH-A treated but not in control gilts. Plasma oestradiol levels were lower (P < 0.001) in GnRH-A treated gilts than in control gilts, but nevertheless both GnRH-A treated and control gilts responded to PMSG with increased plasma oestradiol levels. Treatment with GnRH-A reduced both the mean (2.1 vs. 2.7 mm; P < 0.01) and the maximal follicle diameter (4 vs. 6 mm) and reduced (P < 0.01) the total number of follicles > or = 2 mm diameter compared with control gilts. Administration of PMSG increased both mean follicle diameter (5.1 vs. 4.4 mm; P < 0.01) and maximal follicle diameter (7 vs. 9 mm) and caused a reduction (P < 0.001) in the total number of follicles > or = 2 mm diameter in both GnRH-A treated and control gilts. In summary, this study has demonstrated, for the first time in the pig, that the inhibition of follicle development as a result of pituitary down regulation/desensitisation can be reversed by exogenous gonadotrophin treatment. This model will be a powerful tool with which to investigate the precise regulation of follicle development in the pig.     

Effects of restricted feeding of prepubertal ewe lambs on growth performance and mammary gland development

The aim of this study was to determine the effects of restricted feeding before puberty on growth performance and mammary gland development in replacement ewe lambs. At weaning, 72 Dorset ewe lambs were assigned to one of the three diets: an ad libitum control diet with medium-quality forage (MQF; diet A-MQF); a restricted diet with the same forage as A, but less feed concentrate (diet R-MQF); or a high-quality forage (HQF) diet (diet F-HQF). The quantity of concentrate offered to the group R-MQF and F-HQF ewe lambs was adjusted to obtain 70% of the control ewe lambs' growth rate. The diets were offered for 75 days after weaning to cover the allometric phase of mammary gland development. During this period, average daily gain (ADG) was 223 and 229 g/day for groups R-MQF and F-HQF, respectively, compared to 305 g/day for group A-MQF (P < 0.0001). At the end of this period, 28 ewe lambs were slaughtered and their mammary gland was collected. Parenchymal fresh tissue weight tended to be higher for groups R-MQF and F-HQF compared to group A-MQF (P = 0.09). Stroma weight was greater (P < 0.05) for the group A-MQF ewe lambs than for those in the other treatments. Total DNA and total protein in parenchymal tissue tended to be greater for groups R-MQF and F-HQF (P = 0.09 and P = 0.07, respectively). Dry fat-free tissue was greater for groups R-MQF and F-HQF (P < 0.05). The remaining ewe lambs were fed the same haylage and barley diet until their first lambing. During this period, compensatory growth was observed. ADG was greater (P < 0.01) for groups R-MQF and F-HQF than for group A-MQF, and feed conversion was improved (P < 0.01) for groups R-MQF and F-HQF compared with the control, whereas the dry matter intake was similar for all groups. Live body weight, loin eye depth and backfat depth at breeding and around lambing were similar for all groups. The results of this study suggest that restricted feeding before puberty improves mammary gland development without compromising growth performance in ewe lambs.     

Opioidergic control of gonadotropin secretion in the female rabbit: divergent effects of morphine on secretion of follicle-stimulating hormone and luteinizing hormone

The nature of secretion of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) was followed in female rabbits on a daily basis from age 36 to 60 days by sequential 5-min blood sampling over 1- to 2-h periods each day. Both LH and FSH were found to be secreted in a pulsatile manner. The mean LH pulse amplitude over the 25 days was 0.95 +/- 0.32 ng/mL and for FSH it was 10.15 +/- 1.11 ng/mL. Mean plasma LH levels were significantly increased from 1.46 +/- 0.08 ng/mL in 36 to 42-day-old rabbits to 1.89 +/- 0.12 ng/mL in 43 to 50-day-old rabbits and remained elevated from 50 to 60 days. FSH levels during the same periods also rose significantly from 14.93 +/- 0.79 to 19.57 +/- 2.05 ng/mL. To examine the influence of endogenous opioid peptides on the release of LH and FSH in 36 to 60-day-old female rabbits, morphine sulfate at 0.2, 0.5, 2.0, and 5.0 mg/kg was administered subcutaneously after 30 min baseline sampling, and blood was taken for another 60-120 min. Morphine at all doses and at all ages inhibited the amplitude and frequency of LH pulses but had no effect on FSH secretion. To determine whether the effects of morphine on LH secretion could be reversed with naloxone, females aged 82-114 days were used. Naloxone administered 1 h after morphine reversed the inhibitory effects of morphine, whereas the simultaneous administration of naloxone with morphine had variable effects but seemed to delay the LH increase.(ABSTRACT TRUNCATED AT 250 WORDS)     

The influence of male contact on plasma cortisol concentrations in the prepubertal gilt

Large White X (Large White X Landrace) prepubertal gilts, 165 days of age, were fitted with indwelling venous catheters and housed in modified metabolism crates. After a period of acclimatization, frequent blood samples were taken at regular intervals before, during and after the 7 gilts were exposed to various degrees of contact with male pigs. The plasma samples were assayed for cortisol concentration using a competitive protein-binding radioassay. Significantly elevated concentrations of plasma cortisol (P less than 0.001) occurred only when full physical contact between the boar and the gilts was allowed. Boar exposure without full physical contact induced only minor changes in plasma cortisol concentrations of gilts. Plasma cortisol concentrations have been shown to constitute a reliable indicator of a stress response in pigs, and so the results of this study suggest that tactile stimulation from a male pig induces a stress response in the recipient prepubertal gilt. This stress response in the gilt may be involved in the stimulation of puberty onset by contact with a mature boar (i.e. the 'boar effect').     

Interactions between inhibin, oestradiol and progesterone in the control of gonadotrophin secretion in the ewe

Experiments were carried out to test the hypothesis that inhibin and oestradiol act synergistically to inhibit the secretion of FSH, to test for effects of progesterone, and to compare the FSH and LH responses to ovarian feedback. In Exp. 1, with 11 ovariectomized and 12 intact Romanov ewes during the anoestrous season, doses of oestradiol (administered by means of subcutaneous implants) that restored normal LH pulse frequencies were insufficient to restore normal concentrations of FSH. In Exp. 2, with 48 ovariectomized Welsh Mountain ewes during the breeding season, a factorial design with 4 ewes per cell was used to assess the responses in LH and FSH to 3 doses of oestradiol (s.c. implants) and 4 doses of bovine follicular fluid ('inhibin', 0.2-1.6 ml s.c. every 8 h). This was done initially in the absence of progesterone and then after 7 days of treatment with progesterone (s.c. implants). Analysis of variance revealed a significant synergistic interaction between oestradiol and inhibin on the plasma concentrations of FSH. Progesterone had little effect. In contrast, there was a significant synergistic interaction between oestradiol and progesterone on the concentrations of LH. 'Inhibin' also inhibited LH secretion but this effect was independent of the two steroids. We conclude that there are basic differences in the way that ovarian feedback acts to control the secretion of LH and FSH in the ewe. FSH secretion appears to be primarily controlled by the synergistic action of oestradiol and inhibin on the anterior pituitary gland, while the secretion of LH is inhibited during the follicular phase by an effect of oestrogen at pituitary level and during the luteal phase by the synergistic action of oestradiol and progesterone at the hypothalamic level. Inhibin, or another non-steroidal factor in follicular fluid, may also play a minor role in the control of LH secretion.     

Effects of restricted feeding of prepubertal ewe lambs on reproduction and lactation performances over two breeding seasons

The aim of this study was to determine the effects of restricted feeding before puberty on reproduction, lactation and offspring growth performance in replacement ewe lambs over two breeding seasons. At weaning, 41 Dorset ewe lambs were assigned to one of three diets: an ad libitum control diet with medium-quality forage (MQF; 13.3% crude protein (CP), 1.81 Mcal metabolizable energy per kg, 42.8% ADF; diet A-MQF); a restricted diet with the same forage as A but less feed concentrate (diet R-MQF); or a high-quality forage (HQF) diet (14.8% CP, 2.15 Mcal ME/kg, 34.7% ADF; diet F-HQF). The quantity of concentrate offered to the group R-MQF and F-HQF ewe lambs was adjusted to obtain 70% of the control ewe lambs' growth rate. The diets were offered for 75 days following weaning to cover the allometric phase of mammary gland development. Prepubertal restriction did not affect (P > 0.10) the gestation rate, number of lambs born or the body weight and body condition score of ewes at lambing or at the end of lactation. Ewes from groups R-MQF and F-HQF tended to produce more milk during their first lactation compared to those from group A-MQF (P = 0.07). During the second lactation, groups R-MQF and F-HQF had better standardized milk production than group A-MQF (P < 0.05), and group R-MQF produced more milk than group F-HQF (P < 0.05). Milk fat and protein content were not affected by treatments (P > 0.10) Fat and protein yield were affected by treatments only at the second lactation (P < 0.10 and P < 0.05, respectively). Lamb birth and weaning weights were not affected by prepubertal restriction of feeding in their mother (P > 0.10). However, the average daily gain of second breeding season lambs was higher for the R-MQF group than the F-HQF group (P < 0.05), and a similar trend was observed for total gain (P < 0.10). Restricted feeding before puberty does not impair future reproductive performance; however, it has a positive impact on lactation and on lambs' growth performance.     

Opioidergic and nutritional involvement in the control of luteinizing hormone secretion of postpartum Rasa Aragonesa ewes lambing in the mid-breeding season

The role of endogenous opioids and nutrition on the inhibition of luteinizing hormone (LH) secretion during the postpartum period was investigated in a Spanish breed of sheep lambing in the mid-late breeding season. Two groups of adult Rasa Aragonesa ewes housed in individual pens and lambing on 30 December were fed during the suckling period to provide maintenance requirements and the production of 1.1 (M; n=8) or 0.55 (L; n=8) kg of milk per day. On days 10, 20 and 30 after lambing, the effect of a treatment with the opiate receptor antagonist naloxone (1 mg/kg at four hourly intervals) on LH secretion was assessed in half of the ewes of each group, the remaining females receiving four saline injections. After weaning, animals were fed to provide requirements for maintenance of liveweight. Blood samples were collected twice a week from day 20 postpartum until the end of March, and assayed for progesterone and prolactin. Although underfed ewes showed significantly lower mean plasma concentrations during the control period on day 20 postpartum, nutrition did not seem to modify LH secretion before naloxone or saline injections. Moreover, no differences between nutritional groups in the response to naloxone injections on pattern of LH secretion were found. In fact, naloxone treatment induced an increase of mean LH concentrations on days 10, 20 and 30 postpartum (at least, P<0.05), of LH pulse frequency on days 20 and 30 (P<0.05), and of LH pulse amplitude on days 10 and 20 (P<0.05). Underfed ewes during the postpartum period showed a slower decline in plasma prolactin levels, with significant differences on days 29, 36 and 39 after lambing (P<0.05). Only 3 M ewes ovulated before the onset of the seasonal anoestrus period. It is concluded that endogenous opioids are involved in the inhibition of LH secretion during the early suckling period of a reduced seasonality breed of sheep without any influence of nutrition on the response to naloxone treatment; however, ewes underfed before weaning failed to reactivate their cyclicity prior to the onset of the seasonal anoestrus.     

Opioidergic control of luteinizing hormone secretion in the female rabbit: influence of age on the response to naloxone

To examine the role of opioid neurons on luteinizing hormone (LH) secretion in the female rabbit, we determined LH release at timed intervals after naloxone administration to rabbits aged 25-150 days. The LH response to naloxone (10 mg/kg) was not significantly elevated until day 43 when LH rose 76-113% above basal levels at 40-80 min. In 56-day-old females the corresponding increase was 160% at 15 min and in 65- to 67-day-olds it was 154%. From 70 to 80 days of age the LH response was blunted and no significant elevations could be elicited. By contrast, naloxone-induced LH increases were again evident when rabbits were older than 100 days. At all ages no significant change in FSH concentrations was observed. In the adult females, naloxone at 2.5, 5, and 10 mg/kg caused increases in LH secretion which occasionally were high enough to induce ovulation as exemplified by elevated serum progesterone 4 days later. These data suggest that opioid peptides may be involved in the prepubertal rise in LH and in the normal inhibition of adult secretion in the female rabbit.     

Opioidergic control of luteinizing hormone release in the female rabbit: influence of ovariectomy and steroid replacement on pulsatile secretion

The influence of ovariectomy and steroid replacement on naloxone-induced changes in pulsatile secretion of luteinizing hormone (LH) in the female rabbit was examined. Blood samples were taken every 5 min through an indwelling catheter in the rabbit ear artery, and plasma was stored until assayed for LH by established radioimmunoassay procedures. In the intact animal, saline injection had no effect on LH secretion. Although naloxone (10 mg/kg) caused a 7-fold increase in mean LH pulse amplitude by 30 min after injection, this increase was not statistically significant because 5 of 11 animals did not respond. In animals ovariectomized 48 h previously, naloxone significantly increased LH concentration by 194% at 23 min after injection. When long-term ovariectomized rabbits were treated with estradiol benzoate and then were given naloxone, no significant increase in LH was observed, although many animals did respond. Treatment of long-term ovariectomized rabbits with 1 microgram estradiol benzoate and 100 micrograms progesterone or 1 mg testosterone propionate on Days 1 and 3 and naloxone on Day 4 resulted in a significant increase in LH 19-24 min later. Although there was an increase in pulse amplitude, no change was detected in pulse frequency after naloxone. These data suggest that the hypothesis of steroid-opioid coupling in the control of LH secretion is not applicable to the female rabbit.     

Metabolic adaptations of liver mitochondria during restricted feeding schedules

Food anticipatory activity (FAA) is an output of the food-entrained oscillator (FEO), a conspicuous biological clock that expresses when experimental animals are under a restricted food schedule (RFS). We have shown that the liver is entrained by RFS and exhibits an anticipatory response before meal time in its oxidative and energetic state. The present study was designed to determine the mitochondrial oxidative and phosphorylating capacity in the liver of rats under RFS to further support the biochemical anticipatory role that this organ plays during the food entrainment (9). Metabolic and functional parameters of liver mitochondria were characterized before (0800 h), during (1100 h), and after (1400 h) FAA. The main results were as follows. First, there was an enhancement during FAA (1100 h) in 1) oxidative capacity (site I of the electron transport chain), 2) phosphorylating ability (estimated by ATP synthesis), and 3) activities of NADH shuttles. Second, after rats were fed (1400 h), the phosphorylating capacity remained high, but this was not the case for the respiratory control ratio for site I. Finally, in the three experimental conditions before, during, and after FAA, an increment was detected in the H(+) electrochemical potential, due to an elevation in mitochondrial membrane potential, and in mitochondrial yield. Most of the changes in mitochondrial properties related to RFS were also present when results were compared with those from the 24-h fasted group. In conclusion, the results support the notion that a distinctive rheostatic state is installed in the metabolic activity of the liver when FEO is being expressed.     

Interactions between serotoninergic and aminoacidergic pathways in the control of PRL secretion in prepubertal male rats

Prolactin secretion is controlled by the hypothalamus through different neurotransmitters which interact with multiple receptor subtypes. The discovery of different families of receptors for serotonin (5-HT₁-5-HT₇) and excitatory aminoacids (NMDA,KA,AMPA and metabotropic receptors) ilustrates the complexity of this regulation. Moreover, in the rat the role of different neurotransmitters changes during pubertal development. Present experiments were carried out to analyse the interactions between AMPA and serotoninergic receptors in the control of prolactin secretion in prepubertal male rats. For this purpose, 16 and 23-day old male rats were treated with 5-hydroxytryptophan (5-HTP, precursor of serotonin synthesis) plus fluoxetine (blocker of serotonin reuptake), 8-OH-DPAT (agonist of 5-HT_1A receptors), DOI and α-Me-5-HT (agonists of 5-HT₂ receptors), 1-phenylbiguanide (agonist of 5-HT₃ receptors) alone or in combination with AMPA (agonist of AMPA receptors). The results obtained indicate that: (a) activation of 5-HT_1A receptors stimulated PRL secretion on day 16 and inhibited it on day 23; activation of 5-HT₂ receptors stimulated PRL secretion on days 16 and 23, whereas activation of 5-HT₃ receptors inhibited PRL release only on day 23; (b) activation of AMPA receptors inhibited PRL secretion on day 23, but not on day 16 and (c) a cross-talk is apparent between 5-HT₂ and AMPA receptors in the regulation of PRL secretion, the stimulatory effect of DOI being blocked by AMPA.