Reproduction

Molteno, A J; Kalló, I; Bennett, N C; King, J A; Coen, C W

doi: 10.1530/rep.1.00048pmid: 15056766

The gonadotrophin-releasing hormone (GnRH) system in female Damaraland mole-rats, Cryptomys damarensis, has been investigated to map the distribution of GnRH-immunoreactive (GnRH-IR) structures in the brain of this species and to assess whether changes in this system may mediate the inhibitory effect of social cues on fertility. The distribution of GnRH-IR cell bodies and fibres was similar to that of other mammals, forming a loose continuum along a septo-preoptico-infundibular pathway. GnRH-IR cell bodies were more abundant in the vicinity of the organum vasculosum of the lamina terminalis than in the medial basal hypothalamus. GnRH-IR cells and fibres were also found in the subfornical organ. The cell bodies were typically unipolar or bipolar. No differences were found in the morphology or size of the cell bodies or in the number of cells between non-reproductive females and reproductive females living together in a colony. However, GnRH concentrations, measured in the brain by radioimmunoassay, were significantly higher in non-reproductive females than in reproductive females; this finding was complemented by the reduced immunoreactivity for GnRH in the median eminence and proximal pituitary stalk of reproductive females. In contrast, the concentrations of GnRH measured by radioimmunoassay in non-reproductive and reproductive males did not differ. These results are consistent with the hypothesis that GnRH release is inhibited in the non-reproductive females but not in the non-reproductive males of this species.

Haughian, J M; Ginther, O J; Kot, K; Wiltbank, M C

doi: 10.1530/rep.1.00030pmid: 15056767

Preovulatory LH and FSH surges and the subsequent periovulatory FSH surge were studied in heifers treated with a single injection of GnRH (100 μg, n = 6) or saline (n = 7). Blood samples were collected every hour from 6 h before treatment until 12 h after the largest follicle reached ≥8.5 mm (expected beginning of follicular deviation). The GnRH-induced preovulatory LH and FSH surges were higher at the peak and shorter in duration than in controls, but the area under the curve was not different between groups. The profiles of the preovulatory LH and FSH surges were similar within each treatment group, suggesting that the two surges involved a common GnRH-dependent mechanism. Concentrations of FSH in controls at the nadir before the preovulatory surge and at the beginning and end of the periovulatory surge were not significantly different among the three nadirs. A relationship between variability in the periovulatory FSH surge and number of 5.0 mm follicles was shown by lower FSH concentrations during 12–48 h after the beginning of the surge in heifers with more follicles (11.0 ± 1.0 follicles (mean±s.e.m.) n = 7) than in heifers with fewer follicles (5.7 ± 0.4, n = 6). This result was attributed to increased FSH suppression from increased numbers of follicles reaching 5.0 mm. Grouping of heifers into those with longer vs shorter intervals from a 4.5 mm to an 8.5 mm largest follicle did not disclose any relationship between length of the interval and FSH characteristics (e.g. profile of surge, area under curve, FSH concentrations at specific events). The hypothesis of a relationship between variation in the periovulatory FSH surge and variation in follicular dynamics was supported for the number of 5.0 mm follicles but not for the hour the largest follicle reached 8.5 mm. Thus, the expected time of follicle deviation was not altered by the extensive variation in the wave-stimulating FSH surge.

Amstalden, M; Zieba, D A; Garcia, M R; Stanko, R L; Welsh, T H; Hansel, W H; Williams, G L

doi: 10.1530/rep.1.00022pmid: 15056768

Experiments were performed to test the hypothesis that lamprey GnRH-III (lGnRH-III) selectively releases FSH. Primary cultures of bovine adenohypophyseal cells were treated with mammalian GnRH (mGnRH) and lGnRH-III (10−9, 10−8, 10−7 and 10−6 M) or control media in Experiment 1. All doses of mGnRH and the two highest doses of lGnRH-III stimulated (P < 0.001) a non-selective release of LH and FSH. In Experiments 2–4, Latin Square designs were utilized in vivo to examine whether physiological and hormonal milieu regulate putative selective effects of lGnRH-III. In Experiments 2 and 3, ovariectomized cows with basal levels of estradiol only (Experiment 2) or in combination with luteal phase levels of progester-one (Experiment 3) were injected with mGnRH and lGnRH-III (0.055, 0.11, 0.165 and 1.1 μg/kg body weight (BW) and saline. All doses of mGnRH released (P < 0.001) LH and FSH, but only the highest dose of lGnRH-III stimulated (P < 0.001) a non-selective release of both LH and FSH (Experiment 3). For Experiments 4A and 4B, intact, mid-luteal phase cows were injected with mGnRH and lGnRH-III (1.1 μg/kg BW; Experiment 4A), lGnRH-III (1.1 and 4.4 μg/kg BW; Experiment 4B) and saline. As before, mGnRH released (P < 0.001) both LH and FSH at all doses. In contrast, lGnRH-III at the highest dose released (P < 0.001) LH but not FSH. These findings suggest that lGnRH-III may act as a weak competitor for the mGnRH receptor and do not support the hypothesis that it selectively releases FSH in cattle.

Mondal, M; Prakash, B S

doi: 10.1530/rep.1.00023pmid: 15056769

To investigate the effects of long-term GH-releasing factor (GRF) administration on the patterns of GH and LH secretion in growing female Murrah buffalo (Bubalus bubalis) calves, 12 buffaloes of 6–8 months of age were divided into two groups (treatment and control groups) of six each in such a way that average body weight between the groups did not differ significantly (P > 0.05). Both the groups were administered i.v. with either synthetic bovine GRF (bGRF(1–44)-NH2) at 10 μg/100 kg body weight (treatment group) or an equal volume of normal saline (control group) at intervals of 15 days until 18 injections had been completed (9 months). Blood samples collected prior to and after the first and last injection of GRF at −60, −45, −30, −15, −10, −5 min and +5, +10, +15, +30 min, and thereafter at intervals of 15 min up to 8 h post-injection, were assayed for plasma GH and LH. Plasma progesterone was also estimated in twice-a-week samples to assess whether either group had begun ovarian cyclicity. The body weight of all animals was recorded twice a week. In all animals, a peak of GH was recorded within 5–20 min and 5–30 min after the first and last GRF injections and post-injection mean values for plasma GH were significantly (P < 0.01) higher compared with the control group of animals. Although peak GH values after the first and last GRF injection did not differ (P > 0.05), GH levels were maintained at a higher level for a longer time after the last GRF injection compared with the first (240 vs 150 min). The area under the GH response curve after the last GRF injection was found to be significantly (P < 0.01) higher than after the first injection (9344 ± 99.7 vs 7763 ± 112.4 ng/ml × min). The mean post-injection plasma LH levels of the treatment group were significantly (P < 0.01) higher after both the first and last GRF injections than in the control group of animals. Interestingly, compared with the first GRF injection, the pre-injection plasma LH level was found to be significantly higher (P < 0.01) at the last injection. The plasma LH concentrations around the last injection of GRF were significantly higher (P < 0.01) than those recorded at the time of the first injection in treated buffaloes. Correspondingly, the plasma LH concentrations in controls were also higher (P < 0.01) around the last injection of GRF vis-à-vis the first injection. The hormone concentration exhibited a higher pulsatility with greater amplitude after the last injection as compared with that recorded after the first injection. Although pulses of LH were also recorded in controls following the last injection, these were fewer and of lower magnitude than those seen in treated animals. No animal from either group reached puberty. GRF-treated buffaloes attained higher (P < 0.001) body weight than the controls. In conclusion, long-term administration of GRF induces and even enhances GH release without any sign of refractoriness, and significantly increases plasma LH also. Hence, long-term treatment with GRF may be used to maintain a sustained increased level of plasma GH in buffaloes and it may assist the animals of this species to grow faster.

McGlothlin, J A; Lester, G D; Hansen, P J; Thomas, M; Pablo, L; Hawkins, D L; LeBlanc, M M

doi: 10.1530/rep.1.00021pmid: 15056770

An experimental model of ascending placentitis was developed in the mare to characterize the uterine myoelectrical pattern in late gestation and determine how ascending placentitis altered this pattern. In experiment 1, myometrial electrical activity was analyzed during the early morning, late morning and evening hours in four mares in the last 15 days of gestation to identify patterns of activity. In experiment 2, nine mares received intra-cervical inoculations of Streptococcus equi subspecies zooepidemicus. Myoelectrical activity in the early morning and evening hours in these mares was compared with four control mares. In experiment 1, the number of spike burst clusters >30 s was greater in the evening than in the late morning hours (P < 0.04). Spike burst activity (number × duration) of mares in experiment 1 was similar during day and night recordings until the last 6 days of gestation when it gradually increased each evening until parturition (P < 0.05). In experiment 2, control mares experienced a gradual increase in the number of small spike burst clusters in the last 6 days (P = 0.008) and an increase in large and small spike burst clusters in the evening hours in the last 4 days of gestation (P = 0.03). Mares with experimentally induced placentitis never exhibited a rise in spike burst clusters but had an increase in the mean duration and activity index of large spike burst clusters in the 4 days before parturition (P < 0.04). In conclusion, control mares had a progressive, reversible rise in myoelectrical activity at night in the week preceding parturition. This was not observed in mares with experimentally induced placentitis. They exhibited an increase in the intensity and duration of large spike burst clusters possibly in response to local inflammation.

Allen, W R; Wilsher, Sandra; Tiplady, Clare; Butterfield, R M

doi: 10.1530/rep.1.00024pmid: 15056771

The growth parameters exhibited by seven Thoroughbred (Tb) foals that had experienced a ‘restricted’ in utero existence following transfer as embryos to the uteri of smaller Pony (P) mares (Tb-in-P) and, conversely, six P foals that experienced a ‘luxurious’ in utero existence after transfer to larger Tb mares (P-in-Tb), were compared from birth to 3 years of age with those exhibited by six normal Tb-in-Tb and six P-in-P foals conceived by within-breed artificial insemination. Bodyweight, height at the withers, girth, poll-to-nose length, crown–rump length and three foreleg longbone measurements were made at regular intervals. At birth, an approximate 15% reduction or increase in parameters was observed in the Tb-in-P and P-in-Tb respectively, which declined to 5% by 3 years of age. Growth post partum was affected by restricted or enhanced growth in utero. In the first 6 months post partum, growth rate was enhanced in the previously restricted Tb-in-P foals and curbed in the previously enhanced P-in-Tb foals compared with their respective controls. Overall, the similarity of the responses of the offspring to both ‘restriction’ and ‘luxury’ in utero ensured that no major changes to conformation resulted from either treatment. Thus, the Thoroughbreds carried by the Pony mares were merely scaled down versions of the Tb-in-Tb controls while the Ponies carried by the Thoroughbred mares were scaled up versions of the P-in-P controls.

Battersby, S; Critchley, H O D; de Brum-Fernandes, A J; Jabbour, H N

doi: 10.1530/rep.1.00038pmid: 15056772

Prostacyclin (PGI2) synthesis and function in the human uterus has been implicated in the regulation of the process of normal and dysfunctional menstruation. PGI2 synthesis is elevated during normal menstruation and is also associated with blood loss in women who suffer from heavy menses. This study was designed to outline further the role of PGI2 in menstruation by investigating the temporal pattern and site of expression of prostaglandin I synthase (PGIS) and the prostacyclin receptor (IP receptor) in the non-pregnant human endometrium across the menstrual cycle. Quantitative RT-PCR demonstrated increased expression of PGIS and IP receptor during the menstrual phase of the cycle compared with all other phases (P < 0.05). Furthermore, PGIS and IP receptor were localised to the glandular epithelium, stromal and endothelial cells in the basal and functional layers of the endometrium. Functionality of the IP receptor in the human endometrium was assessed by measuring cAMP generation following treatment with 100 nmol l−1 of the PGI2 analogue, iloprost. cAMP generation was significantly higher in endometrial tissue collected during the proliferative compared with the secretory phase of the menstrual cycle (P < 0.05).In conclusion, this study has confirmed increased expression and signalling of PGIS and IP receptor during the menstrual phase and outlines a potential autocrine/paracrine role for PGI2 on several cellular compartments in the endometrium including the endothelium. This may underscore a pivotal role for PGI2 receptor signalling in normal and dysfunctional menstruation.

Henson, M C; Swan, K F; Edwards, D E; Hoyle, G W; Purcell, J; Castracane, V D

doi: 10.1530/rep.1.00037pmid: 15056773

Leptin produced by both adipose tissue and the placental trophoblast, has been proposed to regulate numerous aspects of human conceptus development. Although recent animal studies have suggested an additional role for the polypeptide in fetal lung maturation, no evidence has been reported in primates. Therefore, we employed the baboon (Papio sp.), a well-characterized primate model for human pregnancy, to determine the presence and ontogeny of leptin receptor in fetal lung with advancing gestation. Lungs were collected from fetal baboons, early in gestation (days 58–62, n = 4), at mid gestation (days 98–102, n = 4), and late in gestation (days 158–165, n = 4) (term 184 days). mRNA transcripts for leptin (LEP) and both long and short intracellular domain isoforms of the leptin receptor (LEP-RL and LEP-RS) were assessed by RT-PCR. leptin receptor protein was evaluated by immunoblotting and cell types expressing leptin receptor were identified in late pregnancy by immunohistochemistry. Fetal serum leptin concentrations, determined by RIA, remained relatively unchanged at 5.7 ± 1.1 ng/ml (mean ± s.e.m.) in mid pregnancy and 8.4 ± 3.0 ng/ml in late pregnancy (P > 0.05). Although leptin were detectable in fetal lung, no changes in transcript abundance were apparent with advancing gestation. However, transcripts for both LEP-RL and LEP-RS receptor isoforms increased several-fold (P < 0.05) in fetal lung between mid and late gestation, while leptin receptor protein was detectable only in late pregnancy. leptin receptor was localized in distal pulmonary epithelial cells, including type II pneumocytes. In conclusion, leptin is present in the fetal baboon and its receptor is enhanced during late gestation in cells responsible for the synthesis of pulmonary surfactant. Collectively, these and past findings may suggest a modulatory role for the polypeptide in pulmonary development and/or may identify leptin receptor as a physiological marker of primate fetal lung maturity.