The aim of the study was to investigate the effects of chronic nandrolone decanoate treatment and/or swimming training on immunohistomorphometric parameters on rat pituitary gonadotropic cells. Male Wistar albino rats, 10 weeks old, were classified into four groups: control (T−N−), nandrolone (T−N+), swimming training (T+N−), and swimming training with nandrolone (T+N+). The T+ groups swam for 4 weeks, 1 h/day, 5 days/week. The N+ groups received nandrolone decanoate (20 mg/kg) once per week for 4 weeks. Pituitary tissue sections were processed and stained for immunohistochemical analysis and immunofluorescence. The volume density of luteinizing hormone (LH) cells was decreased by 48% in T−N+ and for 35% in the T+N+ group. The volume density of follicle-stimulating hormone (FSH) cells was decreased by 39% in T−N+ and for 30% in T+N+ compared to the control. Nandrolone alone, or combined with swimming training, decreased the number of LH/FSH cells compared to the control. The levels of the immunofluorescent signal of LH/FSH cells were increased in all experimental groups. Nandrolone alone decreased the serum level of LH by 17%, whereas swimming training alone increased FSH levels by 11% compared to the control. Serum levels of testosterone were increased in all experimental groups. Nandrolone alone, or combined with swimming training, decreased immunohistomorphometric parameters of gonadotropic cells, whereas the levels of immunofluorescent signal were increased.

Gonadotropins are the key regulators of ovarian follicles development. They are applied in therapeutic practice in assisted reproductive technology clinics. In the present review we discuss the basic gonadotropic hormones – recombinant human follicle-stimulating hormone, its derivatives, luteinizing hormone and gonadotropin serum of pregnant mares, their origin, and application in ovarian follicle systems in in vitro culture systems.

Follicle stimulating hormone (FSH) and luteinizing hormone (LH) are commonly added to maturation media to improve cumulus expansion known to be a predictor of oocyte maturation. Therefore, effects of various concentrations of FSH (1000 ng/ml), LH (1000 ng/ml) and FSH + LH (1000 ng/ml each) in comparison with control (without FSH + LH) cultured oocytes were investigated. FSH and LH (1000 ng/ml each) induced significantly more cumulus expansion and polar body numbers, as compared with control and treatments of 1000 ng/ml FSH and 1000 ng/ml LH alone. Expression of FSH receptor (r), LHr and Cx43 mRNAs was determined by real-time PCR in cumulus–oocyte complexes (COCs) and denuded oocytes at different maturation times. Expression of all three genes was higher in COCs compared with denuded oocytes, confirming the importance of cumulus cells in oocyte maturation. FSHr and connexin 43 (Cx43) mRNA abundance in both COCs and denuded oocytes was highest at 0–6 h of maturation and decreased subsequently. However, LHr mRNA abundance increased from 6 h up to 24 h of maturation. The study concluded that FSH, LH receptors and Cx43 gene expression regulation is an index related to oocyte maturation.

Three experiments (EXP) were conducted to determine the role of insulin-like growth factor-I (IGF-I) in the control of growth hormone (GH) and LH secretion. In EXP I, prepuberal gilts, 65 ± 6 kg body weight and 140 days of age received intracerebroventricular (ICV) injections of saline (n = 4), 25 μg (n = 4) or 75 μg (n = 4) IGF-I and jugular blood samples were collected. In EXP II, anterior pituitary cells in culture collected from 150-day-old prepuberal gilts (n = 6) were challenged with 0.1, 10 or 1000 nM [Ala15]-h growth hormone-releasing hormone-(1-29)NH2 (GHRH), or 0.01, 0.1, 1, 10, 30 nM IGF-I individually or in combinations with 1000 nM GHRH. Secreted GH was measured at 4 and 24 h after treatment. In EXP III, anterior pituitary cells in culture collected from 150-day-old barrows (n = 5) were challenged with 10, 100 or 1000 nM gonadotropin-releasing hormone (GnRH) or 0.01, 0.1, 1, 10, 30 nM IGF-I individually or in combinations with 100 nM GnRH. Secreted LH was measured at 4 h after treatment. In EXP I, serum GH and LH concentrations were unaffected by ICV IGF-I treatment. In EXP II, relative to control all doses of GHRH increased (P < 0.01) GH secretion. Only 1, 10, 30 nM IGF-I enhanced (P < 0.02) basal GH secretion at 4 h, whereas by 24 h all doses except for 30 nM IGF-I suppressed (P < 0.02) basal GH secretion compared to control wells. All doses of IGF-I in combination with 1000 nM GHRH increased (P < 0.04) the GH response to GHRH compared to GHRH alone at 4 h, whereas by 24 h all doses of IGF-I suppressed (P < 0.04) the GH response to GHRH. In EXP III, all doses of IGF-I increased (P < 0.01) basal LH levels while the LH response to GnRH was unaffected by IGF-I (P > 0.1). In conclusion, under these experimental conditions the results suggest that the pituitary is the putative site for IGF-I modulation of GH and LH secretion. Further examination of the role of IGF-I on GH and LH secretion is needed to understand the inhibitory and stimulatory action of IGF-I on GH and LH secretion.

Successfully using artificial insemination (AI) is defined as getting cows pregnant when the farmer wants them in-calf and making the best use of appropriate genetic potential. Over the past 30 to 50 years, the percentage of animals in oestrus that stand-to-be-mounted (STBM) has declined from 80% to 50%, and the duration of STBM from 15 h to 5 h; both in parallel with a reduction in first-service-pregnancy-rate from 70% to 40%. Meanwhile, the incidence of lameness and mastitis has not decreased; and it takes more than an extra 40 and 18 days, respectively, to get a lame or mastitic cow in-calf compared to healthy herd-mates. The intensity of oestrus is 50% lower in severely lame cows, and fewer lame cows ovulate. Luteal phase milk progesterone concentrations are also 50% lower in lame cows, and follicular phase oestradiol is also lower in non-ovulating lame cows compared to ovulating animals. Furthermore, lame cows that do not ovulate do not have an LH surge, and the LH pulse frequency in their late follicular phase is lower (0.53 v. 0.76 pulses/h). Thus, we suggest that the stress of lameness reduces LH pulsatility required to drive oestradiol production by the dominant follicle. The consequent low oestradiol results in less-intense oestrus behaviour and failure to initiate an LH surge; hence there is no ovulation. A series of experimental studies substantiate our hypothesis that events activating the hypothalamus–pituitary–adrenal axis interfere at both the hypothalamus and the pituitary level to disrupt LH and oestradiol secretion, and thus the expression of oestrus behaviour. Our inability to keep stress at a minimum by appropriately feeding and housing high-production cows is leading to a failure to meet genetic potential for yield and fertility. We must provide realistic solutions soon, if we want to successfully use AI to maintain a sustainable dairy industry for the future.

The effects of body condition score (BCS) of 2·0 or 3·0 units at calving (low v. moderate), post-partum energy intake at 0·6 or 1·0 MJ metabolizable energy (ME) per day per kg M0·75 (low v. high) and unrestricted or restricted (once daily) suckling on the ability of cows to ovulate were studied in a 2 ✕ 2 ✕ 2 factorial design with each treatment replicated eight times. Calf isolation and restricted suckling were imposed shortly after selection of the first dominant follicle (DF) to emerge after day 21 post partum. The episodic release of LH (sampled at 15-min intervals for 10 h) was determined 48 h before and 48 h after the day calf isolation and restricted suckling commenced. Additional blood samples were collected weekly for plasma insulin determination. The mean interval from calving to first ovulation was shorter for cows in moderate than low BCS at calving (47·8 v. 57·1 days, s.e.d. = 4·50, P < 0·05), and for cows suckling once daily than for those with unrestricted suckling (42·9 v. 62·0 days, s.e.d. = 4·50, P < 0·001). Post-partum nutrition did not affect this interval. Mean LH pulse frequency prior to the start of restricted suckling was higher for cows of moderate than low BCS at calving (3·2 v. 1·6 pulses per 10 h, s.e.d. = 0·60, P < 0·05). Subsequently, LH pulse frequency was higher for cows suckling once daily than for those with unrestricted suckling (4·0 v. 2·2 pulses per 10 h, s.e.d. = 0·82, P < 0·05). More cows in moderate than low BCS ovulated the first DF to emerge after day 21 post partum (within 4 to 6 days) in response to restricted suckling (69 v. 25%, P < 0·05). LH pulse frequency prior to restricted suckling increased (P < 0·05) with plasma insulin concentration (categorized as low, < 5; moderate, 5 to 8; and high, >8 mIU per l). There were indications of interactions between suckling treatment and BCS (P < 0·08), and suckling treatment and plasma insulin concentration (P < 0·06), on LH pulse frequency, which suggested that calf restriction could alleviate the suppressive effects of under nutrition on episodic LH release. Amongst cows suckling once daily, the non-ovulating animals had fewer LH pulses prior to restricted suckling and smaller, slower growing DF, indicating an inability of the DF to respond to increased LH pulse frequency following calf restriction. Cows of moderate BCS, particularly those with moderate to high levels of plasma insulin (³ 5 mIU per l), responded favourably to restricted suckling. In contrast, excessively thin cows with low plasma insulin concentrations (<5 mIU per l), that had most to gain from restricted suckling, responded poorly.

Thirty-two post-partum Mediterranean river buffalo cows were continuously observed for signs of oestrus from September to December with the aid of two vasectomized bulls. Symptoms of oestrus among female Mediterranean buffaloes are weak, therefore oestrus was assessed based on bull behaviour (following and licking a cow and trying to mount her). Oestrus was divided into three phases based on the bull behaviour assessment. Status of the uterus and ovarian follicles were checked rectally every day for each cow which was detected by a teaser bull to be in oestrus. All cows in oestrus were bred twice by artificial insemination (AI), the first at ovulation and the second (using a different bull) 22 h later.

The average duration of interest shown by the bull towards a cow (from the very first to the last sign of interest) was 68 h and the average length of the three phases was: phase 1 = 21 (s.d. 29) h, phase II = 32 (s.d. 24) h and phase III = 15 (s.d. 15) h. Fifteen buffaloes were bled during oestrus, and LH profiles were determined. No differences were evident among oestruses followed by pregnancy (no. = 18) and the others (no. = 26) for the variables describing behavioural events. Neither intensity of the bull courtship, presence or clarity of mucous discharge, or housing system affected the success of AI. The only differences between pregnant and non-pregnant cows were in the timing between the LH peak and the end of phase II (2·4 v. 14·7 h, P < 0·001), end of phase III (22 v. 40 h, P > 0·05) and ovulation (25 v. 46 h, P < 0·05). Successful pregnancies occurred 34 (s.d. 14) h after the end of phase II. The endocrinology and behavioural patterns of buffalo reproduction need further research to clarify the reasons for non-optimal pregnancy rates after AI.

The role of endogenous opioids and the dopaminergic system on the inhibition of luteinizing hormone (LH) secretion during early and late anoestrus, together with its modulation by the plane of nutrition were investigated in ewes with a short anoestrous season. In early anoestrus (22 March; day 0), two groups of ovariectomized, oestradiol-treated adult Rasa Aragonesa ewes, maintained under natural photoperiod at 41°N, were given enough food to provide 1·4 × (high; H; no. = 6) or 0·5 × (low; L; no. = 6) energy requirements for maintenance. The effects of administration of the opiate receptor antagonist naloxone (1 mg/kg at four 1-h intervals) (day 15) and of the dopaminergic2 receptor antagonist pimozide (0·08 mg/kg) (day 21) on LH secretion were assessed. A second experiment was carried out in late anoestrus (21 June) using the same protocol. A significant increase in LH pulse frequency after naloxone treatment for both H and L groups was detected in late anoestrus. Number ofLH pulses after naloxone injections in early anoestrus also increased in H (P < 0·05) and L ewes (P = 0·08). The effect of pimozide injection on mean LH pulse frequency was greater in early than in late anoestrus, especially in ewes receiving a high plane of nutrition (P < 0·05 and P = 0·07 for H and L ewes, respectively in April and P = 0·07 for H ewes in July). A significant increase of LH pulse amplitude was also detected in early anoestrus in H ewes (P < 0·01). These results provide evidence that endogenous opioid mechanisms are involved in the inhibition ofLH pulsatile release both in early and late anoestrus in ewes with a short seasonal anoestrus. The ability of pimozide to increase LH pulse frequency in early anoestrus could be enhanced by a high plane of nutrition in the breed studied.

The effect of nutritional growth restriction on reproductive development in red deer stags reared in constant photoperiod was investigated and the correlation between reproductive status and circulating concentrations of growth hormone (GH) and insulin-like growth factor-1 (IGF-1) examined.

Stags were reared from birth in constant photoperiod (12 h light: 12 h dark). They were offered a ‘complete diet’ ad libitum until they reached 52-5 kg live weight (LW) and were then maintained at this LWby restricting daily food dry-matter intake (DMI)for either a long (LR, 26 to 51 weeks of age, no. = 5) or short (SR, 21 to 33 weeks, no. = 6) period before being returned to ad libitum feeding.

Relative to SR stags, antlers hardened later in LR stags (72·0 v. 57·5 weeks of age, P < 0·001); sustained elevations in plasma testosterone also began later (55·2 v. 38·5 weeks, P < 0·001) but at lower LW (54·0 v. 60·6 kg, P < 0·05). In both groups, the testosterone rise followed the return to ad libitum food by 4 to 5 weeks. Plasma IGF-1 was closely correlated with DMI (LR, r = 0·84, P < 0·001; SR, r = 0-93, P < 0·001) and with plasma testosterone (LR, r = 0·42, P < 0·001; SR, r = 0·38, P < 0·01). Also the increase in plasma IGF-1 preceded that of testosterone (by 2·8 (s.e. 0·94) weeks) and was associated with a transient plasma GH peak (P < 0·05) and elevated LH concentrations (SR, P < 0·01; LR, P < 0·05).

The timing of reproductive development in growth-restricted male red deer in constant photoperiod may therefore be more sensitive to DMI than LW, and changes in plasma IGF-1 concentrations are consistent with a putative permissive role in relaying this information.

An investigation was conducted to compare, the LH, prolactin and progesterone profiles of ewe lambs with ewes during oestrus and early pregnancy. Peak LH concentration was significantly lower (P < 0·05) and duration of release tended to be shorter during oestrus in ewe lambs. Furthermore, this smaller LH peak tended to be associated with a lower ovulation rate. LH pulse rate, amplitude and concentration were similar in both groups during day 13 of pregnancy. However, LH concentration was higher in ewe lambs compared with ewes on day 28 of pregnancy. Peak prolactin concentration was lower during oestrus in ewe lambs (P < 0·05) but there was no difference between the two groups during early pregnancy. Plasma progesterone concentrations were similar in both groups until day 13 of pregnancy. Thereafter, mean progesterone concentration was lower, from day 14 to 30, in the ewe lambs (P < 0·05). These results indicate that in ewe lambs and ewes maintained under identical conditions plasma hormone profiles differ during oestrus and early pregnancy. Moreover, these differences probably contribute towards the lower reproductive performance of ewe lambs.

An experiment was conducted to examine the effects of body condition at calving and feeding level post partum on the duration of the post-partum anoestrous period in beef cows. Forty-seven cows were used in each of 2 years. Cows were fed differentially before calving to achieve two levels of body condition at calving in November and December and were then fed at one of two levels of feeding post partum in a 2 × 2 factorial experiment. At calving, mean body condition scores were 2·85 (F) and 2·23 (T) (P < 0·001) while live weights were 521 kg and 457 kg (P < 0·001) for F and T cows respectively. After calving metabolizable energy intakes were 111 and 118 MJ/day (H) or 56 and 64 MJ/day (L) in years 1 and 2 respectively. The H cows maintained live weight during the first 61 days of lactation while the L cows lost live weight (0·0 v. -1·1 kg/day, P < 0·001). The number of cows still acyclic on 1 May each year was higher (P < 0·01) on the TL treatment (10/24) compared with the other treatment combinations (FH: 2/22; FL: 5/22; TH: 2/24). The effect of level of feeding on the duration of the anoestrous period was significant in the T cows (89 v. 116 days for TH and TL cows respectively; P < 0·01) but not in the F cows (80 v.89 days for the FH and FL cows respectively; P > 0·05). At 3 weeks post partum LH pulse frequency was higher in F than in T cows and tended to be higher in H than in L cows. At 6 and 9 weeks post partum LH pulse frequency was higher in H than L cows. LH pulse amplitude was higher in F than T cows at 3 weeks post partum and tended to be higher at 6 weeks post partum. It is concluded that both body condition at calving and feeding level after calving influence the duration of the post-partum anoestrous period with the effect of feeding level being particularly pronounced in thin cows.

An experiment was conducted to determine the effect of level and pattern of food intake in post-partum beef cows on the length of the post-partum anoestrous period and associated luteinizing hormone profiles. Forty-six multiparous cows were allocated at calving to one of three feeding regimes: 55 MJ metabolizable energy (ME) per day throughout (LL), 145 M] ME per day throughout (HH), or 55 M] ME per day until 6 weeks post partum and then 145 M] ME per day thereafter (LH). The duration of the post-partum anoestrous period was assessed from plasma progesterone profiles. Luteinizing hormone profiles in the peripheral circulation were determined at weeks 3, 6, 8 and 11 post partum from samples collected every 20 min for 10 h.

Feeding regime had no effect on either the length of the post-partum anoestrous period, being 99, 89 and 91 (s.e.d. 12·2) days for the LL, LH and HH cows respectively, or on any aspect of luteinizing hormone pulsatility. Body condition at calving was negatively correlated with the duration of the post-partum anoestrous period, each unit increase in body condition at calving being associated with a decrease in the post-partum anoestrous period of 86 (s.e. 21·8) days. Luteinizing hormone pulse frequency at 6 weeks post partum was positively correlated with body condition at calving.

This experiment confirms the importance of body condition at calving in influencing the duration of the post-partum anoestrous period and indicates that level and pattern of feeding post partum has little effect when cows calve at a body condition of approximately 2·25.

Pre-pubertal ewe lambs were introduced to rams on 17 August (Tl), 12 September (T2) or 10 October (T3) or isolated from rams until 21 December (Tc) (no. = 13). Blood was taken for progesterone assay three times weekly (no. = 13) from 16 July. Blood was taken for LH assay at 15-min intervals for 8 h (no. = 8) at the time of the introduction of rams and subsequently at the time of further ram introductions. Blood was taken from Tc lambs at each ram introduction. Tl, T2 and T3 lambs ovulated for the first time 16 (s.e. 2·2) days earlier than Tc lambs but only T3 lambs ovulated after and apparently in response to the introduction of the rams. The mean baseline LH levels were higher in Tl, T2 and T3 lambs in comparison with Tc lambs at all sampling periods. The mean pulse frequency of LH secretion was significantly increased by the introduction of rams in T2 and T3 lambs in comparison with Tc lambs. The mean pulse frequency of LH secretion was greater in Tl, T2 and T3 lambs at the October sampling period in comparison to Tc lambs.