Effects of nutritional status and gonadal steroids on expression of appetite-regulatory genes in the hypothalamic arcuate nucleus of sheep.

Sheep exhibit photoperiod-driven seasonal changes in appetite and body weight so that nutritional status increases in long days (LD) and decreases in short days (SD); additionally, they are reproductively active in SD and inactive in LD. We addressed the hypothesis that appetite-regulatory genes in the hypothalamus respond differently to changes in nutritional feedback induced by photoperiod as opposed to food restriction, and that responses would be influenced by gonadal steroid status. Castrated oestradiol-implanted male sheep were kept in SD (8 h light/day) or LD (16 h light/day) for 11 weeks, with ad libitum or restricted food (experiment 1; n=8/group). Rams were kept in SD or LD for 12 weeks with ad libitum or restricted food (experiment 2; n=6/group). Gene expression (by in situ hybridisation) in the hypothalamic arcuate nucleus for leptin receptor (OB-Rb), neuropeptide Y (NPY), pro-opiomelanocortin (POMC) and agouti-related peptide (AGRP) was unaffected by photoperiod treatment, but food restriction increased NPY and AGRP mRNAs, in experiment 1. In experiment 2, mRNAs for POMC and cocaine- and amphetamine-regulated transcript (CART) were up-regulated and AGRP down-regulated in SD, while food restriction increased OB-Rb mRNA, increased NPY and AGRP mRNAs only in LD and decreased POMC mRNA only in SD. Thus, gene expression responded differently to photoperiod and food restriction, and the melanocortin pathway was up-regulated in SD in reproductively activated rams but not in oestradiol-implanted castrates. These data support the hypothesis that hypothalamic appetite-regulatory pathways respond differently to changes in nutritional feedback induced by photoperiod as opposed to food restriction, with gonadal steroid feedback additionally influencing the responses.

The role of cholecystokinin in the expression of seasonal variation in the feed intake and eating pattern of red deer (Cervus elaphus).

The role of cholecystokinin (CCK) in the expression of seasonal variation in feed intake and feeding activity in ruminants was investigated by active immunisation of castrated male red deer (Cervus elaphus). In April, animals of two groups (five animals per group) were immunised against either CCK or vehicle solution only and booster injections were administered at 2-month intervals for the following year. Measurements were conducted for a period of more than a year from July. There were no significant effects of immunisation on mean daily food intake. However, there was a significant interaction (P<.01) between immunisation and month (season), with respect to rate of feed ingestion during meals, with animals immunised against CCK exhibiting higher mean rates of ingestion during October to May but lower mean rates during June to September. It is concluded that systemic CCK has a role in the expression of seasonal variation in the rate of feed ingestion during meals in ruminants and that this, in turn, may affect the pattern of seasonal change in daily feed intake. Since there was no evidence of differences with treatment in profiles of insulin, growth hormone (GH), thyroxine (T(4)), triiodothyronine (T(3)), insulin-like growth factor-1 (IGF-1) or prolactin, it is unlikely that this effect is expressed through changes in the profiles of these hormones.

Effect of long-term feed restriction on seasonal endocrine changes in soay sheep.

Groups of 15 adult, castrated, male Soay sheep were housed under natural daylength conditions at 57 degrees N and fed a complete diet ad libitum (AL) or at a restricted rate (R) of 35 g dry matter (DM)/kg(0.75) initial liveweight per day. The diet was based on barley and dried grass pellets and contained an estimated 11.6 MJ of metabolisable energy, 83% DM and 140 g crude protein/kg DM. In the AL animals, higher levels of feed intake during the periods of long daylength were associated with shorter inter-meal intervals (p<0. 001), a greater meal frequency (p<0.001), and a greater proportion of time spent eating (p<0.001) together with a greater rate of feed ingestion (p<0.001) and an increased meal size (p<0.001). Mean plasma concentrations of insulin, prolactin, insulin-like growth factor 1 (IGF-1), triiodothyronine (T(3)), and thyroxine (T(4)) were higher (p<0.001) in the spring or summer than in the autumn. Mean plasma GH concentrations did not differ with month. Compared with R animals, AL animals had higher mean plasma concentrations of insulin (p<0.001), prolactin (p<0.01), T(3) (p<0.01), and T(4) (p<0.01). Plasma GH and IGF-1 concentrations did not differ significantly with treatment. There was a greater increase in plasma insulin concentrations following feeding in R than AL animals (p<0.001) owing to higher pre-feeding concentrations in AL animals and the ingestion of larger amounts of feed by R than AL animals in the period after fresh feed was introduced. There were significant differences between months in this response, in R animals (p<0.01). Mean CSF insulin concentrations were significantly higher in AL than R animals (p<0.05) but were not affected by month. Neither was there a difference between pre-feeding concentrations and concentrations at approximately 12 h after feeding. It is concluded that the differences in the response of plasma insulin concentrations to feeding at different times of year, which were detected in R animals, were attributable, primarily, to differences in the vagally-induced insulin response to feeding and that these differences may provide important feedback signals to the appetite centre.

Effects of pulsed or continuous infusion of cortisol on immune function in sheep.

It was postulated that frequent pulses of cortisol such as might be induced by a repeated or chronic stressor, could induce immune suppression and that the effect would be greater than in animals subjected to less frequent increases. Four groups of nine adult Scottish Blackface ewes were infused for 14 d with saline or hydrocortisone hemisuccinate (cortisol) delivered continuously or in pulses. Plasma concentrations of cortisol were significantly elevated (to between approximately 100 and 1000 nmol/liter; P < 0.001) for about 30 or 75 min after infusion of pulses of hydrocortisone hemisuccinate at intervals of 1 hr (P1) or 6 hr (P6), respectively. In animals continuously infused (CI), they were consistently elevated (P < 0.001), compared with concentrations in control animals infused with saline only (S), to approximately 1000 nmol/liter or more. Antibody production in response to ovalbumin injection was not affected by any of the infusion regimes. At Days 10, 24, and 31 after injection of ovalbumin and initiation of the infusion, rates of multiplication of unstimulated lymphocytes, in vitro, were greater (P < 0.05) in P6 animals than in saline-infused, control animals and this resulted in a reduction in the stimulated lymphocyte response. As a consequence of the increased basal lymphocyte activity, after Day 0, the corrected, stimulated lymphocyte response of P6 animals was consistently below that of controls (P < 0.05 at Day 24). Both mean basal and stimulated lymphocyte activities in CI and P1 animals were similar to those of controls. The gamma interferon (IFN-gamma) response was generally small and not affected by treatment. It is concluded that large, relatively infrequent increases in circulating cortisol concentrations can modify the cell mediated immune response such that the response to a specific antigen challenge is compromised but smaller, more frequent pulses had no effect. Elevated cortisol concentrations per se did not have a significant inhibitory effect on the immune system.

Seasonality of meal patterns and hormonal correlates in red deer.

Two groups of six adult, castrated, male red deer were housed under natural daylength conditions and at ambient temperature at 57 degrees N and fed ad lib. (AL) or at a fixed rate of 50 g/kg0.75 initial liveweight per day throughout the study (restricted, R). Mean daily intakes of AL animals were higher during periods of long daylength than during short daylength (p < 0.001). The higher rates of food intake during periods of long daylength were a function of greater meal durations (p < 0.001), shorter inter-meal intervals (p < 0.001) and higher (p < 0.001) mean rates of ingestion (g/min). In both groups mean plasma concentrations of prolactin, T3, T4, and insulin-like growth factor 1 (IGF-1) were higher (p < 0.001) during long daylength than short daylength although changes in thyroid hormone profiles were much less marked in AL animals. Insulin and growth hormone (GH) profiles exhibited no consistent seasonal trend. Mean plasma concentrations of T3 were higher in AL than in R animals. Mean plasma IGF-1 concentrations during long days were consistently greater in the AL than R animals. It is concluded that the effects of seasonal changes in daylength on appetite and food intake are expressed through changes in both the duration of daylight periods per se and in underlying seasonal changes in physiology and associated meal patterns and eating rates. It is concluded that the roles of T3, IGF-1, and prolactin in the expression of seasonal changes in appetite should be investigated further and, particularly, their effects on other hormone profiles and liver and gut function.

Effect of undernutrition of ewes from the time of mating on fetal ovarian development in mid gestation.

Ewes were fed either 150% (High, H) or 50% (Low, L) of their energy requirements for maintenance of liveweight during early gestation. Effects of maternal nutrition on fetal ovarian size, histological structure and steroidogenic capacity were studied at Day 47 and on ovarian size and structure at Day 62 of gestation. At Day 47 of gestation, there were significantly higher concentrations of oogonia in the ovaries of L fetuses than H fetuses (105.9 v. 76.9 germ cells mm(-2); s.e. 4.94; P < 0.001). The capacity of the ovaries to secrete oestradiol (pg/ovary/24 h) at Day 47 was not affected by treatment when they were incubated either with (H, 773; L, 740; s.e. 179; not significant, n.s.) or without (H, 260; L, 290; s.e. 92.7; n.s.) ovine luteinizing hormone (oLH). At Day 62 of gestation, the process of germ cell degeneration was less advanced in L than H fetal ovaries, as indicated by higher oocyte concentrations in the former (68.4 v. 48.6 germ cells mm(-2); s.e. 3.85; P < 0 01). There was a greater percentage of meiotic cells in L ovaries (76.5 v. 18.6; s.e. 5.82; P < 0.001). It is concluded that undernutrition of the ewe from the time of mating significantly retards ovarian development in fetal ovaries.

Effects of bovine somatotrophin (bST) on ovarian function in post-partum beef cows.

The effects of bovine somatotrophin (bST) on ovarian follicle development and function and associated gonadotrophin profiles during the first nine weeks post partum were investigated in beef cows. Thirty-two cows (Shorthorn x Galloway) in moderately low body condition (BC) at calving were fed to maintain BC thereafter. At Weeks 2, 4, 6 and 8 post partum, animals were injected with 320 mg bovine somatotrophin (bST) (T, treated; n = 17) or with the carrier oil only (C, control; n = 15). Ovulation occurred in 4 of 17 T cows and 0 of 15 C cows (P = 0.10) by nine week post partum. Treatment with bST did not affect the numbers of small (3-8 mm in diameter) or large (> 8 mm in diameter) follicles or the granulosa cell populations but enhanced the oestradiol (P < 0.05) and insulin-like growth factor-I (IGF-I) content (P < 0.01) of large follicles by nine weeks post partum. It did not significantly affect the testosterone concentrations of large follicles. Circulating concentrations of growth hormone (GH) and IGF-I were higher in T cows than in C cows (P < 0.001) but were unrelated to gonadotrophin profiles or gonadotrophin receptor concentrations in the follicles. At Week 8, plasma insulin concentrations were higher in T cows than in C cows both before (P < 0.05) and after (P < 0.05) glucose injection. It is concluded that GH may play an important role in mediating the effects of nutritional state on ovarian function during the post-partum period, possibly through alteration of intrafollicular IGF-I concentrations.

FSH-stimulated follicle development in ewes in high and low body condition and chronically treated with gonadotrophin-releasing hormone agonist.

Ovarian follicle development in response to FSH infusion was investigated in Scottish Blackface ewes with high and low body condition scores in which endogenous gonadotrophin secretion and follicle development to > or = 2.5 mm diameter was suppressed using subcutaneous implants containing a GnRH agonist. In two experiments conducted during the normal breeding season, groups of 20 (Expt 1) and 15 (Expt 2) ewes were fed to achieve body condition scores > or = 2.75 (high; H) or < or = 1.75 (low; L). In both experiments GnRH agonist implants were inserted four weeks before FSH was infused for 72 h at 7 micrograms h-1 to group H animals or at 5 micrograms h-1 to group L animals; the infusion rates were designed to ensure similar circulating FSH concentrations in animals of both groups. In Expt 2, additional subcutaneous implants containing oestradiol were inserted 21 days after insertion of GnRH agonist implants and 7 days before the FSH infusion began. In both experiments, FSH infusion was associated with an increase in circulatory concentrations of LH (P < 0.01) and FSH (P < 0.001), but there was no difference with body condition in mean circulating gonadotrophin concentrations, the numbers of ovarian follicles > or = 2.5 mm diameter, the proportion of these follicles that were oestrogenic or the mean rate of oestradiol secretion in vitro. It is concluded that differences in body condition of ewes do not affect the responsiveness of the ovary to FSH, in the presence or absence of oestradiol, as measured by the number, size and steroidogenic capacity of ovarian follicles present following FSH infusion.

FSH and LH receptor concentrations in large ovarian follicles of beef cows in high and low levels of body condition at nine weeks post partum.

An experiment was conducted to determine whether or not the effect of body condition at parturition, and thereafter, on the duration of the post-partum anoestrous period was mediated through differences in gonadotrophin receptor concentrations in the ovarian follicles. Two groups of cows were fed to achieve body condition scores of > or = 2.75 (H; n = 14) or < or = 2.00 (L; n = 15) at parturition (this had led to differences in the duration of post-partum anoestrus in previous experiments) and then fed to maintain liveweight and condition until ovariectomy at nine weeks post partum. There was no effect of body condition on the mean (+/- s.e.) number of large (> or = 8 mm diameter) follicles per cow (3.0 +/- 0.27) but a greater proportion of the follicles of H cows, compared with L cows, was oestrogenic (secreting > 250 pg oestradiol h-1 in culture) (0.75 v. 0.5; chi 2 = 3.40; P = 0.06). Mean concentrations (pg hormone bound per mg protein) of follicle stimulating hormone (FSH) receptors in both granulosa (H, 157.3; L, 136.6; standard error of difference (s.e.d.) = 27.2) and thecal tissue (H, 4.14; L, 4.35; s.e.d. = 1.66) were similar for H and L cows. There was no difference associated with treatment in luteinizing hormone (LH) receptor concentrations in either granulosa (H, 40.3; L, 37.2; s.e.d. = 6.67) or thecal tissue (H, 48.9; L, 48.0; s.e.d. = 6.03). The concentration of FSH receptors in granulosa tissue was lower in non-oestrogenic follicles than in oestrogenic follicles (80.0 v. 188.1; s.e.d. = 40.4; P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)