T.E. Kiser

Growth Hormone-Releasing Hormone and Somatostatin Neurons within the Porcine and Bovine Hypothalamus

Hypothalamic growth hormone-releasing hormone (GHRH) and somatotropin release-inhibiting factor or somatostatin (SS) immunoreactive (ir) neurons were localized in pigs (n = 8) and cattle (n = 7) to identify neuroanatomical sites involved in the regulation of growth hormone secretion. Coronal and sagittal frozen sections (30-60 microns) of Zambonis fixed hypothalamic tissue, without prior colchicine treatment were incubated with GHRH or SS primary antisera for 48 h, then visualized by peroxidase-diaminobenzidine immunocytochemistry. Fusiform, bipolar SS-ir perikarya were located about the third ventricle in the periventricular nucleus, extending from rostral aspects of preoptic periventricular nucleus to a level approximate with caudal regions of the paraventricular nucleus. Rounded or fusiform, bipolar GHRH-ir perikarya were mostly located in ventrolateral portions of the arcuate nucleus in pigs and cattle, and within ventral aspects of the ventromedial nucleus in pigs but rarely in cattle. In both pigs and cattle, SS-ir and GHRH-ir fibers projected ventrally into the median eminence with dense and overlapping innervation of the external layer, especially dense in lateral regions. In pigs, but not as distinguishable in cattle, SS-ir fibers also densely innervated the ventromedial and arcuate hypothalamic nuclei. Double immunostained sections revealed close apposition of SS-ir fibers and varicosities with GHRH-ir perikarya in arcuate and ventromedial nuclei, and apposition of SS-ir and GHRH-ir varicosities in the median eminence.

The effect of calf removal on estrous response and pregnancy rate of beef cows after SYNCRO-MATE-B treatment☆☆☆★

Abstract The effect of calf removal on estrous response and pregnancy rate of cows given SYNCRO-MATE-B (SMB) was determined. Two hundred forty three lactating beef cows treated with SMB were allotted to three treatments: treatment 1, no calf removal; treatment 2, 24 hr calf removal and treatment 3, 48 hr calf removal. Calf removal time began with removal of the SMB implant. All cows were artificially inseminated 48 to 54 hr after implant removal. Body condition of cows in each heard was visually appraised as adequate or marginal. At each location, technicians and sires were stratified across all treatments. Estrous response of cows (n=143) was different (P In summary, estrous response and pregnancy rate of lactating beef cows treated with SMB were increased by 48 hr calf removal, compared to no calf removal or 24 hr calf removal, when calf removal time began with removal of the SMB implant.

Effect of nutrition on the LH response to calf removal and GnRH

Ten primiparous crossbred cows were assigned to two dietary groups at calving. One group received 120% and the other group received 80% of the National Research Council (NRC) recommended allowance of dietary energy for primiparous cows. At 60 days postpartum, calves were removed from their dams. Blood samples were collected from the cows at 15-min intervals for 8 hr beginning at the time of calf removal and again 24 hr, 48 hr and 72 hr after calf removal. At 72 hr after calf removal, all cows were given 200 ug GnRH intravenously. At calf removal, serum LH concentrations were higher (P<0.01) for cows on 120% (0.9 +/- 0.03 ng/ml) compared to cows on 80% (0.5 +/- 0.03 ng/ml) of recommendations. Serum LH concentrations increased (1.6 +/- 0.1 ng/ml, P<0.01) by 24 hr in cows on the highenergy diet. In contrast, a similar increase was not observed in cows on the low-energy diet until 48 hr after calf removal (1.4 +/- 0.2 ng/ml, P<0.01). These contrasting patterns in serum LH concentrations resulted in a diet by time interaction (P<0.01). Serum LH concentrations increased in both dietary energy groups following GnRH injection, but the response was greater (P<0.01) in cows on the low-energy diet compared to the cows fed the high-energy diet. These results indicate that inadequate dietary energy delays the LH response to calf removal and increases the LH response to exogenous GnRH.

Naloxone infusion increases pulsatile luteinizing hormone release in postpartum beef cows

The limiting factor in the return to cyclicity in the postpartum cow appears to be the lack of pulsatile luteinizing hormone (LH) secretion. To test the role of endogenous opioids in regulating pulsatile LH release, naloxone, an opioid antagonist, was infused into postpartum cows. Eight cows (39.3 ± 2.1 d postpartum) received either a constant infusion of saline or 50 mg/hr of naloxone dissolved in saline for eight hr. Blood samples were taken at 15 min intervals for determination of serum LH concentrations and to determine frequency and amplitude of the LH pulses. Frequency of LH pulses was greater (P .05). Mean serum LH concentrations were greater (P<.01) in the naloxone infused group (2.7 ± .3 ng/ml) compared to the saline infused group (1.9 ± .4 ng/ml). These data indicate that endogenous opioids inhibit pulsatile LH secretion in the anestrous postpartum beef cow and that naloxone infusion increases mean serum LH concentrations by increasing the frequency of LH pulses.

Delayed puberty in gilts in total confinement.

Two hundred fourteen crossbred gilts, born in January through March and June through July of two different years, were raised in total confinement until 100 to 120 days of age and then moved to an outside dirt lot (non-confined) or to a single pen in a confinement, finishing building (confined). Beginning at 150 days of age, estrus was checked daily with a boar to determine percentage of gilts that attained puberty and age at first estrus, and weekly blood samples were collected and analyzed for progesterone by radioimmunoassay to determine age at first ovulation. In the Jan.-Mar. born gilts, 75.4% of the non-confined gilts and 37.4% of the confined gilts attained puberty by 270 days of age (P<.001). Although differences were not significant in the gilts born in June-July, more non-confined gilts (62.6%) than confined gilts (50.9%) attained puberty. Of the 121 gilts that ovulated, only 1 non-confined and 3 confined gilts did not exhibit estrus. Average age at first estrus or at first ovulation were similar for confined and non-confined gilts. Adrenal gland weights at 250 to 270 days of age were similar also for confined and non-confined gilts. Based on the results of this study, we conclude that total confinement housing can reduce, by as much as 50%, the proportion of gilts that attain puberty by 8 to 9 months of age and that time of year may influence the extent of delayed puberty.

Immunocytochemical localization of the catecholamine-synthesizing enzymes, tyrosine hydroxylase and dopamine-β-hydroxylase, in the hypothalamus of cattle ☆

Immunocytochemical staining for the presence of catecholamine synthesizing enzymes, tyrosine hydroxylase and dopamine beta-hydroxylase, was used to characterize the regional distribution of catecholaminergic neurons in the hypothalamus and adjacent areas of domestic cattle, Bos taurus. In steers, heifers and cows, tyrosine hydroxylase-immunoreactive perikarya was located throughout periventricular regions of the third cerebral ventricle, in both anterior and retrochiasmatic divisions of the supraoptic nucleus, suprachiasmatic nucleus, and ventral and dorsolateral regions of the paraventricular nucleus, dorsal hypothalamus, ventrolateral aspects of the arcuate nucleus, along the ventral hypothalamic surface between the median eminence and optic tract, and in the posterior hypothalamus. Immunostained perikarya ranged from small (10-20 microns, parvicellular) to large (30-50 microns, magnocellular) and were of multiple shapes: round, triangular, fusiform or multipolar, often with 2-5 processes of branched arborization. There were no dopamine-beta-hydroxylase immunoreactive perikarya observed within the hypothalamus and adjacent structures. However, both tyrosine hydroxylase and dopamine-beta-hydroxylase immunoreactive fibers and punctate varicosities were observed throughout regions of tyrosine hydroxylase immunoreactivity perikarya. Generally, the location and pattern of hypothalamic tyrosine hydroxylase immunoreactivity and dopamine-beta-hydroxylase immunoreactive were similar to those reported for most other large brain mammalian species, however, there were several differences with commonly used small laboratory animals. These included intense tyrosine hydroxylase immunoreactivity of perikarya within the retrochiasmatic division of the supraoptic nucleus (ventral A15 region), the absence of tyrosine hydroxylase immunoreactive perikarya below the anterior commissure or within the bed nucleus of stria terminalis (absence of the dorsal A15 region), an abundance of tyrosine hydroxylase immunoreactive perikarya within the ependymal layer of the median eminence, heavy innervation of the arcuate nucleus with dopamine-beta-hydroxylase immunoreactive fibers and varicosities, and the paucity of dopamine-beta-hydroxylase immunoreactive throughout the median eminence.

Effect of calf removal on serum luteinizing hormone and cortisol concentrations in postpartum beef cows

Serum luteinizing hormone (LH) and cortisol concentrations were measured in ten fall calving, Angus cows averaging 38 +/- 8 days postpartum. Calves from five cows were weaned at the beginning of the study. Blood samples were collected at 20 min. intervals for 48 h after weaning and for 8 h on day 4 and day 6 postweaning. Mean serum LH concentrations increased (P<0.01) in weaned cows (W) from 0.55 +/- 0.01 ng/ml at time of calf removal to 1.3 +/- 0.04 ng/ml 48 h afterwards. Comparable LH concentrations for suckled cows (S) were 0.65 +/- 0.08 ng/ml and 0.62 +/- 0.03 ng/ml respectively. Average serum LH concentrations at 48 h after weaning were greater (P<0.01) for W cows than S cows and a treatment by time interaction occurred (P<0.01) with serum LH concentrations increasing (P<0.01) from time of calf removal to 48 h after calf removal in W cows. Frequency of LH peaks increased (P<0.01) in W cows and by 48 h after weaning was greater (P<0.01) in W cows than in S cows. Magnitude of LH peaks did not differ between the two groups. Serum cortisol concentrations were not different between W and S cows except for a transient elevation (P<0.01) in W cows from 7.6 +/- 0.9 ng/ml to 11.9 +/- 1.0 ng/ml 9 to 12 h after calf removal. Since serum LH concentrations were increased in W cows but not in S cows at 48 h and serum cortisol concentrations increased transiently in W cows we suggest that circulating cortisol levels may not be a physiological inhibitor of LH secretion in the suckled postpartum beef cow.

Suckling induced cortisol secretion in young beef cows.

The profiles of plasma cortisol concentration in response to suckling were determined in 10 young, postpartum beef cows between days 25 and 85 postpartum. Two trials, comprised of five cows each, were conducted in the fall (I) and spring (II), respectively. In both trials, plasma cortisol rose within 10 minutes after suckling began and was significantly higher than pre-suckling concentrations (P<.01). Over the next 30 minutes in trial I and 40 minutes in trial II, the cortisol level progressively fell back to the pre-suckling levels. This profile was qualitatively similar among the days postpartum on which the cows were bled. Neither the profile nor the peak concentration after suckling changed significantly (P>.10) as days postpartum lapsed. Finally, there was a significant difference (P<.01) in mean plasma cortisol between the cows in trial I compared to the cows in trial II.

Serum progesterone concentrations in pregnant and nonpregnant heifers and after gonadotropin releasing hormone in luteal phase heifers

Abstract Serum progesterone (P4) concentrations were quantitated in 18 Holstein heifers from days 5 to 16 after estrus in an effort to ascertain the effects of pregnancy on circulating levels of this hormone. The P4 concentration rose faster between days 5 and 10 in the pregnant heifers compared to P4 levels in both the non-pregnant heifers and the killed sperm inseminated group. It was found that serum P4 levels were significantly (P The administration of 250μg gonadotropin releasing hormone (GnRH) to 10 Holstein heifers on day 7 after estrus resulted in a significant (P

Prostaglandin F concentrations in utero-ovarian vein plasma of prepuberal and mature gilts.

Abstract Prostaglandin F (PGF) and progestins in utero-ovarian vein (UOV) plasma during the late luteal phase of the estrous cycle in unbred mature gilts and following induced ovulation in unbred prepuberal gilts were determined. Prepuberal gilts (120 to 130 days of age) were induced to ovulate with Pregnant Mare Serum Gonadotropin and Human Chorionic Gonadotropin (HCG). The day following HCG was designated as Day 0. Mature gilts which had displayed two or more estrous cycles of 18 to 22 days (onset of estrus = Day 0) were used. Polyvinyl catheters were inserted into the UOV of all gilts and blood was collected at 15 min intervals from 0800 to 1045 hr on Days 10 through 20 or Days 12 through 18. Plasma PGF concentrations in the mature gilts were elevated on Days 13, 14, 15, 16 and 17, whereas, plasma PGF concentrations in the prepuberal gilts were elevated only on Days 15, 16 and 17 resulting in a reproductive age (mature vs prepuberal) by day interaction (P