George B. Rampacek

Developmental Changes in the Long Form Leptin Receptor and Related Neuropeptide Gene Expression in the Pig Brain

Abstract The hypothalamus is the key site of central regulation of energy homeostasis, appetite, and reproduction. The long form leptin receptor (Ob-Rl) is localized within the hypothalamus along with several neuropeptides that are involved in regulation of the neuroendocrine axis. In the present study, developmental changes in gene expression of the Ob-Rl, preproorexin, proopiomelanocortin (POMC), corticotropin releasing factor (CRF), somatostatin, and GnRH in the hypothalamus was studied. Expression of Ob-Rl and neuropeptide mRNA was examined by semiquantitative reverse transcription-polymerase chain reaction in hypothalami collected from 106-day-old fetus (n = 3) and 7-day-old (n = 3), 3.5-mo-old (n = 3), and 6-mo-old (n = 2) gilts. In addition, leptin mRNA expression in the first three ages was examined in back fat. Leptin mRNA expression increased (P < 0.05) by 7 days postnatal, but Ob-Rl mRNA expression increased (P < 0.01) by 3.5 mo. Expression of preproorexin (P < 0.05), somatostatin, and GnRH (P < 0.01) mRNA peaked by 3.5 mo of age while POMC mRNA expression increased markedly (P < 0.01) by 6 mo of age. The CRF mRNA expression did not change across ages. These findings suggest a possible relationship among Ob-Rl and a number of hypothalamic and peripheral peptides in the development of the neuroendocrine axis. These peptides may serve as messengers that link mechanisms that regulate reproduction and energy balance.

Effect of suckling on cortisol, progesterone and luteinizing hormone in postpartum beef cows

Five primiparous, 3-year-old Hereford cows suckled ad libitum , were cannulated via the jugular vein and stanchioned for 2-day sampling periods, every 14 days starting 14 days after the mean calving date. On the second day of each period, calves were removed to a pen away from the cows, for 9 hours. Blood was sampled 5 min before calves were returned to their dams, as soon as possible after initiation of suckling (IOS), and at 15-min intervals for 45 min, thereafter. Cortisol, progesterone and luteinizing hormone (LH) concentrations in the serum were quantitated by radioimmunoassay. Mean serum cortisol concentrations were 7.3 +/- .7, 9.4 +/- .7, 12.1 +/- .9, 7.5 +/- .5 and 5.7 +/- .4 ng/ml (mean +/- S.E.) at -5, 0, 15, 30 and 45 min after IOS, respectively, for all cows across all periods. Cortisol concentrations, during and after suckling, tended (P<.06) to differ among sampling periods, during the postpartum interval. Serum progesterone concentrations were .28 +/- .02, .28 +/- .02, .32 +/- .05 and .24 +/- .03 ng/ml at 0, 15, 30 and 45 min after IOS, respectively, for all cows across all period, indicating that suckling had no effect on serum progesterone, and were similar at all sampling periods during the postpartum interval. Serum LH concentrations were .81 +/- .07, .77 +/- .06, .71 +/- .04, and .72 +/- .04 ng/ml at 0, 15, 30 and 45 min after IOS, respectively. During the postpartum interval, serum LH concentrations were greater (P<.01) at 71 and 85 days postpartum than at any other time.

Synchronization of estrus and ovulation in gilts with estradiol and prostaglandin F2α

Abstract Twenty-two crossbred gilts at various stages of the estrous cycle were injected IM with 5 mg of estradiol benzoate (EB) daily for 20 days. Five days after the last EB injection, 11 gilts were injected IM with 10 mg of prostaglandin F 2α (PGF 2α ) free acid and 11 gilts were injected IM with saline (control). Gilts were bred at first estrus, and ova were recovered 4 days later from PGF 2α - treated gilts. The controls failed to display estrus throughout the experimental period, and at necropsy, corpora lutea (CL) were present in 10 control gilts. The other control gilt had preovulatory follicles. Nine of the 11 PGF 2α - treated gilts displayed estrus 6.1 ± .4 days after treatment. However, one of these nine gilts failed to ovulate and only preovulatory follicles were present on the ovaries. The two PGF 2α - treated gilts that remained anestrual throughout the experiment had only preovulatory follicles present on the ovaries. Plasma progesterone levels of control gilts did not change throughout the experimental period while progesterone levels dropped sharply from approximately 14 ng/ml to 1 ng/ml within 24 hr after PGF 2α injection and remained low throughout the sampling period indicating that progesterone production of the CL maintained by the EB was suddenly terminated by PGF 2α . The number of ova recovered was 87% of the number of CL present and 77% of the ova recovered were fertilized. Ovarian cysts were absent and pretreatment ovulation rate of the controls and post-treatment ovulation rate of the PGF 2α gilts were similar. The results from this experiment suggest that estrogen pretreatment of randomly cycling gilts for 20 days followed by PGF 2α is an effective estrus and ovulation control regime.

Luteinizing hormone and prolactin secretion in hypophysial-stalk-transected pigs given estradiol and pulsatile gonadotropin-releasing hormone

To determine the sites of action whereby estradiol (E2) induces the preovulatory luteinizing hormone (LH) surge in pigs, hypophysial-stalk-transected (HST) ovariectomized (OVX) gilts (mean +/- SE: 118 +/- 12 kg) received intramuscular injections of E2 benzoate (10 micrograms E2B/kg body weight) on day 0, plus intravenous pulses of 1 microgram gonadotropin-releasing hormone (GnRH) every 45 min either from days -5 to 4, days -5 to 0 or days -5 to 0 then days 2 to 4. A fourth and fifth group received steroid vehicle on day 0 and pulses of GnRH on either days -5 to 4 or days -5 to 0 then days 2 to 4. In stalk-intact OVX gilts, E2B inhibited LH release for 48 h, then induced an LH surge that peaked 60-84 h after steroid treatment. In HST OVX gilts, serum LH increased from undetectable concentrations (less than 0.17 ng/ml) to 0.42 +/- 0.03 ng/ml after 5 days of pulsatile GnRH replacement. In the presence of pulsatile GnRH stimulation, serum LH concentrations were inhibited for 12 h after E2B but then returned to values that were similar to those for HST gilts given GnRH in the absence of steroid treatment. Discontinuing GnRH pulses at the time of E2B injection caused serum LH concentrations to drop to 0.18 ng/ml or less for the remaining 96 h. When GnRH pulses were interrupted for 48 h, beginning immediately after injecting E2B or vehicle to mimic the secretory hiatus of LH and presumably GnRH observed in stalk-intact pigs prior to the surge, resumption of GnRH pulses caused serum LH concentrations to increase progressively over 3 h in E2B-treated gilts but to peak abruptly in controls given vehicle.(ABSTRACT TRUNCATED AT 250 WORDS)

Luteinizing Hormone Secretion in Hypophysial Stalk-Transected Pigs Given Progesterone and Pulsatile Gonadotropin-Releasing Hormone

Abstract This study was conducted to determine whether progesterone inhibits luteinizing hormone (LH) secretion in female pigs by a direct action on the pituitary gland. Eight ovariectomized, hypophysial stalk-transected gilts were given 1-μg pulses of gonadotropin-releasing hormone iv every 45 min from Day 0 to 12. On Days 5–12, each of four gilts received either progesterone or oil vehicle im at 12-hr intervals. Serum progesterone concentrations in steroid-treated gilts reached 70 ± 6.8 ng/ml (mean ± SE) by Day 8 and remained elevated thereafter, whereas serum progesterone concentrations in oil-treated controls were less than 1 ng/ml for the entire study. Daily serum LH concentrations were not different between gilts treated with progesterone or oil. The 1-μg pulses of gonadotropin-releasing hormone reliably evoked pulses of LH in both treatment groups. The LH pulse frequency and amplitude, assessed from samples collected every 15 min for 6 hr on Day 12, were similar for progesterone- and oil-treated gilts. These results provide evidence that progesterone does not act at the pituitary gland to alter LH secretion in pigs.