Estimation of inbreeding and effective population size of full-blood Wagyu cattle registered with the American Wagyu Cattle Association.

The objective of this research was to examine the population structure of full-blood (100%) Wagyu cattle registered in the United States with the American Wagyu Association, with the aim of estimating and comparing the levels of inbreeding from both pedigree and genotypic data. A total of 4132 full-blood Wagyu cattle pedigrees were assessed and used to compute the inbreeding coefficients (FIT and FST ) and the effective population size (Ne ) from pedigree data for the period 1994 to 2011. In addition to pedigree analysis, 47 full-blood Wagyu cattle representing eight prominent sire lines in the American Wagyu cattle population were genotyped using the Illumina BovineSNP50 BeadChip. Genotypic data were then used to estimate genomic inbreeding coefficients (FROH ) by calculating runs of homozygosity. The mean inbreeding coefficient based on the pedigree data was estimated at 4.80%. The effective population size averaged 17 between the years 1994 and 2011 with an increase of 42.9 in 2000 and a drop of 1.8 in 2011. Examination of the runs of homozygosity revealed that the 47 Wagyu cattle from the eight prominent sire lines had a mean genomic inbreeding coefficient (FROH ) estimated at 9.08% compared to a mean inbreeding coefficient based on pedigree data of 4.8%. These data suggest that the mean genotype inbreeding coefficient of full-blood Wagyu cattle exceeds the inbreeding coefficient identified by pedigree. Inbreeding has increased slowly at a rate of 0.03% per year over the past 17 years. Wagyu breeders should continue to utilize many sires from divergent lines and consider outcrossing to other breeds to enhance genetic diversity and minimize the adverse effects of inbreeding in Wagyu.

LHRH fusion protein immunization alters testicular development, ultrasonographic and histological appearance of ram testis.

This study was designed to evaluate the effectiveness of a luteinizing hormone releasing hormone (LHRH) fusion protein immunization on reproductive traits in ram lambs including the changes in histology and ultrasonographic appearance of testis. Thirty native ram lambs at 19 weeks of age were divided into control (C, n = 10), immunization (I, n = 10) and castration (E, n = 10) groups. Animals in immunization group were immunized against LHRH using ovalbumin-LHRH-7 (OL) protein generated by recombinant DNA technology as a primary and a booster injection at 19 and 23 weeks of age respectively. Animals were bled via jugular venepuncture at 2-week intervals to determine LHRH antibody and testosterone concentrations. Bi-weekly ultrasonographic examination of the testes was performed to determine the changes in ultrasonographic appearance as the age increased. Biopsied testicular tissues taken at 19, 29 and 41 weeks age were also evaluated. At 41 weeks of age, animals were slaughtered. Semen and epididymis were evaluated for the presence of sperm cells. Testicular development and sperm production were suppressed in the immunized animals. Semineferous tubule diameters decreased, basal membrane of the tubule was thickened and hyalinized in immunized ram lambs. While testes of control animals gained their normal ultrasonographic appearance as the age increased, immunized animals had uniform hypoechogenic testicular structure as observed at 19 weeks of age until slaughter. Simultaneous histological and ultrasonographic evaluations indicated that the changes in testicular histology could partly be monitored via ultrasonographic imaging. Nevertheless, it is difficult to claim that ultrasonographic image reflects the exact changes in such instances. In conclusion, these results indicate that recombinant OL fusion protein is effective in immunocastration in ram lambs and has a potential to be used as an alternative to physical castration. Further research studies should be conducted to help assess reproductive status of testes from ultrasound images.

Changes in testicular development, ultrasonographic and histological appearance of the testis in buck kids immunized against LHRH using recombinant LHRH fusion protein.

This study was designed to evaluate the effectiveness of recombinant Ovalbumin-LHRL (OL) immunization on changes in testicular size, histological appearance and testosterone production in buck kids. Thirty native buck kids at 18 weeks of age were divided into three groups, control (n = 10), immunization (n = 10) and castration (n = 10) groups. Immunized animals received OL protein generated by recombinant DNA technology. Ultrasonographic and histological examinations of the testes were performed. Animals were slaughtered at 44 weeks of age. Semen and epididymides were evaluated for the presence of sperm cells. Immunized animals generated anti-LHRH antibodies. Testosterone production, testicular and accessory glands development and sperm production were suppressed in the immunized animals (p < 0.01). Semineferous tubule diameters decreased (p < 0.01), basal membrane of the tubule was thickened and hyalinized in immunized kids. Immunization affected ultrasonographic appearance of the testes drastically. While testes of control animals gained their normal ultrasonographic appearance as the age increased, immunized animals had uniform hypoechogenic testicular structure as observed at 18 weeks of age until slaughter. Simultaneous histological and ultrasonographic evaluations indicated that the changes in testicular histology could partly be monitored via ultrasonographic imaging; nevertheless, it is difficult to claim that ultrasonographic image reflects the exact changes in such instances. In conclusion, these results indicate that recombinant OL fusion protein is effective in immunocastration in buck kids and has a potential to be used as an alternative to physical castration. Further researches should be conducted to help assessing reproductive status of testes from ultrasound images.

Dissociated changes of pituitary luteinizing hormone-releasing hormone (LHRH) receptors and responsiveness to the neurohormone induced by 17 beta-estradiol and LHRH in vivo in the rat.

A single injection of 17 beta-estradiol into castrated male or female rats results in an initial decrease in plasma concentrations of LH and pituitary responsiveness to LHRH, followed by a rapid return to normal or slightly elevated values. Under such experimental conditions, no acute change of binding of [125I-labeled D-Ser(TBU)6]LHRH ethylamide to anterior pituitary homogenate could be observed. Moreover, the self-priming effect of LHRH, as illustrated by a 10-fold increase in the LH response to a second injection of LHRH in the afternoon of proestrus, is accompanied by a 40% loss of pituitary LHRH receptors. During the estrous cycle, a 100% increase in pituitary LHRH receptors is already found on diestrus II, while the maximal LH responsiveness to LHRH occurs later, namely on the afternoon of proestrus. The present findings of a dissociation between changes in LHRH receptor levels and LH responsiveness to the neurohormone suggest that postreceptor events play a predominant role in the control of gonadotropin secretion by sex steroids and LHRH itself. Moreover, LHRH can cause an acute down-regulation of its own receptor in the anterior pituitary gland.

Ewe luteal function influenced by pulsatile administration of synthetic LHRH/FSHRH.

The influence of repetitive administration of synthetic LHRH/FSHRH or saline (S) on 17beta-estradiol (E2)-induced precocious luteal regression in the ewe was examined. Ewes were pre-treated on days 10 and 11 of the estrous cycle with either 750 mug E2 (total dose = 1.5 mg) in oil or with oil (O) alone. Treatment involved in delivery of 10 mug of synthetic LHRH/FSHRH in saline or an equal volume of saline only, administered at 2-h intervals beginning on day 12 of the estrous cycle and continuing through the succeeding 72 hours. During the period of LHRH administration, the serum LH patterns in the O-LHRH and E2-LHRH groups were characterized by rhythmic fluctuation, rising in response to LHRH and falling prior to the subsequent treatment injection. Throughout the course of the treatment period, the serum LH levels in the O-LHRH group were consistently higher than those in the E2-LHRH group. No increase in serum LH concentration was observed in the saline-treated animals. The mean luteal weight and mean luteal progesterone content at the end of the 72-h period were not significantly different between the O-S and E2-LHRH groups (543 +/- 88 vs. 455 +/- 126 mg and 13.1 +/- 6.2 vs 16.0 +/- 9.7 mug, respectively). Both luteal weight and progesterone content were increased (P less than .01) in the O-LHRH group (1089 +/- 87 mg and 47.5 +/- 3.1 mug) and significantly reduced (P less than .05) in the E2-S group (309 +/- 49 mg and 5.2 +/- 0.1 mug) compared with those of either the O-S group or the E2-LHRH group. Thus LHRH treatment increased mean luteal weight and mean luteal progesterone content while E2 pre-treatment depressed the same parameters. These data suggest that pulsatile administration of synthetic LHRH is able to elevate serum LH levels to an extent sufficient to counteract both natural luteolysis and premature luteal regression induced by E2 treatment.

Effects of indomethacin, piroxicam and selected prostanoids on gastric acid secretion by the rat isolated gastric mucosa.

The effects of the cyclo-oxygenase inhibitors indomethacin and piroxicam have been investigated on histamine- and dibutyryl cyclic AMP-induced acid secretion in the rat isolated gastric mucosa. The relative potencies of a number of prostanoids as inhibitors of histamine-induced acid secretion were determined in an attempt to classify the prostaglandin receptor mediating this response. Indomethacin (8 X 10(-9) - 2.7 X 10(-6) M) and piroxicam (3 X 10(-6) M) potentiated the secretory responses elicited by histamine. This effect might be due to inhibition of the biosynthesis of antisecretory prostanoids. Indomethacin (2.7 X 10(-6) M) and piroxicam (3 X 10(-6) M) also potentiated the secretory response to dibutyryl cyclic AMP, but since prostaglandin E2 (PGE2, 10(-5) M) did not inhibit this secretory response, the mechanism of the potentiation may differ from that of histamine. The potency of the thromboxane mimetic U-46619 as an inhibitor of histamine-induced acid secretion was markedly reduced in the presence of indomethacin, suggesting that U-46619 may release endogenous antisecretory prostanoids. In the presence of indomethacin (2.7 X 10(-6) M) all the prostanoids tested produced concentration-related inhibitions of histamine-induced gastric acid secretion. PGE-analogues were the most potent compounds, the rank order of potency being 16, 16 dimethyl PGE2 greater than PGE2 greater than PGF2 alpha greater than U-46619 greater than PGD2 greater than PGI2. This order of potency is very similar to that obtained in smooth muscle preparations containing 'EP' receptors, suggesting that this receptor type also mediates inhibition of histamine-induced acid secretion in the rat.

Further investigations into adenosine A1 receptor-mediated contraction in rat colonic muscularis mucosae and its augmentation by certain alkylxanthine antagonists.

1. The alkylxanthine antagonists, 8-phenyltheophylline (8-PT), 8-p-sulphophenyltheophylline (8-SPT) and 1,3,7-trimethylxanthine (caffeine) produced rightward displacements of contractile concentration-effect curves to 5'-N-ethylcarboxamidoadenosine (NECA) in rat isolated colonic muscularis mucosae (RCMM) with concentration-ratios consistent with adenosine receptor blockade. The non-xanthine antagonist, 9 fluro-2-(2-furyl)-5,6-dihydro [1,2,4] triazo to [1,5-c]-quinazin-imine (CGS15943A) also antagonized contractions to NECA with an affinity (pKB8.1-8.5) consistent with adenosine A1 receptor blockade. 2. In addition to producing rightward shifts of the concentration-response curves, the maximum contractions to 5'-N-ethylcarboxamidoadenosine (NECA) were also markedly increased in the presence of 8-PT (by 83 +/- 16% at 1 microM), 8-SPT (by 37 +/- 7% at 10 microM) and caffeine (by 45 +/- 5% at 100 microM) but were unaffected by CGS15943A (at 0.01 and 0.03 microM). 3. As with NECA, the maximum contractions to the adenosine A1 receptor agonists R-phenylisopropyladenosine (R-PIA) and N-[(1S, trans)-2-hydroxyclopentyl] adenosine (GR79236) were both antagonized and augmented by 8-PT. In addition, the contractions to NECA in the presence of 8-PT (1 microM) were inhibited by nanomolar concentrations of 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). 4. The non-selective phosphodiesterase (PDE) inhibitor, 3-isobutyl-1-methylxanthine (1 microM) produced a marked increase in the NECA maximum without producing a rightward shift in the NECA curve, whereas a higher concentration (10 microM) virtually abolished responses. The PDE type III inhibitor,milrinone (1 microM), the type IV inhibitor, rolipram (10 microM), and the type V PDE inhibitor, zaprinast(3 microM), were all without effect on NECA responses in RCMM.5. Partial inhibitions of contractions to NECA were produced by indomethacin (at 3 or 10 micro M) or piroxicam (at 3 microM). Responses to GR79236 were also partially inhibited by indomethacin. In the presence of indomethacin, 8-PT was still able to enhance markedly the maximum contractions obtained to NECA in RCMM.6. The present study has shown that certain alkylxanthine antagonists (but not the non-xanthineCGS15943A) produced a marked augmentation of adenosine Al receptor-mediated contractions inRCMM. The mechanism of this augmentation is, as yet, not known but is unlikely to result from inhibition of PDE. This study has also shown that adenosine Al receptor-induced contractions inRCMM are mediated, in part, via products of the cyclo-oxygenase pathway.

Investigation into the 5-hydroxytryptamine receptor mediating smooth muscle relaxation in the rat oesophagus.

1. An investigation has been made into the 5-hydroxytryptamine (5-HT) receptor mediating relaxation of rat oesophagus in preparations precontracted with carbachol. 2. In tissues treated with pargyline (100 microM) and in the presence of corticosterone (30 microM) and cocaine (30 microM) the potency of 5-HT and 5-methoxytyramine (5-MeOT) was not changed but the maximum response to these agonists was reduced. Thus there was no evidence of metabolism and/or uptake through an amine depleting mechanism. 3. The relaxant concentration-effect curves to 5-HT were shifted to the left in a concentration-related manner by isobutylmethylxanthine (1 and 10 microM), suggesting the involvement of adenosine 3':5'-cyclic monophosphate in these responses. 4. 5-HT produced concentration-related relaxations of rat oesophagus with an EC50 value of 0.24 microM. Several indole agonists were tested and the following rank order of potency of key agonists obtained: 5-HT greater than alpha-methyl-5-hydroxytryptamine = 5-carboxamidotryptamine (5-CT) greater than 5-MeOT. In contrast, 2-methyl-5-hydroxytryptamine, sumatriptan and 8-hydroxy-2-(di-n-propylamino) tetralin were weak or inactive. 5. The substituted benzamides, metoclopramide, cisapride, renzapride and R,S-zacopride acted as partial agonists, producing 60-70% of the 5-HT maximum. 6. The relaxation responses to 5-HT were neither inhibited by antagonists selective for 5-HT1 or 5-HT2 receptors nor by the 5-HT3 receptor antagonists, ondansetron, granisetron or MDL 72222. 7. The relaxation responses induced by 5-HT, 5-CT, 5-MeOT and renzapride were selectively inhibited by high concentrations of ICS 205-930 with pKB values of approximately 6. 8. The 5-HT receptor mediating relaxation in rat oesophagus cannot be designated 5-HT1, 5-HT2 or 5-HT3 under the current 5-HT classification, but the observed effects are consistent with stimulation of the putative 5-HT4 receptor.

Pharmacological basis for the induction of gastric carcinoid tumours in the rat by loxtidine, an insurmountable histamine H2-receptor blocking drug.

The very late occurrence of gastric carcinoids in a life-span carcinogenicity study with loxtidine in the rat might have resulted from continuous achlorhydria induced by this long-acting unsurmountable histamine H2-antagonist. The nature of the anti-secretory activity of loxtidine was compared with that of ranitidine on histamine-induced acid secretion in the perfused stomach preparation of the rat and in the rat isolated gastric mucosa preparation. Ranitidine and loxtidine had qualitatively different inhibitory effects on acid secretion, ranitidine being a competitive antagonist of histamine even at high concentrations, whereas the effect of loxtidine on both preparations was unsurmountable at relatively low concentrations. These results support the hypothesis that the late formation of gastric carcinoids in rats receiving loxtidine is a consequence of persistent achlorhydria caused by unsurmountable blockade of parietal cell H2-receptors.

Characterization of the adenosine receptor mediating contraction in rat colonic muscularis mucosae.

1. The objective of this study was to characterize the adenosine receptor mediating contraction in rat isolated colonic muscularis mucosae (RCMM). 2. Sequential additions of the adenosine receptor agonist 5'-N-ethylcarboxamidoadenosine (NECA; 0.01-10 microM) elicited reproducible, concentration-related contractions in RCMM. The effects of NECA were mimicked by the adenosine A1 receptor-selective agonists cyclopentyladenosine (CPA), R-phenylisopropyladenosine (R-PIA) and N-[1S, trans)2-hydroxycyclopentyl] adenosine (GR79236) and by S-PIA (the stereoisomer of R-PIA). The adenosine A2 agonists N-[(2-methylphenyl)methyl] adenosine (metrifudil) and 2-[p-(2-carboxyethyl)phenethylamine]-5'-N-ethylcarboxamidoadenosine (CGS21680) also produced contractions in RCMM but were 54 and 165 times less potent respectively than NECA. The rank order of agonist potency for contraction of RCMM was CPA > or = GR79236 = R-PIA > or = NECA > > S-PIA = metrifudil > CGS21680, which is identical to that reported for the inhibition of spontaneous rate in rat isolated right atria and inhibition of lipolysis in rat isolated adipocytes by these same agonists. 3. R-PIA, S-PIA and metrifudil behaved as partial agonists in RCMM. 4. The adenosine A1 receptor-selective antagonist 8-cyclopentyl-1,3- dipropylxanthine (DPCPX) inhibited the contractions produced by all the adenosine agonists tested, with pKB values between 9.2 and 9.5. The non-selective adenosine antagonist 8-phenyltheophylline (8-PT) antagonized the effects of NECA but also markedly potentiated (by 93.0 +/- 10.2% at 3 microM) the maximum contractile response to NECA in RCMM. Neither 8-PT (3 microM) nor DPCPX (0.1 microM) had any effect on the contractions produced by carbachol. 5. The contractile responses to NECA in RCMM were not affected by atropine (1 microM), tetrodotoxin(0.3 microM) or the P2 antagonist, suramin (100 microM).6. The present study confirms that contractions to adenosine agonists in the RCMM are mediated via adenosine Al receptors.

Regulation of the immunoglobulin G1 receptor: effect of prolactin on in vivo expression of the bovine mammary immunoglobulin G1 receptor.

Induction of colostrogenesis in non-pregnant cows was used to evaluate the relationship between prolactin (PRL) and mammary immunoglobulin G1 (IgG1) receptor expression. Six of eleven non-pregnant, non-lactating Holstein cattle responded to a standard lactation induction protocol by development of elevated IgG1 concentrations in mammary secretions. In order to increase the diversity in PRL concentrations, two of the six cattle were treated with bromocriptine, and two others were treated with recombinant bovine PRL. Serum alpha-lactalbumin, serum PRL and mammary secretion IgG1 concentrations were measured throughout the experiment. Biopsies of mammary tissue were collected after induction of lactation, and after treatments to alter serum PRL. Immunohistochemistry was used to evaluate IgG1 receptor expression. Administration of recombinant bovine (rbPRL) was associated with increased lactogenic activity, decreased secretion IgG1 concentrations, and decreased IgG1 receptor expression. Decreased serum PRL, due to bromocriptine, was associated with decreased lactogenic activity and maintenance of IgG1 receptor expression. Results of this experiment are consistent with an effect of PRL in decreasing the expression of the bovine mammary IgG1 receptor at the onset of lactogenesis.

Specific binding of a potent LHRH agonist in rat testis.

High affinity binding sites for the potent LHRH agonist [125I][D-Ser(TBU)6, des-Gly-NH2(10)]LHRH ethylamide are present in dissociated rat testicular interstitial cells, a preparation rich in Leydig cells. The iodinated LHRH agonist binds to a single class of high affinity sites at a KD value of 0.12 nM and the number of binding sites is approx. 2500 per interstitial cell. A close correlation is observed between the potency of representative LHRH agonist to stimulate LH release in anterior pituitary cells in culture and their affinity for the testicular binding sites. The presence of specific LHRH receptors in an enriched population of Leydig cells suggests that these receptors could play a role, not only in the antifertility effects of LHRH agonists, but also in the physiological control of testicular functions.

Chronic administration of H2-antagonists does not alter gastric secretory responses to histamine, or the antisecretory activity of sufotidine.

Gastric secretory responses to histamine were investigated in anaesthetized dogs following treatment with oral ranitidine at 5 mg/kg twice daily for 358 weeks, and in isolated gastric mucosae from mice receiving sufotidine 240-280 mg.kg/day for 15 months. In neither study were there any significant differences between the acid secretory dose-response curves to histamine in control and test animals. The antisecretory activity of oral sufotidine (1 mg/kg) against histamine-induced acid secretion in the Heidenhain pouch dog was unaltered by twice daily dosing with sufotidine for 14 days. These studies on the effects of H2-antagonists on histamine-stimulated acid secretion found no evidence for development of direct tolerance at the parietal H2-receptor level.

Reproductive and endocrine changes in ewes actively immunized against estrogens and androgens.

Twenty ewes were divided equally into two treatments to be actively immunized simultaneously against estrogen and androgen conjugated to ovalbumin (steroid immunized) or immunized against ovalbumin alone (control) to determine the effects of steroid immunization on reproduction and pituitary function. Ewes were immunized at week 0, 3 and 5 of the 93-week study and exposed to a fertile ram for two breeding seasons (weeks 7-25 and weeks 60-67). Steroid immunization reduced the proportion of ewes which exhibited estrus during the first breeding season when antibody activity against steroids was highest and reduced the proportion of ewes lambing after both breeding seasons. Ovulation rate, observed only during the second breeding season, was two times higher in steroid immunized ewes, but no increase in percent lamb crop per ewe lambing was observed. Pituitary concentrations of immunoactive LH (determined by radioimmunoassay), bioactive LH (determined by stimulation of mouse Leydig cell production of testosterone) and receptors for LHRH were increased by steroid immunization. These changes in pituitary LH resulted in a decreased ratio of bioactive to immunoactive LH observed in the pituitary and a decrease in circulating concentrations of bioactive LH. It was concluded from this study that simultaneous active immunization against estrogens and androgens increases ovulation; however, estrus and lambing rates may be reduced when antibody activity is high. Ovulation rates were increased despite the fact that no change in FSH was observed and the biopotency of LH was decreased in steroid immunized ewes.

Reproductive and endocrine changes in ewes actively immunized against luteinizing hormone.

The objective of this study was to examine reproductive and endocrine changes in ewes actively immunized against a conjugation of luteinizing hormone (LH) and ovalbumin (LH immunized n = 10) or ovalbumin alone (control; n = 10). Ewes were immunized at weeks 0, 3 and 5 of the 93-week study and exposed to a fertile ram during weeks 7-25 (first breeding season) and weeks 60-67 (second breeding season). Immunization against LH abolished estrus and prevented pregnancy in all but one LH immunized ewe during the two breeding seasons. Uterine weights of LH immunized ewes were lighter than those observed in control ewes while ovarian weights did not differ between the treatment groups. No differences were observed between basal circulating concentrations of LH measured by an in vitro bioassay for the two treatment groups. However, serum concentrations of immunological follicle stimulating hormone, and the number of medium sized ovarian follicles (4-6 mm) were increased in LH immunized ewes. It was concluded from this study that immunization against LH prevents ovulation presumably by blocking the preovulatory surge of LH and not altering basal concentration of LH.

Response to a growth hormone-releasing hormone analog in heifers treated with recombinant growth hormone.

Sixteen pregnant Holstein heifers (430kg) were used to determine the effect of long-term administration of a bovine growth hormone (bGH) made by recombinant DNA technology on the ability of a bolus injection of a growth hormone-releasing hormone analog (Ac-His-1, D-Ala-2, Nle-27, GHRH(1-29 NH2) to increase serum GH. Eight heifers received a daily intramuscular injection of bGH (50 mg/day) for 5 months while the other half received a daily injection of physiological saline (control) over the same period. On the last day of bGH treatment and 1, 5, 10 and 25 days after the cessation of bGH treatment, five heifers from each group were challenged with GHRH analog and the response to this releasing hormone analog was measured. Basal GH concentrations were elevated on the last day of treatment in bGH-treated heifers and declined to concentrations similar to control heifers by 1 day after cessation of treatment. Response to GHRH analog was impaired by bGH during the last day of treatment and one day later. Responsiveness returned to a level similar to controls by 5 days after the end of bGH treatment. Response to GHRH analog was lessened during the period of bGH treatment but there were no long term effects on the animals' ability to respond to the releasing hormone.

Effects of estradiol on secretion of LH, hypothalamic function and testicular development in bull calves.

Two experiments were conducted in order to determine the effects of estradiol (E2) on the development of the hypothalamic-pituitary-testicular axis in bull calves. In experiment 1, calves were assigned randomly to one of the following groups: 1) intact, 2) intact E2-treated, 3) castrated, or 4) castrated E2-treated. Treatments began when the calves were 7.5 wk of age and continued for 16.5 wk. Samples of blood were collected once a week from 3 to 14 wk of age and every 10 min for 6 hr at 8, 12 and 16 wk of age. Concentrations of E2 in plasma decreased between 3 and 4 wk of age and were further reduced by castration. Maximum concentrations of E2 (24.3 pg/ml) were observed 72 hr after insertion of E2 implants, however, plasma E2 stabilized at 5.9 pg/ml by 2 wk after insertion of E2 implants. Treatment with E2 eliminated the pulsatile secretion of LH in intact and castrated calves and retarded testicular growth. In experiment 2, calves were assigned to a control (n = 4) or E2-treated (n = 6) group. Implants of E2 were inserted at 7.5 wk of age. At 24 wk of age, calves were bled and then sacrificed to collect hypothalamic and pituitary tissues. Age-related changes in testicular weight and secretion of LH were blocked by E2. Neither the morphology nor the intensity of immunostaining of GnRH nerve cell bodies in the preoptic area (POA) were affected by E2. However, the density of GnRH fibers and beads in the stalk median eminence (SME), and concentrations of pituitary GnRH receptors were greater (P less than .01) in E2-treated compared to control calves. In addition, concentrations of norepinephrine (NE) in the SME were lower in E2-treated calves when compared to controls. Based on these observations, it is concluded that administration of E2 at 7.5 wk of age causes profound alterations in hypothalamic function including, changes in metabolism of NE and suppression of GnRH release.

Adenohypophyseal receptors for LHRH, pituitary content of gonadotropins and plasma concentrations of LH in cyclic, pregnant and postpartum beef cows.

Adenohypophyseal concentrations of LHRH receptors, pituitary content of LH and FSH, and plasma concentrations of LH were determined in thirty Hereford, Angus or Hereford-Angus heifers that were randomly assigned by breed and weight to five periods including day 3 of the estrous cycle (CY), pregnant day 120 (P120), 200 (P200), 275 (P275), or day 2 postpartum (PP). Jugular blood samples were collected at 10-min intervals for 8 hr from all cows. Within 2 hr after completion of blood sampling, animals were slaughtered and the pituitary gland frozen at -196 C. LH pulse frequency/8 hr was reduced (P less than .05) during gestation (.5, .2, and 1.5 +/- .5/8 hr, for P120, P200, and P275, respectively) and PP (.5 +/- .5/8 hr) compared to CY (7.8 +/- .5/8 hr). Frequency of LH pulses/8 hr was not different (P greater than .1) among P120, P200 or PP periods but was different (P less than .05) between P200 and P275. There were no differences in LH pulse height (P greater than .1) among periods; however, pulse amplitude was greatest (P less than .05) at P120 (1.3 +/- .2 ng/ml) and lowest between P200 and PP (.6 to .8 +/- .2 ng/ml). Baseline concentrations of plasma LH did not differ (P greater than .1) among P and PP periods (.3 +/- .1 ng/ml), but were lower (P less than .05) than in CY animals (.7 +/- .1 ng/ml). Concentration of adenohypophyseal LHRH receptors was approximately two-fold greater (P less than .05) at P120 (25.85 +/- 2.2 fmol/mg) than at all other periods (9.5 to 14.9 +/- 2.2 fmol/mg).(ABSTRACT TRUNCATED AT 250 WORDS)

Synchronization of estrus in yearling beef heifers with melengestrol acetate and prostaglandin F(2alpha).

This study was designed to test the efficacy of melengestrol acetate (MGA) in combination with prostaglandin F(2alpha) (PGF(2alpha)) in synchronizing estrus in cyclic and noncyclic heifers. One hundred thirty-one cyclic and prepubertal crossbred heifers were randomly assigned to three treatment groups: Controls (n = 43); MGA (0.5 mg/d for 7 d) and PGF(2alpha) (25 mg i.m. on Day 7; n = 44); and PGF(2alpha) (25 mg i.m. on Day 7; n = 44). Observations for estrus were made at 6-n intervals throughout the 7-d treatment period followed by a 34-d artificial insemination breeding season. A greater percentage (P < 0.05) of MGA-PGF(2alpha) noncyclic heifers showed behavioral estrus (91%) than did Control (67%) or PGF(2alpha) heifers (61%) during the 34-d artificial insemination period. There was no difference (P > 0.05) between synchronization rates of the MGA-PGF(2alpha) heifers and PGF(2alpha) heifers 7 d after PGF(2alpha) administration. The percentage of control animals in estrus during the first 25 d of the breeding season did non differ from the synchronized rates of MGA-PGF(2alpha) and PGF(2alpha) heifers (P > 0.05). Conception rates (heifers pregnant/heifers inseminated) did not differ (P > 0.05) for cyclic or prepubertal heifers among Control, MGA-PGF(2alpha) or PGF(2alpha) heifers. Though conception rates did not differ, there was a trend toward lowered conception rates in MGA-PGF(2alpha) heifers.

Elevated inhibin concentration in the follicular fluid of dairy cows with chronic cystic ovarian disease.

This study compared serum and follicular fluid inhibin and gonadotropin profiles between chronic cystic ovarian diseased (CCOD) and normal cyclic dairy cows. Blood samples and follicular fluid were collected from CCOD cows (n=15) and cyclic cows in the follicular phase of the estrous cycle (control, n=6) and analyzed for inhibin, follicle stimulating hormone (FSH) and luteinizing hormone (LH) concentrations. There was a significant increase in inhibin and a decrease in FSH and LH concentrations in the follicular fluid of CCOD cows compared with those of cyclic cows (P < 0.05). Mean serum inhibin, FSH and LH concentrations between CCOD and cyclic cows were not differnt (P > 0.05), however, there was a tendency for serum inhibin to be higher and FSH to be lower in CCOD cows compared to cyclic animals (P < 0.1). The FSH pulse frequency also was lower in CCOD cows than in cyclic cows (P < 0.05). These data suggest that increased production of inhibin from cystic follicles of CCOD cows alters pituitary FSH secretion and subsequently reduces the concentration of FSH in follicular fluid. As a result, decreased FSH stimulation at the ovarian level could ultimately lead to the reduction in follicular LH and FSH receptor concentrations, resulting in abnormal follicular steroidogenesis in CCOD dairy cows.