Biomedical subjects

G E Moss

Publications and source records attributed to G E Moss.

At least 37 records · Page 2Linked to original sources

PubMed1998

Regulation of amounts of mRNA for GnRH receptors by estradiol and progesterone in sheep.

Expression of GnRH receptors increases prior to the onset of the preovulatory surge of LH in sheep. Two experiments were conducted to investigate the interactions of progesterone (P) and estradiol (E) on amounts of mRNA for GnRH receptors and the number of receptors for GnRH. The first study was designed as a 2 x 2 factorial arrangement of treatments to investigate effects of removal of P and the presence of E. Ewes that had been ovariectomized (OVX) for at least 4 wk received one silastic implant containing E and two silastic implants containing P for 6 d to mimic concentrations of these steroids during the luteal phase of the estrous cycle. Anterior pituitary glands were collected (n = 4 animals/group): 1. Prior to implant removal and 12 h after removal of: 2. P only. 3. E only. 4. P and E. Regardless of whether or not E was present, amounts of mRNA for GnRH receptors (P = 0.87) and number of GnRH receptors (P = 0.43) were not different within 12 h after removal of P. In the second experiment, ewes were OVX on d10-12 of the estrous cycle (d0 = estrus), and immediately received silastic implants containing E and P as described above. Anterior pituitary glands were collected on d12 of the estrous cycle (n = 5), prior to implant removal (n = 5), and from the remaining ewes 24 h after removal of P only (n = 7) or removal of P and E (n = 6). Relative amounts of mRNA for GnRH receptors and the number of GnRH receptors were similar (P > 0.05) on d12 of the estrous cycle and prior to implant removal. Removal of both P and E did not affect (P > 0.05) amounts of GnRH receptor mRNA or number of GnRH receptors. However, the removal of P and the presence of E increased (P < 0.05) amounts of mRNA for GnRH receptors, but did not affect (P > 0.05) the number of GnRH receptors. We conclude that increased amounts of GnRH receptor mRNA require the removal of P and the presence of E.

Animals↗

PubMed1998

Estradiol and gonadotropin-releasing hormone (GnRH) interact to increase GnRH receptor expression in ovariectomized ewes after hypothalamic-pituitary disconnection.

Gonadotropin-releasing hormone (GnRH) receptor expression is regulated by estradiol and GnRH itself. The objective of this experiment was to determine the extent to which low levels of estradiol, similar to those observed during the transition from the luteal to the follicular phase of the estrous cycle, and GnRH interact to regulate expression of GnRH receptors and GnRH receptor mRNA. Ewes were ovariectomized (OVX) at least 2 wk prior to initiation of the experiment, and the pituitary gland was surgically disconnected from the hypothalamus to remove ovarian and hypothalamic inputs to the pituitary. Within 24 h after hypothalamic-pituitary disconnection, ewes received pulses of GnRH (250 ng/pulse) every 2 h for 6 d. At the end of 6 d, ewes were randomly assigned to treatments in a 2 x 2 factorial arrangement as follows: half of the animals received a single estradiol implant and half received an empty implant (placebo). At the same time, animals also received one of the following treatments: (1) saline or (2) GnRH (100 ng/pulse/2 h). Additionally, one group of ewes was ovariectomized, but not subjected to hypothalamic-pituitary disconnection (OVX controls). Blood samples were collected 15 min prior to each pulse of GnRH or saline and at 15-min intervals for 1 h after each pulse until tissues were collected and concentrations of luteinizing hormone (LH) were determined. Anterior pituitaries were collected 24 h after implant insertion to quantitate steady-state amounts of GnRH receptor mRNA and numbers of GnRH receptors. Mean LH was greatest in ovariectomized control ewes compared to all other treatments (p < 0.05). Mean LH and LH pulse amplitude in the placebo and GnRH-treated group most closely mimicked LH secretion in ovariectomized control animals. Mean LH and LH pulse amplitude were similar between both GnRH-treated groups (p < 0.05). Mean LH and LH pulse amplitude were significantly lower in all animals treated with saline compared to OVX controls (p < 0.05). Treatment with an estradiol implant and pulsatile GnRH increased (p < 0.05) relative amounts of GnRH receptor mRNA and the number of GnRH receptors compared to all other treatments. There were no differences in GnRH receptor expression between the remaining treatment groups (p > 0.05). Therefore, in OVX ewes after hypothalamic-pituitary disconnection, low levels of estradiol and GnRH are required to increase GnRH receptor mRNA and GnRH receptor numbers. Since we only observed an increase in GnRH receptor expression in the presence of both estradiol and GnRH, we conclude that there is a synergistic interaction between these two hormones in the regulation of GnRH receptor expression.

Animals↗

PubMed1998

Regulation of ovine GnRH receptor gene expression by progesterone and oestradiol.

The objective of this study was to determine whether progesterone prevents the stimulatory effects of oestradiol on GnRH receptor gene expression. In Expt 1, ewes were treated during the luteal phase (days 10-12 of the oestrous cycle) with either one or five subcutaneous implants containing oestradiol (n = 6 per group). Control ewes received no treatment (n = 6). Anterior pituitary glands were collected 16 h after treatment with oestradiol. Steady-state amounts of GnRH receptor mRNA were similar among all three treatment groups despite increased circulating concentrations of oestradiol in implanted ewes at the time of pituitary collection (4.3 +/- 0.6 and 24.7 +/- 2.6 pg ml-1 in ewes treated with one or five implants, respectively, compared with 0.5 pg ml-1 in controls). Experiment 2 was designed to determine whether progesterone was the ovarian factor preventing the stimulatory effects of oestradiol on expression of the GnRH receptor gene in Expt 1. Twenty-five ewes were ovariectomized on day 6 or day 7 of the oestrous cycle and assigned to one of five treatment groups (n = 5 per group). Control ewes received no further treatment. Endogenous luteal phase concentrations of progesterone were replaced in three groups of ewes at the time of ovariectomy via intravaginal implants. Three days after ovariectomy, one group of progesterone-treated ewes received one oestradiol implant, while another group of progesterone-treated ewes received five oestradiol implants. An additional group was treated with five oestradiol implants only, and anterior pituitary glands were collected from all ewes 16 h later. Compared with untreated ovariectomized ewes, treatment with progesterone alone did not affect amounts of GnRH receptor mRNA. In ewes treated with progesterone and either one or five oestradiol implants, steady-state amounts of GnRH receptor mRNA were increased twofold (P < 0.01). Treatment with oestradiol in the absence of progesterone increased amounts of GnRH receptor mRNA threefold (P < 0.001). These results provide evidence that the stimulatory effects of oestradiol on the expression of the GnRH receptor gene are prevented during the natural luteal phase in ewes. However, progesterone does not appear to act independently to mediate this effect.

Analysis of Variance↗

PubMed1998

Steady-state concentrations of mRNA encoding two inhibitors of protein kinase C in ovine luteal tissue.

Prostaglandin F2 alpha (PGF2 alpha) decreases secretion of progesterone from the corpus luteum in domestic ruminants. However, it is less effective during the early part of the oestrous cycle (Louis et al., 1973) and at the time of maternal recognition of pregnancy (Silvia and Niswender, 1984; Lacroix and Kann, 1986). Decreased luteal responsiveness may be due to failure of PGF2 alpha to activate fully its normal second messenger system, protein kinase C (PKC). Alternatively, increased resistance of the corpus luteum to PGF2 alpha might be attributable to greater concentrations of recently identified biological inhibitors of PKC. These possibilities were addressed by measuring steady-state concentrations of mRNA encoding PGF2 alpha receptor and two inhibitors of PKC, protein kinase C inhibitor-1 (PKCI-1) and kinase C inhibitor protein-1 (KCIP-1, brain 14-3-3 protein), in corpora lutea collected from ewes on days 4, 10 and 15 of the oestrous cycle (n = 5 per day) and day 15 of pregnancy (n = 7). There were no differences in mean concentrations of mRNA encoding PGF2 alpha receptor among the groups. However, concentrations of mRNA encoding both inhibitors of PKC were higher (P < 0.01) on day 4 of the oestrous cycle compared with the other groups. Treatment of ewes with a luteolytic dose of PGF2 alpha, which activates PKC, did not change concentrations of mRNA encoding either PKCI-1 or KCIP-I up to 24 h later. Luteal expression of mRNA encoding the PKC inhibitors and PGF2 alpha receptor was also examined in ewes treated with oestradiol in vivo for 16 h in the midluteal phase. High concentrations of oestradiol in serum (20 and 70 pg ml-1) did not influence quantities of any of the mRNAs examined. Therefore, an increase in PKC inhibitors may be involved in resistance of the corpus luteum to PGF2 alpha during the early part of the oestrous cycle but does not appear to mediate the increased resistance of the corpus luteum to PGF2 alpha during maternal recognition of pregnancy. Neither PGF2 alpha nor oestradiol affected steady-state concentrations of mRNAs encoding PKCI-1 or KCIP-I.

14-3-3 Proteins↗

PubMed1997

Induction of the heat shock response and translational thermotolerance in day 15 ovine trophectoderm.

The objectives of this study were to determine the ability of trophectoderm from preimplantation ovine embryos to synthesize hsp70 in response to heat shock and to identify conditions which induce translational thermotolerance in this tissue. Day 15 embryos were collected, and proteins synthesized in 1.5-mm sections of trophectoderm were radioactively labeled with (35)S-methionine. One-dimensional SDS-PAGE gels, two-dimensional gel electrophoresis and Western blots were utilized to characterize the heat shock response and to examine the induction of translational thermotolerance. Increased synthesis of the 70 kDa heat shock proteins and a protein with an approximate molecular weight of 15 to 20 kDa was observed with heat shock (> or = 42 degrees C). Total protein synthesis decreased (P < 0.05) with increased intensity of heat shock. At 45 degrees C, protein synthesis was suppressed with little or no synthesis of all proteins including hsp70. Recovery of protein synthesis following a severe heat shock (45 degrees C for 20 min) occurred faster (P < 0.05) in trophectoderm pretreated with a mild heat shock (42 degrees C for 30 min) than trophectoderm not pretreated with mild heat. In summary, trophoblastic tissue obtained from ovine embryos exhibit the characteristic "heatshock" response similar to that described for other mammalian systems. In addition, a sublethal heat shock induced the ability of the tissue to resume protein synthesis following severe heat stress. Since maintaining protein synthesis is crucial to embryonic survival, manipulation of the heat-shock response may provide a method to enhance embryonic survival.

Journal Article↗

PubMed1996

Effects of porcine relaxin on induced parturition in beef heifers.

Two-year-old crossbred beef heifers were used to test the effects of porcine relaxin (pRelaxin) alone, or in combination with dexamethasone, on the induction of parturition, the incidence of dystocia, and retained placentas. Effects of treatment on pelvic area, postpartum interval, milk production, colostrum quality, calf birth weight, calf vigor, and calf performance were also evaluated. On Day 275 of gestation, heifers from two fetal-sire groups were randomly assigned to one of four groups in a 2 x 2 factorial design and received; no treatment (controls, n = 19), 20 mg of dexamethasone intramuscularly (im) (n = 22), 5 mg of pRelaxin (3,000 U/mg) im (n = 19), or 20 mg of dexamethasone plus 5 mg of pRelaxin (n = 17). Length of gestation (in days) was less (P < 0.05) in heifers treated with dexamethasone (279.8 +/- 1.0) than in controls (286.6 +/- 0.9), but was not influenced (P > 0.05) by treatment with pRelaxin. The incidence of retained placentas in heifers treated only with dexamethasone (27.3%) was not reduced by concomitant treatment with pRelaxin (35.3%). Retained placentas were not observed in any control heifers and in only one heifer (5.2%) treated solely with pRelaxin. Ease of calving (1 = unassisted, 5 = abnormal presentation) was not influenced by treatment (P > 0.05), even though birth weights (in kilograms) of calves from heifers treated with dexamethasone (36.4 +/- 0.8) were less (P < .01) than those of calves from nondexa-methasone-treated heifers (39.2 +/- 0.8). Dexamethasone tended to reduce (P < 0.07) calf vigor (1 = healthy and strong, 5 = dead on arrival; 1.48 +/- 0.11 vs. 1.18 +/- 0.11), but was not (P > 0.05) influenced by pRelaxin. The duration of the postpartum anestrous interval (73.1 +/- 1.8 d across groups) and pelvic areas following treatment and parturition were not influenced (P > 0.05) by dexamethasone or pRelaxin. Although determinants of colostrum quality (P < 0.01) and quantity (P < 0.08) of milk produced were influenced by dexamethasone, adjusted 205-d weights of calves did not differ (P > 0.05) among groups. In conclusion, treatment with pRelaxin alone failed to induce parturition or, when combined with dexamethasone, to reduce the incidence of retained placentas.

Animals↗

PubMed1995

A comparison of LH secretion and brain estradiol receptors in heterosexual and homosexual rams and female sheep.

This study examined endocrine components of sexual orientation of male sheep. Sexual orientation of adult rams was identified through standardized sexual performance tests. Four rams that copulated with ewes, four rams that never mounted females and copulated with males, and eight ewes were used in the experiments. Exogenous estradiol benzoate (50 micrograms, i.m.) stimulated (P < .05) a preovulatory-like LH surge 16-22 hr after administration to females. Estradiol did not (P > .05) affect LH release of heterosexual or homosexual rams. Thirty days after the estradiol challenge, sheep were euthanized and areas of the amygdala (AMY), hypothalamus (HYP), anterior pituitary (AP), and preoptic area (POA) of the hypothalamus were collected. Occupied and unoccupied content of estradiol receptors (ER) was determined. The content of ER in the amygdala of both homosexual rams and ewes was similar, but less than (P < .05) the content of ER in heterosexual rams. The ER content measured in other brain regions did not differ by sex or orientation. In summary, results from these data show that the preovulatory LH surge mechanism that is a characteristic of the female does not occur in either homosexual or heterosexual rams. Conversely, the ER content of the AMY of homosexual rams is similar to that of ewes and differs from the heterosexual male. Differences in ER content between heterosexual and homosexual rams imply that the amygdala serves as a link for input from potential mates. These data suggest that the amygdala not only plays a role in sexual behavior but may be involved in sexual orientation of rams.

Animals↗

PubMed1995

Influence of fetal death and fetectomy on gestation and the initiation of parturition in the ewe.

Depressed function of the fetal hypothalamic-pituitary-adrenal axis results in prolonged gestation, and fetal death causes premature parturition. The objective of this experiment was to determine effects of death in utero of a sibling, or its removal, on the duration of gestation and concomitant changes in maternal serum concentrations of oestradiol (E) and progesterone (P). Ovine placental lactogen (oPL) was also determined as an index of placental viability. Blood samples were collected in the morning, beginning 3 days prior to surgery on Day 115 +/- 3 of gestation and continuing daily until 3 days post partum. Surgeries were performed via mid-ventral laparotomy to induce fetal death or to remove the fetus. Fetal death was induced by ligating the umbilicus. Duration of gestation was similar (P > 0.05) in control (C, n = 6) and sham-operated (S, n = 3) ewes (148 +/- 1.0 and 148.6 +/- 0.7 days, respectively). On the day of parturition, concentrations of P, E and oPL were 5.2 +/- 1.9 ng mL-1, 135 +/- 22 pg mL-1 and 153 +/- 54 ng mL-1, respectively, in ewes from combined C and S groups. Total fetectomy (n = 3) resulted in a rapid decrease (P < 0.05) in maternal serum concentrations of P, E, and oPL. Ligation of the umbilicus of both fetus(es) in utero (n = 4) induced fetal death, decreased (P < 0.05) length of gestation to 118.8 +/- 1.8 days and decreased (P < 0.05) serum concentrations of P and oPL prior to parturition and oPL on the day of parturition. In addition, maternal serum concentrations of E failed (P > 0.05) to increase at parturition. Length of gestation and concentrations of P, E and oPL at parturition were not affected (P > 0.05) by removal of one fetus when its sibling was undisturbed (n = 4) compared to control ewes. In contrast, death of one fetus with its sibling undisturbed (n = 5) decreased (P < 0.01) length of gestation (139.2 +/- 2.8 days), but did not affect P, E and oPL (P > 0.05) on all days tested. In conclusion, death of one fetus reduced the duration of gestation, but changes prepartum in maternal serum concentrations of P and oPL were similar to ewes delivering only live fetuses. The increase in maternal concentrations of E that normally occur at parturition was absent in ewes giving birth to only dead fetuses and, therefore, was not a prerequisite to parturition.

Abortion, Veterinary↗

PubMed1995

Effect of acute glucose antagonism on hypophyseal hormones and concentrations of insulin-like growth factor (IGF)-I and IGF-binding proteins in serum, anterior pituitary, and hypothalamus of ewes.

The objectives of this study were to determine whether 2-deoxy-D-glucose (2DG), a metabolic inhibitor of glucose, 1) differentially affected release of LH, growth hormone (GH) and prolactin (PRL); 2) decreased anterior pituitary response to GnRH; and 3) altered concentrations of insulin-like growth factor I (IGF-I) and intensity of 125I-IGF-I binding to proteins (IGFBPs) in serum, anterior pituitary (AP), and hypothalamus (preoptic are [POA], medial basal hypothalamus [MBH], and stalk median eminence [SME]). In trial 1, mature, ovariectomized, crossbred ewes were treated with a single s.c. injection of either saline (n = 5) or 2DG (n = 5; 0.1 g/kg BW). Administration of 2DG suppressed (p < 0.05) concentrations of LH but did not affect (p > 0.05) serum concentrations of GH or PRL. In trial 2, ovariectomized ewes received injections of saline or 2DG, and each group was subdivided to receive either saline or GnRH analog (5 micrograms; Des-Gly10-[D-Ala6]) 2 h later. Administration of 2DG suppressed (p < 0.05) serum concentrations of LH before administration of GnRH; however, release of LH in response to exogenous GnRH was greater (p < 0.05) in ewes that received 2DG. Treatment with 2DG did not affect (p > 0.05) concentrations of FSH before or after administration of GnRH. In trial 3, 10 ovariectomized ewes were slaughtered 2 h after treatment with saline (n = 5) or 2DG (n = 5). Serum and tissues collected at slaughter (AP, MBH, SME, and POA) were analyzed for concentrations of IGF-I by RIA and intensity of 125I-IGF-I binding to proteins by ligand blotting.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

Estradiol increases amounts of messenger ribonucleic acid for gonadotropin-releasing hormone receptors in sheep.

Two experiments were conducted simultaneously to investigate regulation of amounts of mRNA for GnRH receptors during the periovulatory period in sheep. In the first experiment, amounts of mRNA for GnRH receptors were measured before and after preovulatory surge of LH following regression of the CL by prostaglandin F2 alpha(PGF2 alpha). So that the time of the preovulatory surge of LH could be accurately predicted, ewes received two injections of PGF2 alpha on Day 14 of the estrous cycle. Anterior pituitary glands were collected from 5 control ewes on Day 14 of the estrous cycle (0 h after PGF2 alpha) and at 48, 72, and 96 h after PGF2 alpha (5 ewes per group). The second experiment was conducted to investigate the effects of 17 beta-estradiol on amounts of mRNA for GnRH receptors. On Day 14 of the estrous cycle, 20 ewes were ovariectomized (OVX); 15 of these ewes received estradiol implants when they were OVX (OVXEI). Sixteen hours after OVX, anterior pituitary glands were collected from 5 OVX and 5 OVXEI ewes, and the remaining OVXEI ewes received an i.m. injection of estradiol (25 micrograms in corn oil; OVXEI + E) to induce a preovulatory-like surge of LH. Anterior pituitary glands were collected from OVXEI + E ewes 18 or 54 h after injection of estradiol (n = 5 per group). Half of each anterior pituitary gland was used to measure the number of GnRH receptors. Poly(A)+ RNA was isolated from the remaining half of each anterior pituitary gland, applied to slot blots, and hybridized with a radioactive cDNA probe encoding the ovine GnRH receptor.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

Insulin-like growth factor-I (IGF-I) and IGF-binding proteins in bovine sera and pituitaries at different stages of the estrous cycle.

The objective of this study was to determine whether concentrations of hypophyseal and serum insulin-like growth factor-I (IGF-I) and binding activities of serum and hypophyseal IGF-binding proteins (IGFBPs) differ with stage of the estrous cycle in mature beef cows. Cows were assigned to the following stages of the estrous cycle based on serum concentrations of progesterone (P4) and ovarian structures at death: days 1-5 (day 0 = estrus; n = 18), days 6-10 (n = 24), days 11-16 (n = 39), and days 17-21 (n = 7). Serum samples collected at death and anterior pituitary homogenates were analyzed for IGF-I, LH, and FSH by RIA. Serum and pituitary IGFBPs were evaluated by ligand and immunoblot analyses. Serum samples contained IGFBP activity at 44 and 40 kilodaltons (kDa; IGFBP-3), 34 kDa (IGFBP-2), 30 kDa (IGFBP-5), 28 kDa, and 24 kDa. The intensity of binding by the different sized proteins in serum remained constant throughout the estrous cycle. IGFBPs detected in anterior pituitaries included a 36-/40-kDa doublet (IGFBP-3), a 32-kDa protein (IGFBP-2), and a 29-kDa IGFBP (IGFBP-5). The intensity of [125I]IGF-I binding to IGFBPs was greater (P < 0.05) during days 11-16 than days 1-5 or days 6-10 of the estrous cycle and was intermediate (P > 0.05) during days 17-21. Serum concentrations of LH were low (< 0.4 ng/ml) during days 1-16 of the estrous cycle, but increased (P < 0.05) approximately 3- to 4-fold during days 17-21. LH concentrations in the anterior pituitary increased (P < 0.05) from the postovulatory period (548 +/- 52 micrograms/g; days 1-5) to the late luteal phase (791 +/- 39 micrograms/g; days 11-16) and were intermediate (P > 0.05) during the preovulatory phase (707 +/- 85 micrograms/g; days 17-21). Concentrations of IGF-I and FSH in the anterior pituitary and serum did not differ (P > 0.05) by stage of the estrous cycle. A positive correlation among the different IGFBPs in the pituitary (P < 0.01) and between each pituitary IGFBP and serum P4 (P < 0.05) existed. The significance (P < 0.0002) of the correlation between pituitary IGFBP-3 and P4 was greater (P < 0.02) than that between the other pituitary IGFBPs and P4. In summary, IGFBP activity in the anterior pituitary, but not that in serum, changed with stage of the estrous cycle in association with serum concentrations of P4. The absence of similar changes in serum or anterior pituitary concentrations of IGF-I supports the hypothesis that IGFBPs may regulate the actions of IGF-I on gonadotropin release from the anterior pituitary gland during the estrous cycle of beef cattle.

Animals↗

PubMed1994

Posttranslational processing of progonadotropin-releasing hormone (proGnRH) in ewes.

Gonadotropin-releasing hormone (GnRH) is released from hypothalamic neurons into the hypophyseal-portal blood system following enzymatic cleavage of the decapeptide from a large precursor (proGnRH) molecule. The purpose of this study was to determine whether the ability of GnRH-producing neurons to synthesize and/or process proGnRH differed during physiological states associated with a suppressed and enhanced release of GnRH in ewes. Tissues were collected from ovariectomized ewes (OVX, n = 4), OVX-estradiol treated ewes (OVX-E, n = 5), and ewes (n = 7) slaughtered 5 d after parturition (PP). Following euthanasia and exsanguination, stalk-median eminence (SME), medial-basal hypothalamus (MBH) and preoptic areas (POA) were collected. Concentrations of GnRH and proGnRH were determined by radioimmunoassay using specific antisera. Concentrations of GnRH in the SME did not differ (P > .05) between OVX-E and OVX ewes, but both groups contained less (P < .05) GnRH than the SME from PP ewes (4.4 +/- 0.7, 12.1 +/- 3.8 vs 24.3 +/- 5.1 fmol/mg tissue, respectively). Concentrations of proGnRH in SME mimicked those of GnRH and were less (P < .05) in OVX-E ewes than PP ewes, but were not different (P > .05) from those in OVX ewes (.34 +/- .34 vs 3.76 +/- 1.53 and 1.7 +/- .78 fmol/mg, respectively). In the MBH, OVX-E ewes had greater (P < .05) concentrations of GnRH than PP ewes (0.76 +/- 0.29 vs 0.24 +/- 0.04 fmol/mg) and OVX ewes were intermediate (0.41 +/- 0.13 fmol/mg).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Differential regulation of gonadotropin synthesis and release in ovariectomized ewes after treatment with a luteinizing hormone-releasing hormone antagonist.

Our working hypothesis was that synthesis and release of LH, but not FSH, were solely dependent on LHRH. Twenty ovariectomized (OVX) ewes were randomly assigned to one of five treatments (n = 4 per group). Ewes were administered a low (10 micrograms/kg) or high (100 micrograms/kg) dose of LHRH antagonist (LHRH-Ant) at 24-h intervals for 3 or 6 days. Control ewes received vehicle (5% mannitol) at 24-h intervals for 6 days. Blood samples were collected every 15 min for 4 h before LHRH-Ant or vehicle and every 2 h during the period of treatment to determine concentrations of LH and FSH. Twenty-four hours after the last treatment with LHRH-Ant or vehicle, anterior pituitaries were collected and divided in half along the midsagittal plane; the number of receptors for LHRH, pituitary content of LH and FSH, and relative amounts of mRNA for alpha, LH beta, and FSH beta subunits were determined. Concentrations of LH in serum decreased (p < 0.05) from 25.4 +/- 4.3 ng/ml before LHRH-Ant to less than 0.5 ng/ml within 4 h after the first treatment of LHRH-Ant and remained low (< 0.5 ng/ml) throughout the study. Serum concentrations of FSH declined gradually during the 3- or 6-day period of treatment with LHRH-Ant, from 37.3 +/- 2.4 and 26.5 +/- 4.8 ng/ml to 19.9 +/- 1.8 and 13.7 +/- 2.1 ng/ml, respectively. The magnitude of decline in serum concentrations of LH and FSH did not differ among ewes treated with low or high doses of LHRH-Ant.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Immunization of mice against a synthetic N-terminal extracellular domain gonadotropin-releasing hormone receptor peptide: evidence for a direct uterine effect.

PROBLEM: Immature male and female mice were immunized with a synthetic peptide corresponding to amino acids 5-17 (ASLEQDPNHCSAI) of the mouse hypophyseal gonadotropin-releasing hormone (GnRH) receptor. METHOD: Effect of immunization (postpuberal) was restricted to the uterus. Pituitary-gonadal functions were not altered. RESULTS: The endometrial lining of immunized females was thin and lacked glandular development. These observations were corroborated in actively immunized and passively immunized adult females. CONCLUSIONS: Apparently endometrial cells express a unique surface antigen, though reactive with antipeptide antibodies, that differs from the prototype pituitary GnRH receptor. Antibodies that selectively inhibit endometrial maturation might be used to treat proliferative diseases of the uterus.

Amino Acid Sequence↗

PubMed1993

Hypothalamic and hypophyseal receptors for estradiol in high and low sexually performing rams.

Rams, characterized over a 3-year period as being high (n = 8) or low (n = 10) sexual performers, were euthanized to determine if differences in sexual behavior were associated with differences in hypophyseal and hypothalamic receptors for estradiol. Rams were exposed to estrous ewes and courtship behavior was recorded for 5 hr prior to tissue collection. Blood samples for determination of serum concentrations of luteinizing hormone (LH) were collected every 15 min during the behavior testing period. Hourly blood samples were analyzed for serum concentrations of estradiol (E) and testosterone (T). Rams characterized as high (HP) or low (LP) sexual performers had similar LH profiles; there were no differences noted in basal concentration, number of pulses, or pulse amplitude of LH. There were no differences in serum concentrations of T or E during the behavior testing period between HP and LP rams. Unoccupied receptors for E (moles E bound x 10(-15)) in the stalk-median eminence (SME) (P < 0.05) in HP rams (15.1 +/- 3.1) were greater than those in LP rams (4.6 +/- 1.9). Occupied receptors for E (moles E bound x 10(-15)) in the preoptic area (POA) (P < 0.1) in HP rams (6.0 +/- 0.8) tended to be greater than those in the LP rams (3.8 +/- 0.8). The proportion of total E receptors (occupied+unoccupied) in the POA that were occupied was greater (P < 0.05; 0.41 +/- 0.03 vs 0.23 +/- 0.04) for HP than for LP rams. In contrast, HP rams had less (P < 0.05) occupied E receptors (moles E bound x 10(-15)) in the anterior pituitary gland (AP) than LP rams (152.0 +/- 38.5 vs 337.3 +/- 51.7). The proportion (%) of E receptors that were occupied in the AP in HP rams also tended to be less (P < 0.06) than that in LP rams (8 +/- 2 vs 14 +/- 1). No differences were detected in occupied or unoccupied E receptors in the medial basal hypothalamus, medial amygdala, or stalk-median eminence. In conclusion, differences in occupied receptors for E in the POA and the AP, and unoccupied receptors for E in the SME, were associated with differences in ram sexual behavior.

Amygdala↗

PubMed1993

Effects of season and estradiol on concentrations of histamine within discrete brain regions of ovariectomized ewes.

Histamine has been implicated as a neuromodulator of secretion of gonadotropins in several species. Concentrations of histamine were analyzed within discrete brain regions and endocrine tissues to help determine whether this amine has the potential to exert a similar function in ewes expected to have dramatically different serum concentrations of LH. Following collection of blood samples at 12-min intervals for 4-hr, ovariectomized (OVX) and OVX-estradiol treated (OVX-E) ewes were slaughtered during the breeding and anestrous seasons (five animals/group). Concentrations of LH were depressed by treatment with estradiol (E; P < .01), but to a greater extent (P < .05) during the anestrous season compared to the breeding season. Concentrations of histamine in tissues (ng/mg) differed (P < .01) between the breeding and anestrous seasons, in the medial thalamus (39.2 +/- 14.1 vs 109.9 +/- 13.0), posterior pituitary gland (247.6 +/- 50.7 vs 23.0 +/- 9.1) and midbrain tegmentum (10.4 +/- 5.6 vs 50.7 +/- 3.9). Estradiol containing implants decreased (P < .05) concentrations of histamine in the midbrain tegmentum (20.3 +/- 7.1 vs 37.7 +/- 7.8) and posterior pituitary gland (87.3 +/- 24.0 vs 258.2 +2- 67.5) compared to non-estradiol treated controls. Histamine concentrations in the pineal and anterior pituitary glands and brain regions; stalk-median eminence, medial basal hypothalamus, preoptic area, cerebellum, parietal neocortex, were not (P > .05) affected solely by either season or E. An interaction between effects of season and estradiol on concentrations of histamine occurred (P < .05) in the posterior pituitary gland and the preoptic area.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Relationship of body composition of mature ewes with condition score and body weight.

Fourteen mature, ovariectomized, western-range ewes with an initial mean BW of 72 +/- 4.5 kg and mean condition score (CS) of 7.5 +/- .3 were used to evaluate the relationship between CS and body composition. Diets of chopped straw and alfalfa hay were formulated to provide either maintenance energy or less than maintenance energy (100 or 60% of ME) to induce changes in BW and CS. After 180 d, ewes were weighted, scored for body condition, and slaughtered. All carcass components, viscera, and organs were analyzed for lipid, DM, and ash, and protein was determined by difference. Body weight and CS values were related by regression analysis to percentage of composition and weights of carcass components, carcass, and empty body. Body weight and CS were highly correlated (r = .89) and analysis indicated that each increase in CS resulted in an increase of 5.1 kg of BW. Condition score accounted for more variation of percentage of lipid in the empty body (R2 = .95) and carcass (R2 = .90) than did BW (R2 = .84 and .80, respectively). In contrast, BW accounted for more of the variation in carcass weight (R2 = .97) and empty BW (R2 = .99). Inclusion of both BW and CS in regression models did not increase the variation accounted for with the single best predictor. With mature western-range ewes in this study, CS was highly related to carcass lipids and could be used to describe energy reserves available to ewes.

Adipose Tissue↗

PubMed1992

Effect of implantation of dexamethasone adjacent to the paraventricular nucleus on messenger ribonucleic acid for corticotropin-releasing hormone and proopiomelanocortin during late gestation in fetal sheep.

Glucocorticoids act upon the hypothalamic paraventricular nucleus (PVN) and anterior pituitary in a classic negative feedback loop to regulate ACTH biosynthesis and secretion. Evidence exists to indicate that glucocorticoid feedback may be attenuated during late gestation in the sheep fetus to allow the preterm rise in fetal plasma cortisol necessary for parturition in this species. The present studies were undertaken to determine the effect of glucocorticoids placed adjacent to the fetal PVN on messenger RNA (mRNA) for CRH in the PVN and mRNA for POMC in the anterior pituitary during late gestation. We performed our studies at two critical stages during late gestation to determine if gestational age related changes occur in the efficacy of negative feedback regulation of expression of CRH and subsequently POMC. Dexamethasone (DEX) implants were placed bilaterally 2 mm lateral to the fetal PVN at 105 to 107 days gestational age (dGA; group I, n = 4) and 121-123 dGA (group II; n = 4). Gestational-age matched, sham implanted fetuses were used as controls (CONT) for both groups (n = 4 per group). Fetuses were recovered at 126-128 (group I) and 136 dGA (group II). Fetal PVN were isolated by micropunching, and the anterior pituitary was separated from neurointermediate and posterior lobes after necropsy. Total RNA was subjected to Northern analysis using specific complementary DNA probes to CRH and POMC, and specific message was normalized to actin mRNA content in each individual sample. Anterior pituitary POMC mRNA was not different in DEX fetuses compared to CONT for either group I (78 +/- 26% of CONT; mean +/- SEM) or group II (84 +/- 17% of CONT). PVN CRH mRNA content was lower in DEX fetuses in group I (28 +/- 14% of CONT; P less than or equal to 0.01) and group II (65 +/- 12% of CONT; P less than or equal to 0.01). The degree to which DEX suppressed mRNA for CRH was greater in group I compared to group II (P less than or equal to 0.05). We conclude that 1) CRH expression in the PVN of fetal sheep is suppressible by glucocorticoids; 2) suppression can occur directly at the level of the PVN and 3) that the efficacy of negative feedback decreases with increasing gestational age. Furthermore, the lack of effect of hypothalamic administration of DEX on anterior pituitary POMC mRNA indicates that basal expression of POMC in fetal sheep may be independent from support from the PVN at this stage of gestation.

Animals↗