Effects of leptin on gonadotropin-releasing hormone release from hypothalamic-infundibular explants and gonadotropin release from adenohypophyseal primary cell cultures: further evidence that fully nourished cattle are resistant to leptin.

In rodents and pigs, leptin stimulates the release of gonadotropin-releasing hormone (GnRH) from hypothalamus, gonadotropins from adenohypophyseal (AP) explants and cells, and luteinizing hormone (LH) from full-fed animals. In the current studies, we investigated whether leptin could stimulate the release of GnRH from bovine hypothalamic-infundibular (HYP) explants and gonadotropins from bovine adenohypophyseal cells. In Experiment 1A, HYP explants collected from 17 bulls and seven steers were incubated with Krebs-Ringer bicarbonate buffer (KRB) containing 0, 10, 100, or 1000 ng/ml recombinant ovine leptin (oleptin) for 30 min after a 3-h period of equilibration. None of the doses of leptin affected (P > 0.05) GnRH release into the media. In Experiment 1B, HYP explants collected from six steers were incubated with KRB containing 0 or 1000 ng/ml oleptin for two consecutive 30-min periods and challenged with 60 mM K(+) afterwards. Leptin did not affect (P > 0.05) basal or K(+)-stimulated release of GnRH. In Experiment 2, adenohypophyses from steers were collected at slaughter and cells dispersed and cultured for 4 days. On day 5, cells were treated with media alone (control) or media containing 10(-11), 10(-10), 10(-9), and 10(-8)M oleptin. Three independent replications were performed. None of the doses of leptin stimulated (P > 0.05) the release of LH. Although leptin at 10(-11), 10(-10), and 10(-9)M increased (P < 0.03) slightly the release of FSH compared to control-treated cells in one replicate, this effect was not confirmed in the other two replicates. Results support the hypothesis that leptin has limited effects on the release of GnRH and gonadotropins in full-fed cattle and reiterate important species differences in responsiveness to leptin.

Effects of leptin on basal and GHRH-stimulated GH secretion from the bovine adenohypophysis are dependent upon nutritional status.

We have shown recently that leptin modulates at least two aspects of anterior pituitary LH release in ruminants: basal and GnRH-mediated release. To test the hypothesis that leptin directly affects basal and GHRH-mediated GH secretion from the adenohypophysis, we examined the effects of various doses of recombinant ovine leptin (oleptin) on perifused adenohypophyseal (AP) explants and compared responses of tIssues from control and fasted cows. Ten mature, ovariectomized and estradiol-implanted cows were assigned to one of two dietary groups: (1) normal-fed (n=5) and (2) fasted for 72 h (n=5). At the end of the fasting period, cows were euthanized and pituitaries were collected. Adenohypophyseal explants were perifused for a total of 6.5 h, including a 2-h treatment at 2.5 h with Krebs-Ringer bicarbonate buffer containing 0, 5, 10, 50, or 100 ng/ml oleptin, and a challenge with GHRH at 4.5 h. All doses of oleptin greater than 5 ng/ml decreased (P<0.01) basal GH secretion compared with controls in tIssues collected from normal-fed cows. In contrast, GH release from AP explants from fasted cows treated with the lowest dose of oleptin was 28% (P<0.002) higher than control explants, but larger doses had no effect. Leptin caused an inversely related, dose-dependent increase in GHRH-mediated GH release in tIssues from normal-fed cows. Marked increases (P<0.01-P<0.001) in GH release were observed for the 5 and 10 ng/ml oleptin, with lesser (P<0.08) and no effects observed at the 50 and 100 ng/ml doses respectively. In fasted cows, oleptin had no stimulatory effect on GHRH-induced GH release. Results show that leptin can act directly at the anterior pituitary level to modulate GH release, and this effect is dependent upon nutritional history.

Leptin acts at the bovine adenohypophysis to enhance basal and gonadotropin-releasing hormone-mediated release of luteinizing hormone: differential effects are dependent upon nutritional history.

Recombinant ovine leptin (oleptin) stimulates an acute increase in the secretion of LH in fasted, but not in normal-fed, cows through an augmentation of the magnitude of individual pulses of LH. Herein, we tested the hypothesis that this effect could be accounted for by functional changes at the adenohypophyseal (AP) level. Eleven ovariectomized, estradiol-implanted cows were assigned to one of two dietary groups: normal-fed (n = 6) and fasted (fasted for 72 h; n = 5). After the animals were killed, the adenohypophyses were collected and AP explants were perifused with Krebs-Ringer bicarbonate buffer (KRB) for a total of 6.5 h, including a 2-h treatment at 2.5 h with KRB or increasing doses of oleptin and a challenge at 4.5 h with 50 ng of GnRH. To test for effects of leptin at the hypothalamic level, explants encompassing the medial basal hypothalamus-infundibular complex (HYP) were incubated in KRB alone (control) or in KRB containing 1000 ng of oleptin. Basal release of LH from AP explants treated with leptin was greater (P < 0.02) than that from control-treated explants in fasted, but not in normal-fed, cows. To the contrary, leptin-treated explants from normal-fed, but not from fasted, cows released more (P < 0.001) LH in response to GnRH than control-treated tissues. Neither fasting nor leptin affected (P > 0.1) the secretion of GnRH from HYP explants. These observations support the hypothesis that leptin modulates the secretion of LH in mature cows, to a large extent, by its direct actions at the AP. Differential manifestations of these effects are dependent upon nutritional history.

Calcium/calmodulin-dependent protein kinase II involvement in release of gonadotropin-releasing hormone.

Involvement of calcium/calmodulin-dependent protein kinase II (CaM kinase II) in regulation of GnRH release was tested by determining the effect of CaM kinase II antagonists (KN-62 or KN-93) on GnRH release from rat or cattle infundibular (stalk median eminence) explants. Preincubation of male rat infundibular explants for 30 min with KN-62 (0.5, 1, 5 or 10 microM) 1.5 h prior to the addition of 59.3 mM (high) K+ resulted in a dose-dependent suppression of GnRH release. A longer pretreatment period (2 h) of rat infundibular explants with KN-62 (1 or 10 microM) appeared to enhance the suppressive effect of the CaM kinase II antagonist. Exposure (2 h) of rat infundibular explants to 10 microM, but not 0.1 microM KN-93, resulted in a complete inhibition of high K+-induced GnRH release. Exposure of steer infundibular explant halves to KN-62 (50 or 100 microM) or KN-93 (50 microM) inhibited high K+-induced GnRH release. Likewise, treatment of heifer infundibular explant halves with KN-93 (50 microM) abolished high K+-induced GnRH release. The period of exposure required for KN-62 to elicit its effect was relatively short since exposure of KN-62 (100 microM) for only 91-150 min of incubation was sufficient to block high K+-induced GnRH release from steer infundibular explant halves. In conclusion, these results: (1) support the hypothesis that CaM kinase II is involved in GnRH release from the rat and cattle infundibulum, (2) demonstrate that the effect of CaM kinase II on GnRH release from cattle infundibula is independent of reproductive state, (3) confirm previous reports supporting Ca2+ and CaM involvement in GnRH release from rat and cattle infundibula and (4) establish that infundibular explants incubated in vitro are useful for studying selected mechanisms regulating hypothalamic neurohormone release from neuron terminals.

Effects of exogenous recombinant ovine interferon tau on circulating concentrations of progesterone, cortisol, luteinizing hormone, and antiviral activity; interestrous interval; rectal temperature; and uterine response to oxytocin in cyclic ewes.

Interferon tau (IFN tau) is the conceptus-produced antiluteolytic signal in ruminants. Three experiments examined the effects of s.c. administration of recombinant ovine (ro)IFN tau on interestrous interval (IEI), oxytocin (OT)-induced uterine prostaglandin F2alpha metabolite (PGFM) production, rectal temperature (RT), respiration rate (RR), and plasma concentrations of progesterone, cortisol, LH, and antiviral activity (AVA) in plasma and uterine flushings. In experiment I, 20 ewes were treated s.c. with either 0, 1, 2, or 4 mg/day roIFN tau (0.7 x 10(8) U/mg; 5 ewes/dosage) from Days 11 to 15 of the estrous cycle (estrus = Day 0) and were challenged with OT (30 IU) on Day 15. Jugular blood samples were collected at -10, 0, 10, 20, 30, 40, 50, and 60 min relative to the OT challenge and assayed for PGFM. Recombinant oIFN tau increased IEI (16.7, 18.7, and 22.6 +/- 0.6 days for 0, 2, and 4 mg roIFN tau, respectively, p < 0.01). Recombinant oIFN tau did not affect peak PGFM response to OT (2309 +/- 172 pg/ml; p > 0.1). However, the 4 mg/day dosage delayed the time to peak PGFM (32.4 vs. 47.5 +/- 3.4 min; p < 0.01, 0 vs. 4 mg) and resulted in approximately 200% higher concentrations of PGFM at 60 min post-OT (0 vs. 4 mg/day, p < 0.07). Experiment II was similar to experiment I, except that only the 0- and 4-mg/day dosages of roIFN tau were administered. Ewes were hysterectomized on Day 16, and assay of uterine flushes detected no AVA from ewes treated with either 0 or 4 mg/day roIFN tau. In experiment III, 20 ewes were treated s.c. with either 0, 2, 4, or 6 mg roIFN tau on Day 12. Blood samples, RT, and RR were obtained at frequent intervals for 24 h, and plasma was assayed for progesterone, cortisol, LH, and AVA. Plasma AVA, which increased in a dose-dependent manner, was detectable within 60 min and remained elevated at 24 h compared to control values. RT (elevated 0.5-1.0 degrees C), RR, and cortisol increased in response to all dosages of roIFN tau, with peak values occurring 150-180 min postinjection. For all dosages of roIFN tau, plasma progesterone declined from 120 to 360 min posttreatment and then returned to pretreatment values by 24 h (p < 0.01) as compared to controls. Overall, exogenous roIFN tau altered uterine PGFM response to OT from a pulse to a gradual and sustained elevation and extended IEI with only a transient decline in progesterone and mild hyperthermia, effects that are not expected to compromise pregnancy.

Temporal aspects of ovarian follicular growth and steroidogenesis following exogenous follicle-stimulating hormone in Angus heifers.

Ultrasonography and endocrine assay techniques were used to monitor structural and hormonal alterations made by the ovary in response to the biological actions of pituitary-derived follicle-stimulating hormone (FSH-P). Angus heifers (n = 36) were allotted to receive injections (twice per day) of either FSH-P (up to a total of 28 mg over a maximum of 4 days beginning on Day 10 of a synchronized estrous cycle) or saline in order to quantify temporal relationships among follicle growth and steroid hormone profiles. Transrectal ultrasonography was utilized at 12-h intervals to monitor and record follicle growth. Plasma was collected every 12 h for the first 48 h of the experiment and then every 6 h for the remainder of the experiment. At 48 and 60 h after the onset of treatments, prostaglandin F2 alpha (PGF2 alpha; 25 mg) was administered (i.m.). FSH-treated heifers (n = 6 at each time) were terminated at 24, 48, 72 and 96 h following the onset of treatment. Saline-treated heifers were terminated at 24 and 96 h (n = 6 at each time). After ovaries were obtained, follicular number and size were recorded and follicular fluid (FF) was collected. Plasma concentration of progesterone (P) and estradiol (E2) and FF concentration of P, E2, estrone, testosterone and androstenedione were determined by radioimmunoassays. Plasma concentration of E2 increased (P < 0.05) within 36 h of initiation of FSH treatment. Plasma P decreased (P < 0.0001) by 12 h post-PGF2 alpha. Ultrasonographic examination revealed a significant decrease in the number of small follicles by 48 h, whereas the number of medium follicles increased (P < 0.05) by 60 h after the initiation of FSH treatment. The number of large follicles (LF > or = 10 mm diameter) increased (P < 0.01) over the course of the experiment. The total number of ovarian follicles (TF) 24 h after the start of FSH treatment was correlated (r = 0.99; P < 0.0001) with the number of small follicles (SF < or = 5 mm). At 72 h after the onset of FSH treatment, the number of medium follicles (i.e. 6-9 mm) was correlated with TF (r = 0.97; P < 0.0001). Estradiol was the predominant FF steroid. Follicular fluid E2 was greatest in follicles at 72 h after FSH treatment. Follicular fluid E2 and plasma E2 were positively correlated (r = 0.66; P < 0.001). Follicular aromatase activity was estimated by evaluating the ratio of FF estrogens (E) to androgens (A). Elevated aromatase activity (E:A ratio > 1.0) was detected in 196 of 206 follicles. The estrogen to progesterone ratio was used as an estimate of follicle viability. Eighty-five percent of the follicles were estimated to be viable (E:P ratio > 1.0). The peak E:A ratio in LF preceded by 24 h the peak concentration in FF E2 and plasma E2. In MF and SF the E:A ratio increased by 72 h. Enhancement of ovarian follicular growth (i.e. increased number and size of follicles; increased steroidogenesis) by exogenous, pituitary-derived FSH is characterized by (1) increased activity of aromatase, and (2) accumulation of FF E2, events which temporally preceded the increase in plasma concentration of E2. These observations will aid efforts to incorporate recombinant bovine FSH and somatotropin in an effort to develop more predictable superstimulation and ovulation induction protocols.

Effect of histone-H2A (H-H2A), platelet activating factor (PAF) and pregnancy specific protein B (PSPB) on secretion of prostaglandins E and F2 alpha (PGE; PGF2 alpha) by bovine endometrium and H-H2A on basal secretion of luteinizing hormone (LH) by bovine pituitary cells in vitro.

Effects of H-H2A, PSPB or PAF on day 16 bovine endometrial secretion of PGE and PGF2 alpha and H-H2A on basal secretion of LH by bovine pituitary cells in vitro were examined in two experiments. PAF (P < or = 0.08) and H-H2a + PAF (P < or = 0.10) treatment for two hours in an in vitro perfusion system tended to increase secretion of PGF2 alpha expressed as a proportion of the prechallenge concentrations of PGF2 alpha by day 16 bovine caruncular endometrium, which occurred during the two-hour period after treatment removal. PGF2 alpha was increased (P < or = 0.02) in the two hour period after the treatment was removed in both control and H-H2A-treated endometrium. H-H2A (P < or = 0.07) and PSPB (P < or = 0.08) tended to increase PGE, while H-H2A + PSPB (P < or = 0.05) and H-H2A + PAF (P < or = 0.03) increased secretion of PGE expressed as a proportion of the prechallenge concentrations of PGE by day 16 bovine caruncular endometrium during the challenge and postchallenge treatment periods, but did not differ (P < or = 0.05) between the two-hour challenge period and the two-hour postchallenge period after removal of the treatment. In experiment two, H-H2A decreased (P < or = 0.05) basal secretion of LH by bovine pituitary cells in vitro. These data suggest that H-H2A and PSPB preferentially stimulate secretion of PGE by bovine endometrial tissue and may play a role in maternal recognition of pregnancy, while PAF increased PGF2 alpha secretion. In addition, H-H2A may play a role in regulating secretion of LH by the pituitary. Key words: Pregnancy Specific Protein B, Histone-H2A, Prostaglandin E and F2 alpha, Platelet Activating Factor, Endometrium, Cow.

Calcium involvement in luteinizing hormone-releasing hormone release from the bovine infundibulum.

Bovine infundibular (stalk median eminence) explants were incubated in vitro to test the hypothesis that calcium (Ca) is involved in the release of luteinizing hormone-releasing hormone (LHRH) from LHRH neuron terminals in cattle. Right and left infundibular halves from individual heifers and/or steers were randomly assigned to either control or treated (EGTA [a Ca chelator] or verapamil [an L-type Ca channel antagonist]) groups. Each half was incubated in 600 microliters of Krebs-Ringer bicarbonate medium (KRB) in the presence or absence of a treatment agent for 180 min. At 30-min intervals, 500-microliters samples were removed from each incubated and replaced with fresh media. Spontaneous (basal) and depolarization-induced (60 mM potassium) LHRH release was evaluated by radioimmunoassay of the LHRH content in the media incubated from 91 to 120 and 121 to 150 min of culture, respectively. The effect of treatment on depolarization-induced LHRH release was analyzed by comparing the differences between spontaneous and depolarization-induced LHRH release in control and treated groups. Spontaneous LHRH release was not different between control and 1.25 mM EGTA- or 100 microM verapamil-treated halves from steers. In contrast, steer infundibular halves incubated with EGTA (replacing Ca in KRB and chelating any Ca in the media) released less LHRH during depolarization than did control halves. In addition, verapamil-treated (to block Ca uptake by the terminal) infundibular halves from steers or heifers released less LHRH in response to depolarization than did control halves.(ABSTRACT TRUNCATED AT 250 WORDS)

Regulation of adrenocorticotropin secretion in vitro by anterior pituitary corticotrophs from fallow deer (Dama dama).

The actions of corticotropin-releasing hormone (CRH), vasopressin (VP), the synthetic glucocorticoid dexamethasone (DEX), and mifepristone (RU 486), a glucocorticoid antagonist, on the secretion of adrenocorticotropin (ACTH) by cultured fallow deer corticotrophs were studied in vitro. On Day 5 of primary culture, corticotrophs were challenged for up to 4 hr with medium alone (Control), CRH, VP, DEX, forskolin (FSK), phorbol ester (TPA), cyclic AMP (cAMP), and/or RU 486 at various concentrations and combinations. CRH, VP, FSK and TPA each stimulated (P < 0.01) the secretion of ACTH in dose- and time-related manners. Relative to Control, CRH at 0.001 and 0.1 microM and VP at 0.01 and 1 microM increased (P < 0.01) medium concentration of ACTH by 7.3-, 13.5-, 3.7- and 9.0-fold, respectively. There was a treatment x incubation time interaction (P < 0.01) such that at 30-min posttreatment, CRH-induced ACTH secretion tended (P < 0.10) to be less than that obtained via VP treatment, whereas at 1, 3, and 4 hr posttreatment, medium concentration of ACTH from cells treated with 0.1 microM CRH was greater (P < 0.05) than that in cells treated with 1 microM VP. At equimolar doses of 0.01 and 0.1 microM, CRH was 3.4- and 3.0-fold more potent (treatment x dose, P < 0.05) than VP. Cotreatment with 1 microM DEX reduced (P < 0.001) the stimulatory effects of CRH (0.1 microM), VP (1 microM), FSK (10 microMs), TPA (0.1 microM), and cAMP (0.001 M). However, the coaddition of RU 486 (1 microM) to the CRH plus DEX- and the FSK plus DEX-treated wells partially negated the inhibitory effects of DEX. RU 486 completely negated the inhibitory effects of DEX on the VP-, TPA-, and cAMP-stimulated secretion of ACTH. These data indicate that CRH is a more potent stimulator of ACTH secretion than is VP in primary culture of fallow deer pituitary cells. This study also demonstrates the utility of an in vitro culture system to investigate stress-related hormonal interactions in cervids.

Luteinizing hormone-releasing hormone gene expression in the bovine brain: anatomical localization and regulation by ovarian state.

Experiments were conducted to identify neurons in the bovine brain that express the LHRH gene and to determine whether LHRH mRNA levels are influenced by the ovaries. Two groups of postpubertal heifers were utilized: heifers killed during the mid-luteal phase of the estrous cycle (LUTEAL, n = 5) and heifers killed 14-16 wk following ovariectomy (OVX, n = 5). In situ hybridization was performed through use of a 32P-end-labeled deoxyoligonucleotide (59 mer) complementary to the human LHRH mRNA sequence. LHRH-expressing neurons were identified in the diagonal band of Broca, the preoptic area, and the anterior hypothalamus in a manner consistent with immunocytochemical localization. Reduced silver grains, proportional to LHRH mRNA content, were quantified (in pixels, 45x objective) with an image analysis system. Expected serum hormone concentration differences between endocrine states were confirmed by radioimmunoassay for progesterone (LUTEAL > OVX, p < 0.01) and for LH (OVX > LUTEAL, p < 0.01). Compared to the OVX group, LUTEAL heifers had 34% fewer LHRH-expressing neurons (p < 0.05); on the average, these neurons possessed 28% fewer pixels/cell (p < 0.01), indicating fewer copies of LHRH mRNA per cell. When the numbers of pixels in all labeled cells were totalled, LUTEAL animals had 57% fewer pixels (p < 0.05) than did the OVX females--probably reflecting a decrease in LHRH synthetic capacity in the LUTEAL animals. Therefore, during the mid-luteal phase of the bovine estrous cycle, ovarian steroid (i.e., luteal progesterone) suppression of LHRH release (as reflected by serum LH) is coincident with decreased LHRH mRNA in the brain.(ABSTRACT TRUNCATED AT 250 WORDS)

A possible dual mechanism involved in the inhibitory influence of suckling on luteinizing hormone release in the ovariectomized rat.

A study was conducted to further understand involvement of the endogenous opioid peptides in suckling-induced inhibition of LH release in ovariectomized rats. The first experiment was designed to determine the effect of an opioid antagonist, naloxone (NAL, 1.0 mg. kg-1h-1), on the increase in peripheral LH concentration 18 h after pup removal and on the decrease in LH concentration 18 h after pup return. Infusion of NAL during the 18 h after pup removal or during the 18 h after pup return neither accentuated nor attenuated serum LH concentrations. The second experiment was designed to determine the effect of NAL on peripheral LH concentrations in continuously suckled rats. Serum LH increased (p less than 0.10 and p less than 0.005, respectively) in response to 18 and 36 h of NAL infusion. The third experiment was designed to determine the effect of pup removal during NAL infusion on serum LH. Peripheral LH concentrations were not different in the rats treated with 36 h of NAL infusion whether they were suckled for the duration of the infusion or nonsuckled for the last 18 h of infusion. These results suggest that suckling may inhibit LH release through two mechanisms. The first may be an opioid-independent or enhanced opioid tone mechanism important for the initiation of the inhibitory effect of suckling on LH release, while the second may be an opioid-dependent mechanism important for the sustained inhibitory effect of suckling on LH release.

GnRH in the infundibular stalk-median eminence is related to percentage body fat in carcasses of beef cows.

Mature Hereford cows (n = 28) were used to determine the effect of percentage body fat on secretion of LH and content of GnRH in the infundibular stalk-median eminence (ISME). Cows were fed to maintain, lose, or gain weight to achieve body condition scores (BCS; 1 = emaciated; 9 = obese) of 3 to 7. Then cows were fed to maintain weight and body condition. Before slaughter, estrus was synchronized using two injections of prostaglandin F2 alpha(PGF) 11 d apart. Five d after the second PGF injection, cows were given 100 micrograms of GnRH (im) and serum samples were obtained. LH was quantified using RIA. The anterior pituitary and ISME were obtained within 45 min of death. Anterior pituitary weight and LH concentration, total GnRH in the ISME, total carcass fat, and percentage carcass fat were determined. BCS of cows at the time of slaughter influenced percentage carcass fat (P less than .001), total GnRH in the ISME (P less than .02), and maximum LH after GnRH treatment (P less than .09), but did not influence pituitary weight or concentration of LH in the pituitary. Content of GnRH in the ISME averaged 76 +/- 12, 32 +/- 14, 27 +/- 13, and 24 +/- 13 ng for cows with BCS of 3, 5, 6, and 7, respectively. BCS was correlated (P less than .001) with percentage carcass fat (r = .94) and total fat in the carcass (r = .92). Total GnRH in the ISME was negatively correlated (P less than .005) with BCS (r = -.54), percentage carcass fat (r = -.55), and total carcass fat (r = -.49).(ABSTRACT TRUNCATED AT 250 WORDS)

A priming effect of naloxone on in vitro LHRH release from the hypothalamus of mid-luteal ewes.

The possible involvement of endogenous opioid peptides (EOPs) in LHRH release from hypothalami of ewes during the breeding season was investigated using an in vitro perifusion system. Hypothalami were procured in December from ovariectomized (OVX; 62-65 days before the experiment; n = 6) and mid-luteal (ML; n = 7) Western White-Face ewes. Hypothalami were mid-sagitally sectioned into halves containing the preoptic area, mediobasal hypothalamus, and infundibulum (median eminence). The left half (treated) received two 30-min challenges (beginning at 130 and 250 min, respectively, after onset of perifusion) of 500 microM naloxone (NAL) followed by a 30-min 60-mM potassium (K) challenge (at 370 min after onset of perifusion). The right half served as the control, receiving only K at the same time as the treated tissue. Both NAL challenges elicited (p less than 0.05) LHRH release from tissues of both ML and OVX ewes. Release of LHRH by hypothalami from ML, but not from OVX, ewes was greater (p less than 0.01) after the second than after the first NAL challenge. These results are consistent with the view that an inhibitory opioid influence exists on LHRH release from ovine hypothalami. The release of LHRH in response to NAL was dependent on the ovarian status in vivo since the priming effect of NAL on subsequent NAL-induced LHRH release occurred only from the hypothalami of ML ewes. We suggest from these results that EOPs may modulate LHRH release from ovine hypothalami in an ovarian steroid-dependent and independent manner.

Influence of dietary energy intake on prepubertal development of Brahman bulls.

Twelve Brahman bulls (paired by sire, weight and age) were assigned randomly and limit fed to gain either .10 to .25 (moderate gain; MG) or .75 to 1.0 (high gain: HG) kg.hd-1.d-1 to examine the effect of dietary energy on onset of puberty. Hip height (HH), scrotal circumference (SC) and serum samples (20 min for 6 h) were obtained at four times (AGE): 0, 56 and 112 d on feed and after appearance of first motile spermatozoa (FS) in the ejaculate of HG bull of the pair. At FS both bulls of a pair were slaughtered, reproductive tissues were collected and in vitro GnRH release from the median eminence (ME) was measured. Increases in BW, HH and SC were greater (P less than .05) in HG bulls. Basal ME GnRH secretion was greater (P less than .05) in HG bulls. Serum LH concentrations were unchanged by energy level (P greater than .10) but increased (P less than .01) with increasing AGE. AGE and energy level increased (P less than .01) basal, mean and total serum testosterone (T) and these two factors acted synergistically (P less than .01). Height and amplitude of T pulses were increased by energy level (P less than .003) and AGE (P less than .002). Testicular T (P less than .08) and development (P less than .05) were increased in HG bulls. Growth hormone peak height and amplitude concentrations following feeding increased with AGE (P less than .06) but were not altered (P greater than .10) by energy level. Serum triglycerides (P less than .03) and BUN (P less than .003) increased with increasing AGE (P greater than .01). These data indicate that dietary energy level influences onset of puberty most directly at the testicular level.

Suckling inhibits release of luteinizing hormone-releasing hormone from the bovine median eminence following ovariectomy.

The effect of suckling on depletion of hypothalamic LHRH from the median eminence (ME) following ovariectomy (OVX) was determined in cattle. Multiparous, postpartum Holstein cows were assigned randomly to three groups: intact, nonsuckled (INT, n = 4); ovariectomized (3 to 5 d after parturition), nonsuckled (OVX, n = 4); and ovariectomized (3 to 5 d after parturition) and suckled by three calves (OVX-S, n = 5). Blood samples were collected at three periods (1 to 7 d before parturition and 3 to 5 d and 31 to 37 d after parturition) to determine plasma LH concentration. At 31 to 37 d after parturition, all cows were slaughtered and each ME was collected and mid-sagitally sectioned. The left half of each ME was used to determine content and concentration of LHRH. Concentrations of LH and LHRH were determined by RIA. Plasma LH concentration was similar among the three groups at 1 to 7 d before parturition and 3 to 5 d after parturition; however, at 31 to 37 d after parturition, OVX cows had a greater (P less than .05) concentration of LH (2.25 +/- .64 ng/ml) than either INT (.47 +/- .10 ng/ml) or OVX-S (.92 +/- .14 ng/ml) cows. Content of LHRH in the ME of INT (80.12 +/- 15.0 ng) and OVX-S 109.8 +/- 16.4 ng) cows was similar but was greater (P less than .05) than that in OVX cows (48.95 +/- 5.9 ng).(ABSTRACT TRUNCATED AT 250 WORDS)

Postpartum reproductive performance in the multiparous mare fed to obesity.

Twenty multiparous Quarter Horse mares were assigned to one of two treatment groups at 40 to 75 d of pregnancy. Group 1 was the control group and the mares were fed to maintain a moderate degree of body fat (condition score 5.5 to 7). Group 2 was the obese group and the mares were fed to achieve (prepartum) and then maintain (post partum) an extremely high degree of body fat (condition score 9). Estrous intensity was evaluated using subjective teasing scores ranging from 0 (rejection) to 4 (maximum receptivity). Mares were artificially inseminated beginning with the second postpartum ovulatory cycle; the study was terminated after 63 d of pregnancy. Duration of estrus, maximum teasing score and the number of mares exhibiting overt estrus (teasing score > 2) did not differ between treatment groups during the first and second postpartum ovulatory cycles. The intervals from foaling to first cycle ovulation, foaling to second cycle ovulation, and first to second cycle ovulation were also similar between treatment groups. All mares in both treatment groups conceived and maintained pregnancy. The first cycle conception rate and the number of cycles per conception did not differ between treatment groups. A high degree of body fat produced by overfeeding during gestation did not adversely affect postpartum reproductive performance in the multiparous mare.

Ovarian inhibition of peripheral plasma concentration of follicle stimulating hormone in prepuberal Holstein heifers.

The objective of this study was to determine effects of age and castration on follicle stimulating hormone (FSH) secretion in prepuberal heifers. In experiment 1, twelve heifers were bilaterally ovariectomized at 3, 6, or 9 months of age (n = 4/group). Blood was collected at 10 min intervals for 8 hr at 1 week before ovariectomy and 1 and 4 weeks after ovariectomy. Frequency, amplitude and duration of FSH pulses were calculated. Mean plasma concentration of FSH (ng/ml), and frequency (pulses/8 hr), amplitude (ng/ml), and duration (min/pulse) of FSH pulses were not altered by age. Mean concentration of FSH increased (P less than .01) from 1 week before to 1 week and 4 weeks after ovariectomy, respectively, in all age groups. Pulse frequency increased (P less than .05) from 1 week before ovariectomy to 4 weeks after ovariectomy in 3 month old heifers, from 1 week before to 4 weeks after ovariectomy in 6 month old heifers, and from 1 week before to 1 week and 4 weeks after ovariectomy in 9 month old heifers. In experiment 2, twelve heifers were bilaterally ovariectomized at 3, 6 or 9 weeks of age (n = 4/group). Sample collection and measurement of mean concentration of FSH were the same as in experiment 1. Mean concentration of FSH increased (P less than .01) from 1 week before to 1 and 4 weeks after ovariectomy in heifers ovariectomized at 6 and 9 weeks of age.(ABSTRACT TRUNCATED AT 250 WORDS)

Biological activity of LH during the peripartum period and the estrous cycle of the ewe.

Biological activity of luteinizing hormone (LH) is related to the degree of glycosylation of the glycoprotein hormone. The objectives of this study were to determine changes in biologically (BLH) and immunologically (ILH) active LH concentrations in plasma (in vitro bioassay and radioimmunoassay, respectively) and in the ratio of BLH to ILH (B:I) during the peripartum period and during the estrous cycle of the ewe. Blood samples were collected daily 4 days before through 4 days after parturition and during one estrous cycle. Also, samples were collected at 15-min intervals for 6 hr on Days 3 and 12 of the estrous cycle to quantify the influence of an elevated plasma concentration of progesterone (P) on the episodic secretion profiles of BLH. Progesterone concentration was determined on the 4th days pre- and post-partum, on each day of the estrous cycle and at hourly intervals on Days 3 and 12 of the cycle to investigate the hypothesis of an inverse relationship between P and BLH. The BLH and ILH concentrations were low during the peripartum period, and the B:I ratio did not increase by the 4th day postpartum. Mean ILH concentration was greater (P less than .05) in the postpartum than during the prepartum period. During the estrous cycle, mean daily B:I ratio was consistently above unity except for the day of estrus. The pre-ovulatory LH surge (BLH and ILH) was associated with a decrease (P less than .05) in the mean B:I ratio to 0.0065. Mean concentrations of BLH and ILH in plasma samples collected every 15 min on Day 12 were similar to Day 3 of the cycle.(ABSTRACT TRUNCATED AT 250 WORDS)

LHRH and LH in peripubertal female rats following prenatal and/or postnatal ethanol exposure.

The effects of pre- and postnatal exposure to ethanol (ETOH) on LHRH and LH were investigated. Pregnant and/or lactating dams were fed ETOH during: 1) gestation, 2) lactation, or 3) gestation-lactation. Female offspring were decapitated at 30 or 40 days-of-age; trunk blood was collected for plasma LH RIA; and hypothalamic tissues were collected for LHRH RIA. Hypothalamic LHRH content of all ETOH-exposed groups was less than that of non-ETOH-fed controls at 30 and 40 days-of-age (p less than 0.05). Plasma LH concentrations of all ETOH-exposed groups were less than those of non-ETOH-fed controls at 30 and 40 days-of-age (p less than 0.05). Also, at 30 and 40 days-of-age, the plasma LH concentrations of the animals exposed to ETOH during lactation and gestation-lactation were less than those of the animals exposed to ETOH during gestation (p less than 0.05). These data suggest that ETOH exposure during gestation and/or lactation negatively affects hypothalamic LHRH content of female rat offspring. Decreased hypothalamic LHRH content with corresponding lowered plasma LH concentration suggests that ETOH influences development or maturation of hypothalamic LHRH neurons by possibly decreasing their number or synthesizing capability.