Changes in luteinizing hormone secretion after estradiol treatment in prepubertal Nelore heifers

Changes in luteinizing hormone (LH) secretion after 17β-estradiol (E(2)) injection were evaluated during sexual maturation in 10 prepubertal Nelore heifers. Heifers were divided into 2 groups: intact (I) and ovariectomized (OVX). 17β-estradiol (2 μg/kg) was administered to both groups at 10, 13, and 17 mo of age. Only at 10 mo of age was there a greater mean LH concentration in OVX heifers (1.33 ± 0.29 ng/mL) compared with the I group (0.57 ± 0.15 ng/mL). At 13 and 17 mo of age there was no significant difference between the 2 groups in any of the evaluated variables (number of peaks, total peak area, greatest peak area, and time to greatest peak occurrence). This suggests a decrease in negative E(2) feedback associated with an increase in positive feedback to LH secretion during sexual maturation, and these were likely the key factors that determined the time of first ovulation in Nelore heifers.     

Negative feedback control of luteinizing hormone secretion in prepubertal beef heifers at 60 and 200 days of age

Prepubertal beef heifers at 60 and 200 d of age, born in the fall or spring, were assigned randomly to one of three treatment groups: (1) intact = 1; (2) bilateral ovariectomy (OVX); or (3) OVX plus estradiol-17 beta(E2) administered in silastic implants (OVX + E2). Luteinizing hormone (LH) was measured in serum samples collected at 20-min intervals for 4 h from heifers on -1, +7, +21, +35 and +49 d after OVX. Luteinizing hormone concentrations increased in the serum by 7 d after OVX in heifers at both 60 and 200 d of age (P less than .001; time X treatment). Prior to OVX, the LH patterns were characterized by low levels and infrequent episodic pulses. By 49 d after OVX, the mean LH concentrations increased and the pattern changed to one of rhythmic LH pulses with a periodicity of 1 h (P less than .001; time X treatment). Estradiol-treated OVX heifers did not exhibit a postovariectomy rise in serum LH concentrations. Serum E2 concentration 49 d after OVX in OVX heifers was threefold greater than in 1 or OVX heifers, thus demonstrating that E2 exerted negative feedback on pituitary LH secretion in prepubertal heifers. There was no measurable difference in serum E2 concentrations between I and OVX heifers; however, the contrast in the concentration and pattern of serum LH between the two groups was dramatic and suggested gonadal factors in addition to E2 are involved in controlling LH secretion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hypothalamic deafferentation in prepuberal beef heifers: Effects of gonadotropin-releasing hormone and estradiol benzoate on luteinizing hormone secretion

Hypothalamic control of luteinizing hormone (LH) secretion was investigated in crossbred beef heifer calves by comparing anterior (AHD), posterior (PHD), and complete (CHD) hypothalamic deafferentation with sham operated controls (SOC). Heifers (n = 16) were fitted with an indwelling jugular catheter for 6 days before cranial surgery, and assigned randomly to treatments. Blood for radioimmunoassay of LH was collected sequentially at 15-min intervals during an 8-h period on days ? 1 before and day 6 after hypothalamic deafferentation or sham operation. On the day of surgery, blood samples were collected sequentially at 15-min intervals 2 h before induction of anesthesia and throughout surgery and early recovery. Seven months after hypothalamic deafferentation, all experimental and sham operated heifers were ovariectomized and treated with vegetable oil (i.m.) plus saline (i.v.), vegetable oil plus gonadotropin releasing hormone (GnRH), estradiol benzoate (EB, 1 mg) in vegetable oil. After ovariectomy basal plasma concentrations of LH increased (P < 0.01) compared with the low circulating hormone levels before ovariectomy. The amplitude of LH response to GnRH was greater (P < 0.01) in CHD and PHD when compared with SOC and AHD heifers. Injection of EB failed to induce a LH surge in CHD and PHD 900–1100 min later when compared with the robust response seen in SOC and AHD heifers. Injection of EB plus GnRH elicited LH release in all deafferentated and sham operated heifers. These results indicate a transient change in LH secretion after AHD or CHD in prepuberal heifers with intact ovaries. After OVX, the integrity of the neural connection of the posterior hypothalamus is required for EB-induced LH release in beef heifers.     

Effects of restriction of dietary energy intake during the prepubertal period on secretion of luteinizing hormone and responsiveness of the pituitary to luteinizing hormone-releasing hormone in heifers

The working hypothesis that a low plane of nutrition during the prepubertal period delays puberty in heifers by retarding the prepubertal increase in secretion of luteinizing hormone (LH) was investigated. Secretion of LH and the responsiveness of the pituitary to LH-releasing hormone (LHRH) were compared in heifers fed a growing diet (which allowed spontaneous occurrence of puberty; n = 12; control) or an energy deficient diet (which delayed puberty; n = 11; delayed) during the prepubertal period. The dietary treatments were initiated when the heifers were 299 +/- 14 (mean +/- SD) d of age (d 0 of the experiment) and continued until d 175 of the experiment (474 +/- 14 d of age). Weight gains were .79 +/- .05 (mean +/- SE) and .21 +/- .03 kg X head-1 X d-1 for control and delayed heifers, respectively. Puberty occurred on d 120 +/- 14 of the experiment (428 +/- 13 d of age) in control heifers, whereas none of the delayed heifers attained puberty during the feeding period. Serum concentration of LH and the frequency of LH pulses increased rapidly during the 175-d feeding period in control heifers. In delayed heifers, serum LH concentration increased less rapidly and no increase in pulse frequency was detected during the experimental period. Amplitude of LH pulses tended to be higher in control than delayed heifers. Responsiveness of LH secretion to LHRH was lower in delayed than control heifers. It is speculated that failure of secretion of LH to increase is the causative factor for delayed puberty when dietary energy is limited during the prepubertal period in heifers.     

Induction of precocious puberty in heifers III: hastened reduction of estradiol negative feedback on secretion of luteinizing hormone

Precocious puberty (<300 d of age) can be induced in beef heifers by early weaning and feeding a high-concentrate diet. The objective of this experiment was to determine whether precocious puberty occurs as a result of a hastened reduction of estradiol negative feedback on secretion of LH. Thirty crossbred Angus and Simmental heifers were weaned at 83 +/- 2 d of age and 114 +/- 3 kg of BW, blocked by BW, and randomly assigned to receive a high-concentrate (60% corn; H) or control (30% corn; C) diet and to receive ovariectomy (OVX), OVX plus an estradiol implant (OVXE), or to remain intact (INT). Residual ovarian tissue after OVX necessitated withdrawal of 6 heifers during the course of the experiment, resulting in the following treatment groups: OVX-C, n = 3; OVX-H, n = 5; OVXE-C, n = 4; OVXE-H, n = 2; INT-C, n = 5; INT-H, n = 5. To determine concentrations of progesterone and estradiol, blood samples were collected weekly beginning at a mean age of 160 d. To characterize LH concentrations, serial blood samples were collected at 12-min intervals for 12 h at mean ages of 119, 149, 188, 217, 246, 281, 323, 365, 407, and 449 d. By a mean age of 202 d, heifers fed the H diet were heavier (P < 0.05) than those fed the C diet. Heifers in the INT-H treatment attained puberty earlier (P < 0.05) than in the INT-C treatment (275 +/- 30 vs. 385 +/- 14 d of age, respectively). Overall mean concentrations of estradiol did not differ between OVXE-H and OVXE-C, between INT-H and INT-C, or between OVXE and INT treatments. The OVX treatments exhibited greater LH pulse frequency than the OVXE and INT treatments by the first serial blood collection (treatment x age, P < 0.05). The frequency of LH pulses was greater (P < 0.05) in the INT-H than the INT-C treatment by a mean age of 246 d and was greater (P < 0.05) in the OVXE-H than the OVXE-C treatment by a mean age of 281 d. In the OVXE-H treatment, LH secretion increased and subsequently "escaped" from estradiol negative feedback (detection of > or = 1 LH pulse/h) earlier (P < 0.05) than in the OVXE-C treatment (307 +/- 30 and 420 +/- 21 d of age, respectively). It is concluded that advancing the reduction of estradiol negative feedback on secretion of LH is the mechanism by which early weaning and feeding a high-concentrate diet results in precocious puberty in heifers.     

Pulsatile or continuous infusion of luteinizing hormone-releasing hormone and hormonal concentrations in prepubertal beef heifers

An experiment was conducted to determine if exogenous luteinizing hormone-releasing hormone (LHRH) administered iv intermittently as pulses (P) or by continuous sc infusion (I) using osmotic minipumps could sustain pulsatile LH release and induce estrous cyclicity in prepubertal heifers. Prepubertal heifers were assigned randomly to: 1) receive pulses of LHRH (n = 6; 2.5 micrograms LHRH/2 h for 72 h), 2) be infused with LHRH (n = 11; 1.25 micrograms LHRH/h for 72 h), or 3) serve as controls (n = 16). Blood was collected at 20-min intervals for 8 h (0900 to 1700 h) from six heifers in each group on d 1, 2, 3 (during treatment), and on d 4 (during 8 h after terminating LHRH treatments). Heifers given LHRH had higher (P less than .01) LH concentrations than controls. Preovulatory-like LH surges occurred in three I, two P and no control heifers during treatment. Pulse frequencies of LH (no. LH pulses/8 h) were greater (P less than .001) for P heifers than for I and control heifers due to pulsatile LHRH treatment. Serum estradiol was higher (P less than .01) during treatment for LHRH-treated heifers than for controls. Serum follicle-stimulating hormone, cortisol, and progesterone were unchanged during treatment. High levels of cortisol on d 1 declined (P less than .001) to baseline by d 2. Characteristic progesterone rises or short luteal phases occurred within 10 d of treatment initiation in more (P less than .05) LHRH-treated heifers (I = 45%, P = 33%) than controls (6%), although days to first observed estrus and first ovulation were unaffected by treatments. Although both continuous and pulsatile administration of LHRH successfully induced LH and estradiol release as well as preovulatory-like LH surges in some heifers, earlier initiation of estrous cycles was not achieved. Estrous cycles appeared to be delayed by exposure to continuous LHRH infusions during the peripubertal period.     

Ovarian inhibition of pulsatile luteinizing hormone secretion in prepuberal holstein heifers

Fifteen prepuberal Holstein heifers were utilized to examine pulsatile luteinizing hormone (LH) secretion before and after ovariectomy. Heifers were ovariectornized at 3, 6 or 9 months of age (n=5/group) and scheduled for blood sampling at 1 week before, 1 week after and 4 weeks following ovariectomy. During each 8 hr sampling period (0600–1400 hr), blood samples (10 ml) were collected via indwelling jugular canulae at 10 min intervals. Prior to ovariectomy, mean plasma LH concentration and both number and amplitude of LH pulses per 8 hr sampling period were similar (P>.05) among age groups, and the absence of a pulsatile LH secretion profile was accompanied by a low mean LH concentration. Within 1 week after ovariectomy, both number of LH pulses and mean LH concentrations increased (P<.O1) in all age groups. Between 1 and 4 weeks after ovariectomy, both amplitude of LH pulses and mean LH concentrations increased (P<.O1) when the data from the three age groups were combined. We conclude that ovarian inhibition of pulsatile LH secretion is established by 3 months of age and is maintained through 9 months of age. In addition, the initial elevation mean plasma LH concentration is due to greater pulse frequency, while the subsequent rise in mean LH concentration reflects increased amplitude of LH pulses.     

Effects of body condition and estradiol on luteinizing hormone secretion in post-partum beef cows

An experiment was conducted to test the hypothesis that the effect of body fatness on LH pulsatility in post-partum cows is entirely independent of the negative feedback effects of ovarian steroids. Forty beef cows were fed in the last 100 d of gestation so that they achieved either a thin (mean score 1.97) or fat (mean score 2.79) body condition (0 to 5 scale) at calving and were fed after calving to maintain live weight and body condition. At 15 (sd 3.7) d post partum all cows were ovariectomised and half from each body condition score treatment group received a subcutaneous estradiol implant (+EST) while the remainder received no implant (−EST). At weeks 5 and 9 post-partum blood samples were collected via jugular catheter every 20 minutes for 10 hr on two consecutive d and on the third d cows were injected via the jugular vein with 2.5 μg GnRH. Blood samples were collected every 15 minutes for 1 hr before and 2 hr after GnRH injection. At 5 and 9 weeks the fatter cows had significantly higher mean LH concentrations, baseline LH concentrations, LH pulse amplitudes and pulse frequencies (P<0.01). Implantation with estradiol in both fat and thin cows reduced mean LH concentrations, baseline LH concentrations, LH pulse amplitudes and pulse frequencies (P<0.001). The lack of interaction between body condition and the presence or absence of estradiol implies that the effect of body condition on LH release is independent of ovarian steroid feedback mechanisms. Fat cows showed a greater release of LH in response to exogenous GnRH (P<0.01) than thin cows while implantation with estradiol in both fat and thin cows decreased (P<0.01) LH release. The pituitary responsiveness to GnRH with the −EST cows was greater at 9 compared to 5 weeks, but there was no difference with time in the +EST cows. However, there was no such interaction in endogenous LH pulse amplitude suggesting that in the absence of estradiol the magnitude of GnRH pulses declined with time post-partum.     

Induction of precocious puberty in heifers I: enhanced secretion of luteinizing hormone

In beef heifers weaned between 3 and 4 mo of age and fed a high-concentrate diet, approximately 50% reach puberty before 300 d of age (precocious puberty). The objectives of this experiment were 1) to determine whether precocious puberty could be induced experimentally by weaning heifers early and feeding a high-concentrate diet, and 2) to determine the dynamics of secretion of LH associated with precocious puberty. Crossbred Angus and Simmental heifer calves were weaned at 73 +/- 3 d of age and 115 +/- 3 kg of BW and fed a high-concentrate (60% corn; HI, n = 9) or control diet (30% corn; CONT, n = 9). Heifers were fed individually, and target BW gains were 1.50 and 0.75 kg/d for the HI and CONT treatments, respectively. Heifers were weighed every 2 wk. Blood samples were collected weekly and assayed for progesterone concentration to determine age at puberty. Serial blood samples were collected at 20-min intervals for 24 h at mean ages of 102, 130, 158, 172, 190, 203, 217, 231, and 259 d and assayed for LH concentration to evaluate the dynamics of secretion of LH. Heifers fed the HI diet exhibited greater BW gain (P < 0.01) than CONT heifers (1.27 +/- 0.05 vs. 0.85 +/- 0.05 kg/d, respectively). As a result, BW in the HI treatment was greater (P < 0.01) than in the CONT treatment by 188 d of age and remained different through the end of the experiment. Precocious puberty occurred in 8 of 9 heifers fed the HI diet and 0 of 9 heifers fed the CONT diet. Age at puberty was reduced in the HI (P < 0.01) compared with the CONT heifers (262 +/- 10 vs. 368 +/- 10 d of age, respectively). Body weight at puberty was also reduced in the HI (P < 0.05) compared with the CONT treatment (327 +/- 17 vs. 403 +/- 23 kg, respectively). Heifers attaining puberty during the experiment continued with subsequent luteal phases as evidenced by cyclic patterns of progesterone concentrations. Frequency of pulses of LH (pulses/24 h) increased with age (P < 0.01) for both treatments. Heifers in the HI treatment exhibited a greater number of pulses of LH (P < 0.01) than those in the CONT treatment by 190 d of age and in all subsequent collection periods (treatment x age, P < 0.05). Mean LH concentrations also increased with age (P < 0.01) for both treatments but did not differ between treatments. In conclusion, precocious puberty induced by early weaning and feeding of a high-concentrate diet is preceded by increasing frequency of pulses of LH.     

Seasonal differences in the parameters of luteinizing hormone release to exogenous gonadotropin releasing hormone in prepubertal Holstein heifers in Sapporo

Stress due to summer heat has adverse effects on reproduction in Holstein dairy cattle. Summer suppression of reproduction of Holsteins can pose an important economic problem, even in Hokkaido prefecture located in the northern region of Japan. Hokkaido is one of the most important dairy farming areas of Japan. This study is an attempt to clarify the seasonal differences in the parameters of luteinizing hormone (LH) response to exogenous gonadotropin releasing hormone (GnRH) in Sapporo, Hokkaido, Japan. A total of 12 prepubertal heifers received an injection with GnRH analogue intramuscularly in either May (n=4, May group), July (n=4, July group), or November (n=4, November group), and serial blood samples were collected to analyze the parameters of the LH response curve after GnRH injection. The parameters were as follows: the basal LH concentration, peak LH concentration, duration from the time of GnRH injection to the time of the peak LH concentration, and area under the LH response curve (AUC). There were no significant differences in the basal and peak LH concentrations or the AUC among the three groups. The July group reached the LH peak significantly (P<0.05) faster than the May group, but there was no significant difference with the November group. Therefore, the results of the present study do not demonstrate an effect of summer heat on the LH response to the exogenous GnRH in Holstein heifers.     

Increased luteinizing hormone secretion and ovarian function in heifers actively immunized against estrogen and progesterone

This study was designed to test the effects of active immunization against estrogen and progesterone on patterns of luteinizing hormone (LH) and follicle stimulating hormone (FSH) secretion, ovarian characteristics and growth rate of heifers. Heifers were randomly assigned to four treatments: 1) control injection (n = 10); 2) ovariectomy (n = 9); 3) immunization against estrogen (anti-E, n = 10); and 4) immunization against estrogen and progesterone (anti-E+P4, n = 10). Three booster immunizations were administered at 1, 1.5 and 6 mo after primary immunization. Progesterone antibody binding was 40% (34 fmol at 1:600 final dilution) in the anti-E+P4 heifers, and estradiol-17 beta binding was 35% (30 fmol) and 60% (52 fmol at 1:100 final dilution) in the anti-E+P4 and anti-E heifers, respectively, after the final immunization. Anti-E+P4 heifers had more pulses of LH and higher basal concentrations of LH than anti-E or control heifers (P less than .05). Concentrations of LH in anti-E+P4 heifers did not increase to concentrations found in ovariectomized heifers (P less than .05). Immunization against steroids did not alter the secretion of FSH. The number of large follicles (greater than 15 mm diameter) in anti-E+P4 and anti-E heifers was greater than in control heifers (P less than .05). Ovarian weight was increased in anti-E+P4 heifers (P less than .05). Average daily gain was not different among groups (P greater than .05). It was concluded that active immunization against estrogen and progesterone in heifers increased LH secretion and stimulated ovarian function.     

Effects of 17 beta-estradiol and diets varying in energy on secretion of luteinizing hormone in beef heifers

An experiment was conducted to test the hypothesis that 17 beta-estradiol (E2) would not suppress secretion of luteinizing hormone (LH) in heifers fed a diet limited in energy during the period before the onset of nutritionally induced anestrus. Sixteen of 20 heifers that had been exhibiting normal estrous cycles (20 mo of age, 409 +/- 6 kg body weight) were ovariectomized, and half of them were assigned at random to receive an E2 implant. The ovariectomized heifers were assigned at random to receive diets that contained low (L; 5.8 Mcal X animal-1 X d-1, n = 8) or high levels of energy (H; 20.0 Mcal X animal-1 X d-1, n = 8) for 100 d. The other four heifers remained intact and were fed the L-diet. The intact heifers were utilized to determine the status of reproductive function in animals fed the L-diet. Heifers lost body weight rapidly after initiation of feeding the L-diet. Heifers fed the L-diet then stabilized at a lighter weight until the latter part of the experiment. One of the four intact heifers fed the L-diet became anestrus near the end of the study. Mean concentrations of LH in blood serum increased linearly (P less than .05) in ovariectomized heifers fed the L- and H-diet. Mean concentration of LH in heifers fed the H-diet that were implanted with E2 was similar to ovariectomized heifers fed the H-diet that received no E2. Mean LH in serum of ovariectomized heifers implanted with E2 fed the L-diet was suppressed and remained low throughout the study. Frequency of pulses of LH in ovariectomized heifers fed the L-diet was less (P less than .01) than that in ovariectomized heifers fed the H-diet. Estradiol decreased the number of pulses of LH in heifers fed the L-diet. We conclude that dietary energy restriction in beef heifers has a direct action on the hypothalamo-pituitary axis to lower the number of pulses of LH in the absence of ovarian steroids. However, ovarian E2 appears to suppress further secretion of LH in heifers fed limited levels of dietary energy before the onset of nutritional anestrus occurs, therefore, our working hypothesis is rejected.     

Progesterone concentration,estradiol pretreatment,and dose of gonadotropin-releasing hormone affect gonadotropin-releasing hormone-mediated luteinizing hormone release in beef heifers

We examined whether progesterone (P₄)-induced suppression of LH release in cattle can be overcome by an increased dose of exogenous gonadotropin-releasing hormone (GnRH) or pretreatment with estradiol (E₂). In Experiment 1, postpubertal Angus-cross heifers (N = 32) had their 2 largest ovarian follicles ablated 5 d after ovulation. Concurrently, these heifers were all given a once-used, intravaginal P₄-releasing insert (CIDR), and they were randomly assigned to be given either prostaglandin F_2α (Low-P₄) or no treatment (High-P₄) at follicle ablation, and 12 h later. Six days after emergence of a new follicular wave, half of the heifers in each group (n = 8) were given either 100 or 200 μg of GnRH i.m. Plasma luteinizing hormone (LH) concentrations were higher in the Low- vs High-P₄ groups, and in heifers given 200 vs 100 μg of GnRH (mean ± SEM 15.4 ± 2.2 vs 9.1 ± 1.2, and 14.8 ± 2.1 vs 9.8 ± 1.4 ng/mL, respectively; P ≤ 0.01). Ovulation rate was higher (P = 0.002) in the Low-P₄ group (15/16) than in the High-P₄ group (6/16), but it was not affected by GnRH dose (P = 0.4). In Experiment 2, heifers (n = 22) were treated similarly, except that 5.5 d after wave emergence, half of the heifers in each group were further allocated to be given either 0.25 mg estradiol benzoate i.m. or no treatment, and 8 h later, all heifers were given 100 μg GnRH i.m. Both groups treated with E₂ (Low- and High-P₄) and the Low-P₄ group without E₂ had higher peak plasma LH concentrations compared to the group with high P₄ without E₂ (12.6 ± 1.8, 10.4 ± 1.8, 8.7 ± 1.3, and 3.9 ± 1.2 ng/mL, respectively; (P < 0.04)). However, E₂ pretreatment did not increase ovulation rates in response to GnRH (P = 0.6). In summary, the hypotheses that higher doses of GnRH will be more efficacious in inducing LH release and that exogenous E₂ will increase LH release following treatment with GnRH were supported, but neither significantly increased ovulation rate.     

Effect of age and body weight on plasma concentrations of luteinizing hormone in turkey hens before and after photostimulation

The independent effects of age and body weight (BW) on photostimulatory response in turkey breeder hens were studied by measuring changes in plasma luteinizing hormone (LH; ng/mL) before and 3 d after photostimulation. The study was conducted with hens from two BW groups at 24–25, 27–28, and 31–32 wk of age. There was approximately a 1-kg difference in BW between groups within an age. Six hens per BW and age group were cannulated (jugular vein) and serially sampled during each of two 6-hr periods. Samples were collected at 10-min intervals. The two sampling periods were the last 6 hr of the short-day photoperiod (SD) and the same period during the third long day after photostimulation (LD). The photostimulatory response (PR) or difference between the SD and LD baseline LH concentrations was greatest in the 24–25-wk-old hens. The PR was unaffected by hen BW at any age. The baseline LH concentration during the SD photoperiod declined as hens aged. After photostimulation, baseline LH and LH peak amplitude concentrations were higher in 24–25-wk-old hens compared with the older ages. The number of LH peaks increased after photostimulation, but there were no significant effects attributable to age or BW within an age. In conclusion, the PR was affected by hen age but not hen BW or BW within a particular age.     

Effects of steroid hormone treatment on mammary development in prepubertal heifers

The objective of this study was to determine the effects of steroid hormone implantation in heifer calves on the ability of mammary tissue to develop subsequently in organ culture. Twenty-four calves were paired by date of birth and assigned to groups (eight calves/group). At 4, 7, or 10 mo of age, calves were implanted subcutaneously (s.c.) with pellets containing cholesterol or cholesterol, 17β-estradiol, and progesterone for 9 or 18 d. The calves were euthanized and uteri and mammary glands were removed and weighed. Slices of mammary parenchymal tissue were incubated for 5 d at 37°C in a 50% O₂, 5% CO₂ humidified atmosphere in Waymouth’s 752/liter medium supplemented with insulin (5.0 μg/ml) or lactogenic hormone complex insulin (5.0 μg/ml), aldosterone (0.1 μg/ml), hydrocortisone (0.1 μg/ml), and prolactin (1.0 μg/ml) in the presence or absence of epidermal growth factor (EGF) (0.06 μg/ml) to promote lobulo-alveolar development. Tissue sections were stained and mounted on slides for morphologic and histologic analysis or prepared to evaluate expression of β-casein mRNA. There were no morphologic differences in slices from calf mammary tissues despite age, steroid hormone priming, or hormones used in tissue culture. The 4-mo-old calves required steroid priming followed by incubation of the tissue slices with the lactogenic complex with or without epidermal growth factor to induce cytological changes associated with lactogenesis but did not express β-casein mRNA. At 7 mo of age, steroid hormone priming was not necessary for induction of alveolar formation and secretion. Incubation of the tissue slices from 7-mo-old calves with the lactogenic complex was sufficient to induce alveolar formation and secretion. However, β-casein mRNA was not expressed. At 10 mo of age, exposure of tissue from calves to the lactogenic hormones caused histologic changes reminiscent of the ability to secrete milk regardless of hormone priming. However, estrogen and progesterone priming was necessary before incubation of the tissue slices with the lactogenic hormones to induce β-casein mRNA expression. When epidermal growth factor was added to the lactogenic hormone complex, β-casein mRNA expression decreased. These data support the concept that there is a sequential development of responsiveness of the mammary gland to various hormones. By 10 mo of age, prepubertal heifers reach a stage of maturity where steroid hormone priming followed by incubation of tissue slices with the lactogenic hormones is sufficient to induce both structural and functional differentiation.     

Release of luteinizing hormone after administration of naloxone in pre- and peripuberal heifers.

This study was conducted to investigate regulation of LH release by opioid peptides during puberal development in beef heifers. Fourteen heifers were randomly assigned to receive naloxone (opioid antagonist) i.v. at dosages of either 1 mg.kg BW-1.wk-1 (Dose 1) or .25 mg.kg BW-1.wk-1 (Dose 2) for 13 wk or until puberty. Blood was sampled (one sample every 15 min) 6 h before (prenaloxone) and 2 h after naloxone administration. Two hours after naloxone administration, GnRH (10 ng/kg BW) was administered and blood was sampled for 1 h. Nine heifers attained puberty during the study. There were no differences between naloxone dosage groups for any measured variables. Therefore, heifers were grouped dependent on the attainment of puberty. Prenaloxone concentrations of serum LH and LH pulse frequency were normal for prepuberal heifers. Serum LH concentrations increased within 30 min after naloxone 135 of 139 times it was administered (P less than .05). Serum LH concentrations during the hour after naloxone were higher (P less than .05) than those during the hour before naloxone in both puberal and nonpuberal heifers. In puberal heifers, serum LH pulse height during the hour after naloxone was greater (P less than .02) at 5 wk before puberty and lower (P less than .02) the week before puberty than at other times during the trial. There was no effect of week on serum LH pulse height after naloxone in heifers that failed to attain puberty during the study. Response of LH to GnRH was similar between groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Associated luteinizing hormone-releasing hormone and luteinizing hormone secretion in ovariectomized gilts.

The secretion of luteinizing hormone-releasing hormone (LHRH) and its temporal association with pulses of luteinizing hormone (LH) was examined in ovariectomized prepuberal gilts. Push-pull cannulae (PPC) were implanted within the anterior pituitary gland and LHRH was quantified from 10 min (200 microliters) perfusate samples. Serum LH concentrations were determined from jugular vein blood obtained at the midpoint of perfusate collection. Initial studies without collection of blood samples, indicated that LHRH secretion in the ovariectomized gilt was pulsatile with pulses comprised of one to three samples. However, most pulses were probably of rapid onset and short duration, since they comprised only one sample. Greater LHRH pulse amplitudes were associated with PPC locations within medial regions of the anterior pituitary close to the median eminence. In studies which involved blood collection, LH secretion was not affected by push-pull perfusion of the anterior pituitary gland in most gilts, however, adaptation of pigs to the sampling procedures was essential for prolonged sampling. There was a close temporal relationship between perfusate LHRH pulses and serum LH pulses with LHRH pulses occurring coincident or one sample preceding serum LH pulses. There were occasional LHRH pulses without LH pulses and LH pulses without detectable LHRH pulses. These results provide direct evidence that pulsatile LHRH secretion is associated with pulsatile LH secretion in ovariectomized gilts. In addition, PPC perfusion of the anterior pituitary is a viable procedure for assessing hypothalamic hypophyseal neurohormone relationships.     

Influence of insulin and energy intake on ovulation rate, luteinizing hormone and progesterone in beef heifers

Ovarian and gonadotropin responses to insulin and energy restriction were investigated in a 2 X 2 factorial experiment using 2-yr-old Brangus heifers. Thirty heifers were paired by weight and body condition, then assigned to treatment groups receiving 75 (LE) or 180% (HE) of NRC recommendations for dietary energy for maintenance. Diets were adjusted weekly to maintain daily .25 to .5 kg weight loss or 0 to .25 kg weight gain, respectively. On d 10 of the first estrous cycle subsequent to the initial 45 d of feeding, heifers within each dietary group were allocated to receive twice daily infusions of either 40 U insulin (I) or saline (C). Infusions began at 5 and 10 h postprandial and were given in six boluses, 20 min apart. Infusions continued daily until d 20 or estrus, whichever occurred first. On d 11, blood samples were collected at 15-min intervals for 12 h to determine luteinizing hormone (LH) and insulin concentrations. On d 16 to 20, twice daily im injections of 1 mg follicle stimulating hormone (FSH) were administered. Heifers were ovariectomized on d 11 after estrus. Number of corpora lutea (CL) in LE-I heifers was greater (P less than .05) in LE-C, HE-C or HE-I. Total CL weight (g) per heifer was greater (P less than .05) in HE-C and LE-I heifers than in LE-C. Individual CL wt was heavier in HE than in LE heifers (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of castration on plasma luteinizing hormone concentrations in prepubertal boars

Mean concentrations and the occurrence of pulsatile release of luteinizing hormone (LH) were determined in 14-wk-old crossbred boars (50.5 +/- 1.5 kg) after bilateral or unilateral castration at 10 wk of age. Blood was collected at 10-min intervals for 5 h. Then gonadotropin releasing hormone (GnRH; 40 micrograms) was given and sampling was continued at 5-min intervals for 1 h. Compared with intact boars, bilateral castration increased (P less than .001) mean LH (982 +/- 56 vs 389 +/- 56 pg/ml), pulsatile releases of LH (7.0 +/- .6 vs 2.0 +/- .6 pulses/5 h) and LH pulse amplitude (617 +/- 29 vs 360 +/- 58 pg/ml). Unilaterally castrated boars did not differ from intact boars in any of the above measures of LH secretion. Testis weight increased more between 10 and 14 wk of age in the unilateral castrates than in the intact boars (432 +/- 42 vs 245 +/- 34%; P less than .05). Thus, compensatory hypertrophy occurred within 4 wk of castration. Plasma testosterone was lower for bilateral castrates than for intact animals (.1 +/- .8 vs 3.6 +/- .9 ng/ml; P less than .05) while unilateral castrates (3.8 +/- 1.0 ng/ml) and intact boars did not differ. Plasma estradiol concentrations in bilateral and unilateral castrates were not different from levels found in intact boars (1.8 +/- 1.8, 8.8 +/- 2.1 and 6.0 +/- 1.8 pg/ml, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of nutrition on serum concentrations of progesterone, luteinizing hormone and estrous behavior in dairy heifers

Thirty postpubertal Holstein heifers were used to investigate the influence of dietary intake on 1) serum concentrations of progesterone (P4) and luteinizing hormone (LH) during diestrus and 2) estrous behavior. Heifers were randomly allotted to receive either 80, 100 or 120% of the National Research Council (NRC) requirements for energy, protein and dry matter intake for 139 d. Heifers were fed their respective diets in groups in outdoor lots for 114 d at which time individual feeding of diets was initiated in a stanchion barn. Estrus was synchronized with two injections of prostaglandin (PG) so that on d 138 of dietary treatment the heifers were in diestrus (means = d 8 of estrous cycle). Also, on d 138, blood samples were collected via jugular cannula at 20-min intervals for 12 h beginning at 0630. After collection of the 37th blood sample, all heifers were treated with PG and additional blood samples were collected at 1-h intervals for 24 h. Immediately thereafter, all heifers were moved to a common outdoor lot for 72 h of continuous observation for estrous behavior. Weight gains during the experimental period differed among groups and were 46 +/- 5, 83 +/- 5 and 113 +/- 4 kg for the heifers fed 80, 100 and 120% of the NRC requirements, respectively. Mean serum concentrations of P4 for the 80% NRC group (4.6 +/- .6 ng/ml) tended to be lower than both the 100% (5.9 +/- .6 ng/ml) and 120% (6.0 +/- .5 ng/ml) NRC groups, respectively. Mean serum concentrations of LH, number of LH secretory peaks/12 h and LH peak amplitude were similar among all NRC groups. Rate of decline in serum P4 after PG was also similar among all NRC groups.(ABSTRACT TRUNCATED AT 250 WORDS)