Ovine luteinizing hormone: isoforms in the pituitary during the follicular and luteal phases of the estrous cycle and during anestrus.

Pituitaries were collected from late follicular phase (n = 5), mid-luteal phase (n = 5), and anestrous ewes (n = 4) to assess changes in intrapituitary LH heterogeneity at selected reproductive states. After homogenization, an aliquot of each pituitary extract was desalted by flow dialysis against water and chromtofocused on a pH 10.5 to 4.0 gradient. Concentrations of LH in pituitary extracts and chromatofocusing fractions were determined by RIA. The LH in pituitary extracts resolved into 13 isoforms during chromatofocusing, which were coded with letters beginning with the most basic isoform. Follicular and mid-luteal phase ewes exhibited similar distributions of intrapituitary LH among its isoforms. Relative to follicular and luteal phase ewes, anestrous ewes had lower percentages of isoforms D and E as well as higher percentages of isoforms G, H, J and K. Isoform F, the predominant molecular form of LH, constituted a similar percentage in all treatment groups (P > .05). Thus, the distribution of intrapituitary LH among its isoforms did not change significantly between the mid-luteal and follicular phases of the estrous cycle, but higher percentages of the weakly basic and acidic forms of LH were present during anestrus. These observations suggest that intrapituitary LH heterogeneity changes minimally throughout the estrous cycle of ewes during the breeding season.     

Ultrasonographic appearance of the ovaries of dogs during the follicular and luteal phases of the estrous cycle.

Ultrasonography of the ovaries of 10 bitches was performed daily, using a 7.5-Mhz transducer with a built-in stand-off pad, from the onset of proestrus until the onset of metestrus. Ovarian size, shape, location, echogenicity, follicular development, and apparent ovulation were monitored. Blood samples were collected twice daily for luteinizing hormone determination and daily for progesterone determination. Vaginal smears were made daily for cytologic evaluation. Ultrasonograms were evaluated independent of hormonal and cytologic data, and the day of ovulation was noted. Initially, the ovaries were uniform and had an echogenicity that was equal to or slightly greater than that of the renal cortex. Follicles appeared as focal hypoechoic to anechoic rounded structures. Ovaries were easier to identify as follicular development progressed. Ovarian size increased with time. Apparent ovulation was characterized by a decrease in number of follicles seen from 1 day to the next, but 1 or more follicles remained in at least 1 ovary of 7 of 10 bitches. The ovaries had an oval shape that became rounded after ovulation. At some time after ovulation, all bitches had cystic (anechoic) structures indistinguishable from follicles. These structures increased in echogenicity and decreased in size with time and may have been follicles that did not ovulate, corpora hemorrhagica, fluid-filled corpora lutea, or cystic luteinized follicles. Time of ovulation determined by ultrasonography paralleled that predicted on the basis of hormonal data in 9 of 10 bitches and with cytologic findings in all bitches.     

Plasma hormone concentrations after administration of dexamethasone during the middle of the luteal phase in cows

The effect of glucocorticoids on gonadal steroid and gonadotropic hormone concentrations and subsequent follicular activity in cows undergoing normal estrous cycles was evaluated by administration of dexamethasone (DXM) during the middle of the luteal phase. Seven cows were given physiologic saline solution twice daily from day 13 to day 17 of the estrous cycle (control experiment). During the next estrous cycle, cows were administered DXM (2 mg, IM) twice daily on days 13 through 17. Plasma specimens obtained twice daily throughout the control and DXM-treatment cycles were assayed for progesterone and estradiol concentrations. The appearance of estrus after DXM treatment was delayed until days 23 to 25 in 3 cows and was not seen by day 35 in the other 4 cows, compared with mean (+/- SD) cycle length of 22.4 +/- 3.2 in cows during the control experiment. Progesterone concentration remained significantly (P less than 0.01) high on days 19 to 23, whereas estradiol values failed to increase (P less than 0.05) on days 19 and 20 after treatment with DXM. Blood samples were obtained at 15-minute intervals for 12 hours to compare (by analysis of covariance) the effect of DXM treatment on plasma hormone concentrations on day 15 of each cycle with those of day 10. Compared with values during the control experiment, a significant (P less than 0.05) decrease was observed in the size of the pulses of luteinizing hormone (LH) and estradiol, although the number of pulses of each hormone per 12 hours was not affected when cows were given DXM. Baseline concentrations of LH and estradiol were not altered by type of treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

Follicle stimulating hormone pattern and luteal function in ewes receiving bovine follicular fluid during three stages of the estrous cycle

The objectives of this study were to determine 1) the ability of charcoal-extracted bovine follicular fluid (bFF) to suppress endogenous follicle stimulating hormone (FSH) at various stages of the estrous cycle and 2) the effects of suppression of FSH on luteal function and lengths of the current and subsequent estrous cycles. Twenty-six mature ewes were assigned randomly to receive 5 ml of either bFF or saline, subcutaneously, at 8-h intervals on d 1 through 5 (bFF n = 6; saline n = 3), d 6 through 10 (bFF n = 6; saline n = 3) or d 11 through 15 (bFF n = 6; saline n = 2) of the estrous cycle (d 0 = estrus). Blood was collected daily beginning at estrus and continued until the third estrus (two estrous cycles) or 40 d; more frequent samples were collected 2 h prior to initiation of treatment (0600), hourly for the first 8 h of treatment, then every 4 h until 0800 on the first day after treatment, and finally at 1600 and 2400 on that day. Plasma concentrations of FSH were lower (P less than .001) in bFF-treated than in saline-treated ewes. Treatment with bFF reduced (P less than .05) plasma concentrations of progesterone during the current but not during the subsequent estrous cycle. Treatment with bFF did not affect plasma concentrations of estradiol-17 beta. Administration of bFF on d 11 through 15 of the estrous cycle lengthened the interval from the decline in progesterone to estrus and the inter-estrous interval by approximately 3 and 4 d, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cortisol and luteinizing hormone after adrenocorticotropic hormone administration to postpartum beef cows

Cortisol and luteinizing hormone (LH) were measured in serum after the administration of adrenocorticotropic hormone (ACTH) to suckled (S) and nonsuckled (NS) beef cows. Blood was sampled on 2 consecutive days every 2 weeks for four bleeding periods starting 14 days after calving. Cows were injected with 200 IU ACTH or saline in a 2-day switchback design. Serum was collected before ACTH or saline injection and at 30-min intervals thereafter for 8 hours. Average cortisol concentrations in serum were similar in S and NS cows (6.4 +/- .6 and 6.1 +/- .8 ng/ml, respectively) after saline. Average cortisol concentrations in serum collected during an 8-hr period after ACTH on days 14, 28, 42 and 56 postpartum were 24.7 +/- 2.4, 31.8 +/- 3.5, 36.4 +/- 4.2 and 40.7 +/- .5 ng/ml, respectively, for S cows, and 31.1 +/- 2.9, 44.7 +/- 5.2, 45.0 +/- 5.7 and 46.0 +/- 5.4 ng/ml, respectively, for NS cows. Cortisol response to ACTH, measured as area under the response curve, was greater (P less than .05) in NS than in S cows. Amount of cortisol released by 200 IU ACTH was maximal by days 28 to 29 postpartum in NS cows, but the response increased gradually between days 14 to 15 and days 56 to 57 in S cows. overall, LH in serum averaged .55 +/- .08 ng/ml for S cows and .92 +/- .06 ng/ml for NS cows after saline, and .49 +/- .07 ng/ml for S cows and .94 +/- .06 ng/ml for NS cows after ACth. Although mean and peak serum LH concentrations did not differ between cows given ACTH and those given saline, the number of LH peaks and the number of cows having LH after saline. Mean serum LH concentrations were lower (P less than. 05) in S than in NS cows at 28 days postpartum. The number of LH peaks was lower (P less than .05) and the magnitude of the largest LH peak tended to be lower (P less than .06) in S cows at all sampling periods.     

Effect of lipoproteins and luteinizing hormone on progesterone production by large and small luteal cells throughout the porcine estrous cycle

Large and small cells were isolated from porcine corpora lutea (CL) on d 10, 15 or 18 of the estrous cycle. They were incubated 13 to 16 h in cholesterol- and serum-free media and then supplied with 0, 10, 50 or 100 micrograms of either porcine low density lipoprotein (LDL) or porcine high density lipoprotein (HDL). Each dose was supplemented with 0, 10, 50 or 100 ng of porcine LH. Media progesterone (P4) content was assessed immediately before and 2 and 24 h after addition of lipoproteins and LH. Production of P4 by large cells always exceeded that of small cells. Day 10 large cells were stimulated by LDL, unaffected by LH, and either inhibited or unaffected by HDL. No treatment affected d 10 small cells. Day 15 large cells and small cells were stimulated by both lipoproteins (LDL greater than HDL). The large cells were stimulated to a small extent by LH at 2 h (P less than .05). Large cells could be isolated from only two of five preparations of d 18 CL. Day 18 small cells produced a small quantity of P4 in a response which qualitatively resembled that of d 15 small cells. Cell type, cycle stage and lipoprotein (particularly LDL) were the major effectors of P4 production. The minimal response to LH supports the theory of autonomy of the porcine CL with respect to P4 production. Days 10 and 15 bracket the period of commitment to luteolysis, and the nature of P4 production by each cell type changed over that period.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effect of active immunization against inhibin on gonadotropin secretions and follicular dynamics during the estrous cycle in cows

The hypothesis of the present study is that active immunization of cows against inhibin would neutralize endogenous inhibin, increase circulating levels of follicle stimulating hormone, and subsequently affect follicular dynamics and the ovulation rate during the estrous cycle. Thirteen cows were immunized against inhibin alpha-subunit and, 6 cows were immunized with a placebo. Both groups were given 4 booster immunizations 7, 14, 21, and 34 weeks after the primary injection. Ovaries were examined daily after the 2nd, 3rd, and 4th booster immunizations by transrectal ultrasonography for 25 days. After the 4th booster immunization, blood samples were collected daily for one complete estrous cycle to measure FSH and LH. The results showed that the immunized cows generated antibodies against inhibin, and that they had higher FSH levels compared with the controls. The number of follicular waves during the estrous cycle was higher in the immunized cows (3 or 4 waves) than in the controls (2 or 3 waves). Moreover, the immunized cows had a greater number of follicles during the estrous cycle compared with the control cows. The maximum number of follicles was 14.8 +/- 1.7 vs 5.4 +/- 0.2 in inhibin-immunized and control cows, respectively, during the first follicular wave and 13.9 +/- 1.9 vs 5.6 +/- 0.7, respectively, during the ovulatory wave. Multiple ovulations were increased in the immunized cows. However, the ovulation rate varied greatly in the immunized animals. In conclusion, immunization against inhibin increased FSH secretions during the estrous cycle in the cows. Moreover, the immunized cows had a greater number of follicular waves during the estrous cycle and a greater number of follicles, and this could be used as a potential source of oocytes for use in IVF/embryo transfer programs.     

Effects of different five-day progesterone-based synchronization protocols on the estrous response and follicular/luteal dynamics in dairy cows

This study compared the responses shown by lactating dairy cows to four different P4-based protocols for AI at estrus. Cows with no estrous signs 96 h after progesterone intravaginal device (PRID) removal were subjected to fixed-time AI (FTAI), and their data were also included in the study. In Experiment I, follicular/luteal and endometrial dynamics were assessed every 12 h from the beginning of treatment until AI. The estrous response was examined in Experiment II, and fertility was assessed in both experiments. The protocols consisted of a PRID fitted for five days, along with the administration of different combinations of gonadotropin releasing hormone (GnRH), equine chorionic gonadotropin and a single or double dose (24 h apart) of prostaglandin F_2α. In Experiment I (40 cows), animals receiving GnRH at the start of treatment showed a significantly higher ovulation rate during the PRID insertion period while estrus was delayed. In Experiment II (351 cows), according to the odds ratios, cows showing luteal activity at the time of treatment were less likely to show estrus than cows with no signs of luteal activity. Treatment affected the estrous response and the interval from PRID removal to estrus but did not affect conception rates 28–34 days post AI. Primiparous cows displayed a better estrous response than multiparous cows. Our findings reveal acceptable results of 5-day P4-based protocols for AI at estrus in high-producing dairy cows. Time from treatment to estrus emerged as a good guide for FTAI after a 5-day P4-based synchronization protocol.     

Influence of exogenous adrenocorticotropic hormone on estrous behavior in cattle.

A two-trial experiment was conducted to determine the influence of ACTH on estrous behavior in cattle. In Trial 1, Holstein heifers (n = 20) received an injection of prostaglandin F2 alpha (PGF) during a synchronized diestrus and 30 h later were allotted randomly to receive (i.m.) either 1) 4 mL of gelatin (Veh) or 2) 320 units of ACTH in 4 mL of gelatin (ACTH). Eleven days after the PGF injection, all heifers were again injected with PGF, and they received either Veh or ACTH to complete a cross-over design. Treatment with ACTH decreased (P less than .05) the duration of estrus (12.0 +/- 1.9 vs 18.0 +/- 1.6 h for Veh) and increased (P less than .001) the interval to estrus after PGF injection (62.9 +/- 2.6 vs 43.7 +/- 2.2 h for Veh). Peak serum concentrations of progesterone (P4) and cortisol (C) were elevated (P less than .001) after ACTH compared with Veh. In Trial 2, ovariectomized Holstein cows (n = 12) were injected (i.m.) with .5 mg of estradiol benzoate (EB) and, 10 h later, were allotted randomly to receive (i.m.) either 1) 4 mL of gelatin (Veh) or 2) 320 units of ACTH in 4 mL of gelatin (ACTH). Seven days after the initial EB injection, all cows were again injected with .5 mg of EB and, 10 h later, received either Veh or ACTH to complete a cross-over design. Treatment with ACTH decreased (P less than .01) the proportion of cows in estrus (2/12 vs 11/12 for Veh) and increased (P less than .01) peak serum concentrations of P4 and C.(ABSTRACT TRUNCATED AT 250 WORDS)     

Anovulation and plasma hormone concentrations after administration of dexamethasone during the middle of the luteal phase in sows undergoing estrous cycles

The effect of glucocorticoids on early follicular growth in sows undergoing normal estrous cycles was evaluated by administration of dexamethasone during the middle of the luteal phase. Plasma specimens were obtained for measurement of luteinizing hormone (LH), follicle-stimulating hormone (FSH), progesterone, and estradiol-17 beta concentrations. Fifteen sows were used. Control sows (n = 5) were given physiologic saline solution twice daily from day 9 to day 14 of the estrous cycle. Sows of the second group (n = 5) were given dexamethasone (30 micrograms/kg of body weight, IM) similarly, and those of the third group (n = 5) were given dexamethasone plus gonadotropin-releasing hormone (GnRH; 50 micrograms at 6-hour intervals, IV). Plasma specimens, obtained twice daily from day 8 through day 26, indicated that progesterone production and luteal regression were not inhibited by any of the 3 treatment regimens. Although preovulatory plasma estradiol concentration increased in control sows, such was not observed in the sows treated with dexamethasone or dexamethasone plus GnRH (P less than 0.01). Ovulation, with formation of corpora lutea, occurred in gilts given saline solution. Dexamethasone administration resulted in persistence of 19 to 41 follicles/ovary (2 to 4 mm in diameter), and dexamethasone-plus-GnRH treatment resulted in 6 to 18 follicles/ovary (5 to 6 mm in diameter). Plasma was obtained at 15-minute intervals for 12 hours to compare the effect of treatment on hormone concentrations on day 12 of the estrous cycle with the values on day 8.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in the peripheral concentrations of inhibin, follicle-stimulating hormone, luteinizing hormone, progesterone and estradiol-17beta during turnover of cystic follicles in dairy cows with spontaneous follicular cysts

Several studies have clarified that the follicular cysts degenerate and are replaced by newly growing follicles that develop into new follicular cysts without ovulation, i.e., turnover of ovarian follicular cysts in cows. However, the relativity of endocrinological changes, including the inhibin profile during turnover of spontaneous follicular cysts in dairy cows, is still unclear. In the present study, the relationship between turnover of follicular cysts and changes in the peripheral blood concentrations of progesterone (P), estradiol-17beta (E(2)), luteinizing hormone (LH), follicle stimulating hormone (FSH) and inhibin were examined in lactating dairy cows. Five cows diagnosed with follicular cysts (follicles of more than 25 mm in diameter in the absence of a corpus luteum) were investigated. Their ovarian dynamics were monitored using ultrasonography, and blood samples were collected at 2- or 3- day intervals throughout the experiment. The day when a follicle fated to become a follicular cyst reached more than 8 mm in diameter was defined as the start of a cystic follicular wave. Four of the 5 cows exhibited a similar patterns of cystic follicular changes and hormone profiles. The data from the 4 cows was used for analysis of the relationships between turnover of cystic follicles and the hormone profiles. Two or three new cystic follicular waves occurred in each cow during the experimental period. The mean diameter of the cystic follicles was more than 25 mm 13 to 15 days after the start of the cystic follicular wave, and it began to decrease 1 to 6 days before the start of the subsequent cystic follicular wave. The levels of E(2) and inhibin tended to decrease for 7 to 9 days before the start of a new cystic follicular wave and to increase concomitantly with new follicular cyst growth. The levels of FSH rose for 1 to 3 days before the start of a new cystic follicular wave. The present study clarified the relationship between FSH and inhibin during turnover of spontaneous follicular cysts in dairy cows and found that it was very similar to previous results for cows. The present results suggest that an increase in FSH secretion following a reduction in inhibin secretion triggers turnover of cystic follicles in cows with spontaneous follicular cysts.     

Steroid hormone and luteinizing hormone concentrations in the anestrous sow.

This investigation characterized serum concentrations of luteinizing hormone (LH), estradiol-17 beta (E2), progesterone (P4) and cortisol (C) in anestrous sows. Twenty-two sows that had not returned to estrus within 45 days after weaning (anestrous sows), and ten sows that had returned to estrus within seven days following weaning (cyclic sows) were nonsurgically fitted with indwelling jugular vein cannulae. Blood samples were collected at 6 h intervals for seven days and at 15 min intervals for 8 h on the fifth day after cannulation. Serum LH concentrations were determined in all samples, while C, E2 and P4 levels were quantitated in serum collected at 6 h intervals. Serum P4 concentrations in anestrous sows were consistently less than 0.5 ng/mL, and E2 levels ranged from 10 to 19 pg/mL. Concentrations of LH remained less than 1.0 ng/mL in anestrous sows, whereas a preovulatory LH surge was observed in five of ten cyclic sows. There was a circadian rhythm in mean C levels with C peaks occurring at 0600 or 2400 h and nadir levels observed at 1200 and 1800 h. Few differences in C levels were detected between anestrous and cyclic sows. It was evident that anestrous sows did not exhibit cyclic or predictable variations in steroid hormone concentrations. Unfortunately, the results of this study failed to elucidate the endocrine pathogenesis of the anestrous sow.     

Plasma cortisol and progesterone responses to low doses of adrenocorticotropic hormone in ovariectmized lactating cows

The objective of this study was to describe the responses of the plasma progesterone and cortisol concentrations in ovariectomized lactating cows to low doses of adrenocorticotropic hormone (ACTH). The estrous cycles in 3 lactating cows were synchronized, and the cows were ovariectomized in the luteal phase. ACTH challenge tests were conducted at doses of 3, 6, 12 and 25 IU. Blood samples were collected at 30 min intervals, and the plasma progesterone and cortisol concentrations were analyzed by EIA. A concomitant rise in plasma progesterone and plasma cortisol was observed in cows treated with 12 IU or higher doses of ACTH. Significant increments in the plasma cortisol concentrations were observed at all doses of ACTH. The means (+/- SE) of the peak plasma progesterone concentrations after the 3, 6, 12 and 25 IU ACTH challenge tests were 0.6 +/- 0.1, 1.3 +/- 0.4, 1.5 +/- 0.3 and 2.4 +/- 0.3 ng/ml, respectively. The means of the peak plasma cortisol concentrations in the 3 cows after the ACTH challenge were 14.0 +/- 1.5, 17.0 +/- 2.5, 23.3 +/- 3.0, and 33.3 +/- 7.0 ng/ml, respectively. The effects of the doses, time after treatment, and their interaction on the plasma progesterone concentrations after the ACTH challenge were significant (P<0.01). Likewise, the effects of the doses, time after treatment, and their interaction on the plasma cortisol concentrations after the ACTH challenge were significant (P<0.01). The mean AUC values for the plasma progesterone and cortisol concentrations after the ACTH treatments were also significantly affected by the dose of ACTH (P<0.01 and P<0.05, respectively). A significantly positive correlation was obtained between the peak plasma progesterone and cortisol concentrations after different doses of ACTH (r=0.7, P<0.05). The results suggest that lactating dairy cows are capable of secreting a significant amount of adrenal progesterone, reaching up to the minimal concentration necessary to cause suppression of estrus in response to 12 IU ACTH (P<0.01). The concomitant plasma cortisol concentration was 23.3 ng/ml.