Pituitary receptors for GnRH and estradiol, and pituitary content of gonadotropins in beef cows. II. Changes during the postpartum period

Pregnant beef heifers (n = 24) were assigned randomly to four groups and slaughtered at day 1, 15, 30 or 45 postpartum. The day prior to slaughter blood samples were taken from each cow every 15 min for 8 hr. The anterior pituitary gland, preoptic area (POA) and medial basal hypothalamus (HYP) were collected from each cow. Contents of gonadotropin-releasing hormone (GnRH) in extracts of POA and HYP, and luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in extracts of anterior pituitary were quantified by radioimmunoassay. In the anterior pituitary gland, membrane receptors for GnRH were quantified by a standard curve technique and cytosolic receptors for estradiol were quantified by saturation analysis. Concentrations of LH, FSH and prolactin in serum were quantified by radioimmunoassay. Only one cow of eight had a pulse of LH during the 8 hr bleeding period on day 1 postpartum. This increased to 8 pulses in 6 cows on day 30 postpartum. Contents of GnRH in POA (15.0 +/- 3.2 ng) and HYP (14.0 +/- 2.0 ng) did not change significantly during the postpartum period. Pituitary content of LH was low following parturition (.2 +/- .1 mg/pituitary) and increased significantly through day 30 postpartum (1.2 +/- .1 mg/pituitary). Pituitary content of FSH did not change over the postpartum period. Receptors for both GnRH (.9 +/- .2 pmoles/pituitary) and estradiol (5.0 +/- .9/moles/pituitary) were elevated on day 15 postpartum, possibly increasing the sensitivity of the anterior pituitary gland to these hormones and leading to an increased rate of synthesis of LH that restored pituitary content to normal by day 30 postpartum.     

Pituitary concentrations of gonadotropins and receptors for GnRH in suckled beef cows at various intervals after calving

Mature beef cows were slaughtered at 5 (n = 6), 10 (n = 6), 20 (n = 6) or 30 (n = 5) d after calving to identify endocrine events that may affect the duration of postpartum anestrus. Additional cows (n = 6) were slaughtered 12 to 14 d after their first postpartum estrus (luteal phase cows). Anterior pituitary concentrations of luteinizing hormone (LH) were low at d 5 (383 +/- 69 micrograms/g), averaged 445 +/- 103 and 682 +/- 207 micrograms/g at d 10 and 20, respectively, and were elevated (P less than .05) by d 30 (1,097 +/- 174 micrograms) to a concentration similar to luteal phase cows (1,208 +/- 148 micrograms/g). Concentrations of follicle-stimulating hormone (FSH) averaged 12.4 +/- 1.1, 9.6 +/- 2, 8.6 +/- 1.8 and 7.4 +/- 3.3 mg/g at d 5, 10, 20 and 30, respectively. Affinity (1.6 +/- .2 X 10(9) M-1) of anterior pituitary receptors for the GnRH (gonadotropin-releasing hormone) analog (DAla6; des-Gly10, [D-Ala6]-LH-RH ethylamide) and weights (2.1 +/- .1 g) of the anterior pituitaries did not differ among groups (P greater than .05). Number of receptors for GnRH averaged 37 +/- 7, 39 +/- 9, 25 +/- 5 and 23 +/- 5 X 10(-14) M/mg protein at d 5, 10, 20 and 30, respectively. Anterior pituitaries from luteal phase cows contained 22 +/- 2 X 10(-14) M/mg protein of receptors for GnRH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of estradiol,progesterone, and nutrition on concentrations of gonadotropins and GnRH receptors,and abundance of mRNA for GnRH receptors and gonadotropin subunits in pituitary glands of beef cows

Nutritionally induced anovulatory cows (n = 28) were used to determine the effect of steroids on regulation of synthesis and secretion of gonadotropins. Anovulatory cows were ovariectomized and received intravaginal inserts containing estradiol (E2), progesterone (P4), E2 and P4 (E2P4), or a sham intravaginal insert (C) for 7 d. Concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were quantified in serum and E2 and P4 were quantified in plasma. Cows were exsanguinated within 1 to 2 h after removal of intravaginal inserts and pituitary glands were collected and stored at -80 degrees C until messenger ribonucleic acid (mRNA) for gonadotropin-releasing hormone receptor (GnRH-R) and gonadotropin subunits, pituitary content of GnRH-R, and LH and FSH were quantified. Pituitary glands from five proestrous cows were harvested to compare gonadotropin characteristics between ovariectomized, anovulatory cows and intact cows. Plasma concentrations of E2 were greater (P < 0.05) in E2-treated cows than in sham-treated cows. Concentrations of P4 were greater (P < 0.05) in cows treated with P4 than in sham-treated cows. Mean serum concentrations of LH and FSH were not significantly influenced by steroid treatments. However, frequency of LH pulses of ovariectomized, nutritionally induced anovulatory cows was increased (P < 0.05) by treatment with E2 and amplitude of LH pulses was greater (P < 0.05) in cows treated with E2 or P4 than in cows treated with E2P4 or sham-treated. Quantity of mRNA for LHbeta in the pituitary gland was greater when cows were treated with P4. Concentrations of LH in the pituitary gland were not affected by steroid treatments; however, pituitary concentrations of FSH were less (P < 0.1) in E2 cows than in sham-treated cows. The number of GnRH-R was increased (P < 0.05) in cows treated with E2, but P4 treatment did not influence the number of GnRH-R. Abundance of mRNA for GnRH-R, common alpha-subunit, and FSHbeta were not affected by treatments. Pituitary concentrations of LH were greater (P < 0.05) and concentrations of FSH were less (P < 0.05) in proestrous cows than in ovariectomized, anovulatory cows treated with or without steroids. Abundance of mRNA for GnRH-R, common alpha-subunit, LHbeta and FSHbeta were similar for proestrous and anovulatory cows. We conclude that treatment of nutritionally induced anovulatory cows with progesterone and estradiol may cause pulsatile secretion of LH.     

Changes in hormonal profiles during the estrous cycle in old lactating beef cows

Patterns of concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), progesterone (P4) and estradiol-17 beta (E2) during an estrous cycle were compared between 15 lactating beef cows 5 to 7 years of age (young) and 15 cows greater than or equal to 12 years of age (old). Length of estrous cycle did not differ between young and old cows (P = .06). No differences due to age were found for LH. Patterns of concentrations of P4 during the first 15 days of the cycle, of FSH during days 6 through 12 and of E2 during the follicular phase differed with age (P less than .05). An earlier (P less than .025) midcycle elevation of FSH was associated with an earlier rise and greater concentration of E2 (P less than .05) during the follicular phase in old than in young cows. Differences in FSH and P4, although subtle, were consistent with an earlier or more advanced follicular development in old cows, leading to greater secretion of E2 from the preovulatory follicle.     

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

Adenohypophyseal concentrations of LHRH receptors, pituitary content of LH and FSH, and plasma concentrations of LH were determined in thirty Hereford, Angus or Hereford-Angus heifers that were randomly assigned by breed and weight to five periods including day 3 of the estrous cycle (CY), pregnant day 120 (P120), 200 (P200), 275 (P275), or day 2 postpartum (PP). Jugular blood samples were collected at 10-min intervals for 8 hr from all cows. Within 2 hr after completion of blood sampling, animals were slaughtered and the pituitary gland frozen at −196 C. LH pulse frequency/8 hr was reduced (P<.05) during gestation (.5, .2, and 1.5 ± .5/8 hr, for P120, P200, and P275, respectively) and PP (.5 ± .5/8 hr) compared to CY (7.8 ± .5/8 hr). Frequency of LH pulses/8 hr was not different (P>.1) among P120, P200 or PP periods but was different (P<.05) between P200 and P275. There were no differences in LH pulse height (P>.1) among periods; however, pulse amplitude was greatest (P<.05) at P120 (1.3 ± .2 ng/ml) and lowest between P200 and PP (.6 to .8 ± .2 ng/ml). Baseline concentrations of plasma LH did not differ (P>.1) among P and PP periods (.3 ± .1 ng/ml), but were lower (P<.05) than in CY animals (.7 ± .1 ng/ml). Concentration of adenohypophyseal LHRH receptors was approximately two-fold greater (P<.05) at P120 (25.85 ± 2.2 fmol/mg) than at all other periods (9.5 to 14.9 ± 2.2 fmol/mg). Pituitary content of LH was greatest at P120 (1.56 ± .11 ug/mg) and lowest (P<.05) at P275 and PP (0.46 to 0.52 ± .11 ug/mg). Pituitary content of FSH was greatest (P<.05) in P (12.7 to 17.0 ± 1.4 ug/mg) and PP (18.3 ± 1.4 ug/mg) vs CY (5.0 ± 1.4 ug/mg) cows and increased from P120 to PP (P<.05). Results indicate that physiological changes occurring during gestation may have an effect on subsequent function of the adenohypophysis in beef cows.     

Production of estradiol by each ovary during the estrous cycle of cows

The objective of our experiment was to examine changes in serum concentrations of estradiol in each utero-ovarian vein before, during and after gonadotropin surges. Four cows were given prostaglandin F2 alpha (PGF2 alpha) during diestrus and three cows were allowed to cycle spontaneously. All cows had a cannula in each utero-ovarian vein and in one jugular vein. Most cows had two transient rises in estradiol, primarily coming from a single ovary, preceding and after luteinizing hormone (LH) surges. The first rise in estradiol began after luteal regression and was sustained from 48 h before a pre-ovulatory LH surge to the end of the LH surge. The second rise in estradiol was sustained from 72 to 168 h after the end of an LH surge. To determine how rapidly asymmetrical production of estradiol began during luteolysis, several cows were injected with PGF2 alpha during the luteal phase. Blood samples were taken from a jugular and both utero-ovarian veins at hourly intervals before and after PGF2 alpha. Asymmetrical production of estradiol began within 3 h after an injection of PGF2 alpha. We concluded: (1) that a single ovary was responsible for the sustained increases in concentration of estradiol that occur during proestrus to estrus and early diestrus in cows and (2) that cows may have at least one follicle capable of producing estradiol during most days of an estrous cycle, thus little delay in selection of which follicle eventually ovulates occurs after luteal regression.     

Effect of dietary energy, body condition and calf removal on pituitary gonadotropins, gonadotropin-releasing hormone (GnRH) and hypothalamic opioids in beef cows

Beef cows (n = 64) were slaughtered to evaluate effects of dietary energy and calf removal (CR) on hypothalamic and adenohypophysial endocrine characteristics. From d 190 of gestation until parturition, cows received maintenance (ME; n = 32) or low (LE; n = 32) energy diets (ME = 100%, LE = 70% NRC recommendations). After parturition, half (n = 16) of each prepartum diet group received low (LE; n = 32) or high (HE = 130% NRC; n = 32) energy diets. At 30 d postpartum, cows were slaughtered 0 or 48 hr after CR. Hypothalami [preoptic area (POA), hypothalamus (HYP), stalk-median eminence (SME)] and pituitaries were collected. Basal and K(+)-induced release of GnRH from SME, and pituitary luteinizing hormone (LH) and follicle stimulating hormone (FSH) did not differ among groups (P greater than .05). Hypophyseal LH was correlated (P less than .01) with body condition score (BCS) at parturition and slaughter (r = .36 and .47, respectively). Prepartum LE diet increased (P less than .05) met-enkephalin in POA compared to prepartum ME (.59 +/- .05 vs. .44 +/- .04 pmol/mg) regardless of postpartum diet or suckling status. Concentrations of beta-endorphin in combined HYP + POA were decreased (P less than .05) by 48 hr CR (15.1 +/- 1.1 vs. 18.1 +/- 0.7 fmol/mg).(ABSTRACT TRUNCATED AT 250 WORDS)     

Anterior pituitary concentrations of gonadotropins, GnRH-receptors and ovarian characteristics following early weaning in beef cows

Endocrine changes in the hypophyseal-ovarian axis associated with early calf removal were investigated in anestrous beef cows. Tissues were collected and analyzed from multiparous beef cows slaughtered at O (n = 8), 36 (n = 8) or 72 h (n = 8) after calf removal during the fifth week after calving. Cows that exhibited estrus; had postmortem signs of a recent ovulation or had serum concentrations of luteinizing hormone (LH) indicative of an ovulatory surge, were excluded from the analysis. Five control cows that were not slaughtered exhibited estrus from 30 to 84 h after calf removal. Seven additional cows were continuously kept with their calves and did not exhibit estrus until 72 +/- 9 d after calving. Serum concentrations of estradiol-17 beta (estradiol) averaged 8.2 +/- 1.7, 7.5 +/- 2.0 and 9.1 +/- 1.5 pg/ml at 0, 36 and 72 h, respectively, but they averaged 22.8 +/- 4.7 pg/ml prior to estrus in control cows. Therefore, observations were assumed to represent events that occur prior to the rise in serum concentrations of estradiol that occurs during proestrus. Volume of fluid from the largest ovarian follicle tended to be greater (P less than .10) at 72 h (1.5 +/- .2 ml) than at 0 h (1.1 +/- .1 ml) or 36 h (1.0 +/- .1 ml). Follicular-fluid concentrations of estradiol, but not progesterone, were positively correlated (P less than .01) with follicular volume. However, numbers of small (less than 100 microliters), medium (100 to 400 microliters) and large follicles (greater than 400 microliters) as based on fluid volume, as well as follicular-fluid concentrations of estradiol and progesterone, did not differ among treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pituitary responsiveness to GnRH, hypothalamic content of GnRH and pituitary LH and FSH concentrations immediately preceding puberty in gilts

To determine whether pituitary concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH) or hypothalamic content of gonadotropin releasing hormone (GnRH) change before puberty, 40 prepubertal gilts averaging 7 mo of age were slaughtered before or on the second, third or fourth day after relocation and boar exposure. Some gilts responded to relocation and boar exposure as indicated by swollen vulvae, turgid uteri and enlarged ovarian follicles at the time of slaughter. Pituitary concentrations of LH and FSH and hypothalamic content of GnRH were similar between gilts that responded to relocation and boar exposure and gilts that did not respond. In addition, boar exposure and relocation had no effect on pituitary concentrations of LH and FSH or on hypothalamic content of GnRH. To determine whether pituitary responsiveness to GnRH changes before puberty, a third experiment was conducted in which 72 gilts were injected with 400 micrograms of GnRH either before or on the second, third or fourth day after relocation and boar exposure. In gilts that subsequently responded (i.e., ovulated) as a result of relocation and boar exposure, pituitary responsiveness to GnRH was reduced as compared with gilts that failed to ovulate after relocation and boar exposure. Peak concentrations of serum LH after GnRH injection were 4.6 +/- 1.3 vs 9.8 +/- .8 ng/ml for responders vs nonresponders. Peak serum FSH after GnRH injection was also lower for responders than for nonresponders (29.5 +/- 4.2 vs 41.2 +/- 2.4 ng/ml). When compared with controls, relocation and boar exposure did not significantly affect GnRH-induced release of LH and FSH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes and interrelationships among luteal LH receptors, adenylate cyclase activity and phosphodiesterase activity during the bovine estrous cycle

Changes and interrelationships among the cellular mechanisms that may regulate luteal progesterone synthesis during the bovine estrous cycle were studied. Corpora lutea (CL) were enucleated from 30 cows via a transvaginal incision on d 4, 7, 10, 13, 16 and 19 following estrus (estrus = d 0). Mean corpus luteum weight, and luteal progesterone and plasma progesterone concentrations increased (P less than .05) from d 4 to 7 or 10, and declined following luteal regression (d 19). Unoccupied LH receptor (UOR) concentrations increased (P less than .05) more than fourfold from d 4 (38 fmol/mg protein) to d 16 (173 fmol/mg protein) before declining (P less than .05) by d 19 (30 fmol/mg protein). The dissociation constant for UOR concentrations increased (P less than .05) during the estrous cycle. Concentrations of occupied LH receptors (OR) were less (P less than .05) on d 7 than other days; however, the total number of OR/CL increased fourfold from d 4 (47 fmol/CL) to d 10 (221 fmol/CL) and remained similar thereafter. Basal adenylate cyclase, LH-activated adenylate cyclase, and Gpp(NH)p-activated adenylate cyclase activities were greatest (P less than .05) on d 7 to 16 as compared with d 4 and 19. Luteinizing hormone stimulated (P less than .05) adenylate cyclase activity relative to basal activity on d 7, 10, 13, and 16; whereas, Gpp(NH)p stimulated (P less than .05) adenylate cyclase activity relative to basal activity at each period. Phosphodiesterase was 46% greater on d 19 as compared with d 4.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cytoplasmic estrogen and progesterone receptors in canine endometrium during the estrous cycle

Estradiol and progesterone receptors (ER, PR) were characterized and measured in cytosols from canine endometrium, using saturation and sucrose-gradient centrifugation radioassays. Both receptors were demonstrated to be steroid- and tissue-specific saturable proteins, which bound the respective steroids with high affinity (dissociation constant [Kd] approximately 10(-9)M). Serum estradiol, progesterone, and endometrial cytosol receptor concentrations and receptor-binding affinity were measured for 25 bitches from which samples were obtained at 5 stages of the estrous cycle (5 bitches each): anestrus (A), the 3rd day of proestrus (P3), the 3rd day of estrus (E3), the 12th day after onset of estrus (E12), and the 28th day after onset of estrus (E28). Mean (+/- SEM) serum estradiol concentrations were 17.0 +/- 2.2 (A), 55.4 +/- 5.0 (P3), 89.4 +/- 24.9 (E3), 41.0 +/- 5.9 (E12), and 50.6 +/- 3.9 (E28) pg/ml. Mean (+/- SEM) serum progesterone concentrations were 0.4 +/- 0.1 (A), 1.5 +/- 0.2 (P3), 17.3 +/- 7.5 (E3), 41.6 +/- 9.5 (E12), and 25.8 +/- 3.2 (E28) ng/ml. Concentrations of ER increased significantly from 1.06 pmol/g of uterus during stage A to a peak concentration of 6.18 pmol/g of uterus at E12, followed by a gradual decrease to 0.69 pmol/g of uterus by E28. The PR concentrations increased from 3.01 pmol/g of uterus in stage A to 17.32 pmol/g of uterus at P3; PR concentrations, thereafter, decreased gradually to 1.85 pmol/g of uterus by E28. Dissociation constants were significantly higher at E12 for the ER (Kd = 2.6645 X 10(-9)M) and at P3 for the PR (Kd = 5.8282 X 10(-9)M) than at the other stages examined, indicating a decrease in receptor affinity during the periods of high receptor concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)     

Submicroscopic changes in the glandular epithelium of the endometrium in cows during the estrous cycle

During the sexual cycle the structural changes in the cells of the glandular epithelium in the endometrium of a cow are more pronounced than those in the luminal epithelium. Microvilli appear on cell surface during pro-oestrus; in later stages the microvilli disappear and the apical parts penetrate into the lumen. The relative volume of mitochondria increases from pro-oestrus to culminate in oestrus and metoestrus and to drop in dioestrus. The values of the volume and surface area of granular endoplasmic reticulum are also at their highest level in oestrus and metoestrus. The lowest volume and surface area of smooth membranes are recorded in dioestrus, then it markedly increases to culminate in metoestrus. The incidence of lysosomes is variable and their volume culminates in metoestrus. The proliferation of the glandular epithelium has the highest intensity in oestrus and reaches its peak values in metoestrus when the apical parts of cytoplasm contain the largest amounts of smooth-walled vesicles and vacuoles, partly also secretory granules, and when the signs of secretion are observed.     

Plasma gonadotropins and ovarian hormones during the estrous cycle in high compared to low ovulation rate gilts

Mature gilts classified by low (12 to 16 corpora lutea [CL], n = 6) or high (17 to 26 CL, n = 5) ovulation rate (OR) were compared for plasma follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone, estradiol-17beta, and inhibin during an estrous cycle. Gilts were checked for estrus at 8-h intervals beginning on d 18. Blood samples were collected at 8-h intervals beginning on d 18 of the third estrous cycle and continued for one complete estrous cycle. Analysis for FSH and LH was performed on samples collected at 8-h intervals and for ovarian hormones on samples collected at 24-h intervals. The data were standardized to the peak of LH at fourth (d 0) and fifth estrus for the follicular phase and analyzed in discrete periods during the periovulatory (-1, 0, +1 d relative to LH peak), early-luteal (d 1 to 5), mid-luteal (d 6 to 10), late-luteal (11 to 15), periluteolytic (-1, 0, +1 d relative to progesterone decline), and follicular (5 d prior to fifth estrus) phases of the estrous cycle. The number of CL during the sampling estrous cycle was greater (P < 0.005) for the high vs low OR gilts (18.8 vs 14.3) and again (P < 0.001) in the cycle subsequent to hormone measurement (20.9 vs 14.7). For high-OR gilts, FSH was greater during the ovulatory period (P = 0.002), the mid- (P < 0.05) and late-luteal phases (P = 0.01), and tended to be elevated during the early-luteal (P = 0.06), but not the luteolytic or follicular periods. LH was greater in high-OR gilts during the ovulatory period (P < 0.005), but not at other periods during the cycle. In high-OR gilts, progesterone was greater in the mid, late, and ovulatory phases (P < 0.005), but not in the follicular, ovulatory, and early-luteal phases. Concentrations of estradiol-17beta were not different between OR groups during the cycle. Inhibin was greater for the high OR group (P < 0.005) during the early, mid, late, luteolytic, and follicular phases (P < 0.001). The duration of the follicular phase (from last baseline estrogen value to the LH peak) was 6.5 +/- 0.5 d and was not affected by OR group. These results indicate that elevated concentrations of both FSH and LH are associated with increased ovulation rate during the ovulatory phase, but that only elevated FSH during much of the luteal phase is associated with increased ovulation rate. Of the ovarian hormones, both inhibin and progesterone are highly related to greater ovulation rates. These findings could aid in understanding how ovulation rate is controlled in pigs.     

Changes of receptor mRNAs for oxytocin and estrogen during the estrous cycle in rat uterus

Oxytocin receptor (OTR) mRNA levels increase dramatically near term and is potently stimulated by estrogen because increased OTR mRNA levels result from estrogen treatment in ovariectomized rat uterus. In this study, OTR, estrogen receptor (ER) alpha and ERbeta mRNA levels in the rat uterus during the estrous cycle were examined by quantitative RT-PCR. OTR mRNA levels during the estrous cycle began to increase on diestrus (P<0.05, vs value on estrus), reached maximal increase both in the morning (1000-1130 hr) and afternoon (1600-1630 hr) on proestrus (P<0.01, vs metestrus, diestrus and estrus) and then declined on estrus. In contrast ER alpha mRNA levels began to decrease on diestrus, reached statistical significance both in the morning and the afternoon on proestrus (P<0.01, vs metestrus, diestrus and estrus) and returned to the value of metestrus on estrus. ERbeta mRNA levels were low in the morning and the afternoon on proestrus (P<0.01, vs metestrus and estrus) and also returned to metestrus values on estrus. Treatments with estrogen for 3 days significantly decreased both ERalpha and ERbeta mRNA levels. It can be concluded from these results that during the estrous cycle, OTR mRNA levels in rat uterus predominantly increase at proestrus with a decrease in ERalpha and ERbeta mRNA levels, which is probably due to the increased estrogen levels in circulation before ovulation.     

Endocrine changes and ultrasonography of ovaries in suckled beef cows during resumption of postpartum estrous cycles

Changes in follicular and luteal structures were assessed and concentrations of estradiol and progesterone were measured in 13 Hereford X Angus suckled beef cows during resumption of estrous cycles. Transrectal ultrasonography was used to monitor follicular size, ovulation, and formation and regression of the corpus luteum (CL). The interval from parturition to first postpartum ovulation (FO) was 82 +/- 4.7 d. Serum progesterone remained low before FO. One cow exhibited standing estrus, two cows showed other signs of estrus, and 10 displayed no signs of behavioral estrus preceding FO. All cows exhibited standing estrus before the second postpartum ovulation (SO). All cows had a short luteal phase after FO, with an average interval of 8.5 +/- .2 d between FO and SO. Concentrations of estradiol in serum during the 8 d preceding ovulation were similar before FO and SO. Maximal diameter of the preovulatory follicle was similar before FO and SO. However, the ovulatory follicle was larger in diameter at 2 d (P = .02) and 3 to 8 d (P less than .005) before FO than before SO. The time from detection until ovulation was less (P = .005) for the ovulatory follicle preceding SO than for the follicle associated with FO (8.5 vs 10.2 d, respectively, SE = .4). The second-largest follicle was larger (P less than .005) in diameter during the 8 d preceding the FO than before the SO. The difference in size between the ovulatory follicle and the second-largest follicle on the day before ovulation was greater (P less than .005) preceding SO than preceding FO (8.7 vs 6.6 mm, respectively, SE = .4).(ABSTRACT TRUNCATED AT 250 WORDS)     

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.