Western immunoblotting in avian shell gland sample immunoblotting methods.

Avian shell gland tissue was subjected to Western blot analysis using anti-human estrogen receptor-alpha antibody H222. Initial attempts to obtain consistent, high-quality blots were unsuccessful because, as it turned out, excessive lipid in tissue preparations interfered with protein separation. Incremental additions of acetone eventually proved to be the critical step in solubilizing lipids and allowing consistent separation of bands on gels. A detailed description of the methodology is presented.

Estrogen receptor-alpha populations change with age in commercial laying hens.

Older hens in production lay larger but fewer eggs than younger birds, and the incidence of soft and broken shells is greater in older hens than younger. These changes are attributable at least in part to changing hormone profiles and diminished ability of the hen to transport calcium at the duodenum. In further exploration of this relationship, a study was conducted with three ages of Hy-Line W-36 birds: prelay pullets (PL; 19 wk, 0% production), peak-production hens (PP; 29 wk, approximately 93% production), and late-stage hens (LS; 71 wk, approximately 80% production). Hens from the PP and LS groups were palpated for presence of an egg in the shell gland; hens were then euthanized and tissues (kidney, shell gland, hypothalamus) were removed for quantification of estrogen receptor-alpha (ERalpha) populations via immunocytochemical and Western blot analyses. Localization of ERalpha by immunostaining in the shell gland showed differences among age groups; however, no differences were noted in localization of ERalpha between age groups in the kidney and hypothalamus. In both the kidney and the shell gland there was a decrease in the amount of ERalpha, as detected by immunoblotting, in the LS hens compared to PL and PP birds (P < 0.05). The results suggest that failure of calcium regulating mechanisms with age may be mediated at least in part through the reduced populations of estrogen receptors in certain critical tissues.

Effect of glucose availability on pulsatile luteinizing hormone release in rams before and after castration.

The hypothesis tested was that availability of glucose modulates the control of luteinizing hormone (LH) release. A second objective was to determine the role of testicular hormones in the control of pulsatile LH secretion during depressed blood glucose. Serial blood samples were collected at 15 min intervals for 8 h from intact pubertal Suffolk rams (n = 8; 7 months old) on consecutive days (Days 1, 2 and 3). Rams were castrated after sampling on Day 3 and samples were collected 3 weeks later on consecutive days (Days 4, 5 and 6). Insulin (120 units, iv) was given at Hour 4 of each of the six days to lower blood glucose. On Days 1 and 4, no other treatments were given (Control). On Days 2 and 5, LH releasing hormone (LHRH; 5 ng/kg, iv) was given at Hours 5, 6 and 7 to assess the ability of the pituitary to release LH. On Days 3 and 6, N-methyl-D,L-aspartate (NMA; 5 mg/kg, iv) was given at Hours 5, 6 and 7 to assess the ability of the hypothalamus to release LHRH. Insulin reduced plasma glucose by 52% for at least 3 h (P < 0.001). Insulin reduced the mean LH concentration (P < 0.05) and tended to reduce the LH response area (P < 0.10) in castrated animals during the control period. LHRH increased LH pulse number (P < 0.001) in intact rams and increased mean LH concentration (P < 0.01), LH pulse amplitude (P < 0.05) and LH response area (P < 0.01) in castrated animals compared to respective control periods. NMA increased mean LH concentration in intact rams (P < 0.0001) but did not affect mean LH in castrates. NMA increased LH pulse number in rams (P < 0.0001) but decreased number of pulses in castrates (P < 0.0001) compared to control periods. NMA increased LH pulse amplitude in both intact (P < 0.001) and castrated animals (P < 0.05). In conclusion, these results support the hypothesis that blood glucose concentrations influence the control of LH release in sheep. In addition, LH release in response to the LHRH secretagogue, NMA, is positively influenced by testicular hormones.

Concentrations of gonadotropins, estradiol and progesterone in sows selected on an index of ovulation rate and embryo survival.

The objective of this study was to determine concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P4) and 17beta-estradiol (E2) in sows from a line selected on an index which emphasized ovulation rate (Select) and from a control line. A further classification of the sows in each line was made according to the estimated number of ovulations during an estrous cycle. Sows in the Select line were ranked into a high (HI) or low group (LI) when their estimated number of ovulations were 25 or more and 14 to 15, respectively. Sows of the control line were classified into groups as high (HC) or low (LC) when the estimated values for ovulation rate were 14-15 and 8-9 ovulations, respectively. Blood samples were collected every 12 h during a complete estrous cycle and samples were analyzed for concentrations of FSH and LH. Samples collected every 24 h were assayed for P4 and E2. Mean concentrations of FSH, LH, P4 and E2 did not differ (P > 0.10) between lines or between HI and LI or HC and LC groups. Selection of pigs for ovulation rate and embryonal survival did not affect concentrations of FSH, LH, P4 and E2 in sows during the estrous cycle.

Gonadotropin secretion and development of ovarian follicles during oestrous cycles in heifers treated with luteinizing hormone releasing hormone antagonist.

The hypothesis tested was that reduced LHRH stimulation of the anterior pituitary would lead to attenuated development of ovarian follicles as a result of reduced gonadotropin secretion during oestrous cycles of cattle. Twenty heifers were randomly assigned to be treated ( n = 5/treatment) with an antagonist to LHRH (LHRH-Ant) 1) from Day 2 to 7 (Day 0 = behavioural oestrus), 2) Day 7 to 12, 3) Day 12 to 17, 4) or serve as untreated control animals. LHRH-Ant suppressed LH pulses of heifers in all treatment groups from treatment initiation through Day 17 as compared with untreated control heifers [Peters et al., 1994. Luteinizing hormone has a role in development of fully functional corpora lutea (CL) but is not required to maintain CL function in heifers. Biol. Reprod., 51 (1994) 1248-1254]. Circulating concentration of FSH from Day 8 to 10 of the oestrous cycle did not increase in heifers treated with LHRH-Ant from Day 2 to 7 or Day 7 to 12; however, there was increased (P < 0.05) FSH from Day 8 to 10 of the oestrous cycle in heifers treated with LHRH-Ant from Day 12 to 17 and control heifers. Compared with control heifers, heifers treated with LHRH-Ant from the Day 2 to 7 had suppressed (P < 0.05) size and persistence of the first and second dominant ovarian follicles. Heifers treated with LHRH-Ant from Day 7 to 12 had suppressed size (P < 0.05 and tended (P < 0.10) to have a shorter persistence of the second dominant ovarian follicle compared with control heifers. Heifers treated with LHRH-Ant from Day 12 to 17 had a similar (P > 0.10) size and persistence of dominant ovarian follicles but had reduced (P < 0.10) numbers of large follicles compared with control heifers. Heifers treated with LHRH-Ant from Day 2 to 7 had lower (P < 0.01) concentrations of 17 beta-oestradiol during the treatment period and tended (P < 0.10) to have lower concentrations of 17 beta-oestradiol from Day 7 to 12 of the oestrous cycle compared with control heifers. Heifers treated with LHRH-Ant from Day 7 to 12 or Day 12 to 17 had similar (P > 0.10) circulating LH concentrations of l7 beta-oestradiol compared with control heifers. Reduced LHRH stimulation of the pituitary from Day 2 to 12 of the oestrous cycle and the resulting modulation in circulating LH and FSH led to suppressed ovarian follicular development and oestradiol secretion. After Day 12 of the oestrous cycle, reduced LHRH stimulation of the anterior pituitary did not lead to altered ovarian follicular development to the extent as reduced LHRH stimulation before Day 12 although pulsatile release of LH was similarly suppressed by treatment with the LHRH-Ant.

Comparison of circulating concentrations of reproductive hormones in boars of lines selected for size of testes or number of ovulations and embryonal survival to concentrations in respective control lines.

The objectives of this study were to determine whether circulating concentrations of genadotropins and gonadal hormones of boars were altered as a result of selection of pigs for size of testes or for embryonal survival and(or) number of ovulations. Included in Exp. 1 and 2 were boars with the greatest estimated paired weight of testes (TS) and boars from a control (C) line. Concentrations of FSH were similar (P > .10) in boars from the TS and C lines. In Exp. 3, 4, and 5, circulating concentrations of FSH and 17 beta-estradiol (E2) were evaluated in neonates, during pubertal development, and in mature boars of lines selected for an index of number of ovulations and embryonal survival (I), and data were compared to those for boars from a respective C line. Concentrations of E2 were not different in boars from the I line and those from the C line during the early neonatal period but were greater (P < .05) in boars of the C line than in those from the I line during pubertal development. Concentrations of FSH were greater (P < .05) in mature boars from the I line than in those from the C line. In summary, selection for size of testes did not influence circulating concentrations of FSH in mature boars. The secretory pattern of E2 in boars before puberty changed as a result of selection for embryonal survival and number of ovulations in females of the I line, and the different patterns of circulating E2 early in life may result in enhanced circulating concentrations of FSH in adult boars of the I line compared with boars of the C line.

Uterine mass and uterine blood volume in mice selected 21 generations for alternative criteria to increase litter size.

Lines of mice, selected for 21 generations using alternative criteria to increase litter size, were evaluated for uterine mass and uterine blood volume to help explain differences in uterine capacity. For this study, mice were sampled from Generation 27, the sixth generation after relaxation of selection. Mice came from all four criteria of selection (LS = selection on number born to unaltered females; IX = selection on index of ovulation rate and ova success; UT = selection on uterine capacity; and LC = unselected control) in each of three replicates (a total of 12 lines). Measurement was at one of two stages, either 3 d or 6 d of gestation. Matings were at 10 wk of age, and a total of 508 mice (17 to 26 per line-day of pregnancy subclass) were measured. The mean of the three selected groups exceeded the control in uterine mass (P < .001), uterine blood volume (P < .002), uterine mass/body mass (P < .03), and uterine blood volume/body mass (P < .04) but not in uterine blood volume/uterine mass. Greater uterine mass and concomitantly greater uterine blood volume may have been partly responsible for greater uterine capacity resulting from LS, IX, and UT selections.

Effects of 17 beta-estradiol on distribution of pituitary isoforms of luteinizing hormone and follicle-stimulating hormone during the follicular phase of the bovine estrous cycle.

The objective of this study was to examine the influence of 17 beta-estradiol (E2) on distribution of LH and FSH isoforms during the follicular phase of the bovine estrous cycle prior to the preovulatory surges of LH and FSH. On Day 16 of the estrous cycle (Day 0 = estrus), intact controls (CONT; n = 4) were treated with prostaglandin F2 alpha (PGF2 alpha) to induce luteal regression and initiation of the follicular phase. Other cows were also treated with PGF2 alpha and either ovariectomized (OVX; n = 5) or ovariectomized and given E2 implants (OVXE; n = 6) to mimic the pattern of increasing E2 concentrations during the follicular phase of the estrous cycle. Pituitaries were collected 40 h after treatment with PGF 2 alpha or ovariectomy (0 h). Aliquots of pituitary extracts were chromatofocused on pH 10.5-4.0 gradients. The LH resolved into thirteen isoforms (designated A-L and S, beginning with the most basic form) while FSH resolved into nine isoforms (designated I-IX, beginning with the most basic form). The percentage of LH as isoform F (elution pH = 9.32 +/- 0.01) was greater (p < 0.05) in the OVX group (48.5%) than in the OVXE group (45.0%). LH isoforms I (elution pH = 6.98 +/- 0.01) and J (elution pH = 6.48 +/- 0.01) were more abundant (p < 0.05) in cows from the OVXE (2.3 and 5.8%, respectively) than the OVX group (1.4 and 3.7%, respectively). Distribution of LH isoforms in cows from the three groups did not differ (p > 0.10).(ABSTRACT TRUNCATED AT 250 WORDS)

Melengestrol acetate at greater doses than typically used for estrous synchrony in bovine females does not mimic endogenous progesterone in regulation of secretion of luteinizing hormone and 17 beta-estradiol.

Our working hypothesis was that doses of melengestrol acetate (MGA) greater than those typically administered in estrous synchrony regimens would regulate secretion of LH and 17 beta-estradiol (E2) as endogenous progesterone (P4) does during the midluteal phase of the estrous cycle. We also hypothesized that endogenous P4 from the CL would interact with MGA to further decrease the frequency of LH pulses and E2. Cows on Day 5 of their estrous cycle (Day 0 = estrus) were randomly assigned to an untreated control group (CONT, n = 5) or to one of six MGA treatment groups (n = 5 per group): 1) MGA administered orally each day via a gelatin capsule at a dose of 0.5 mg MGA/cow with the CL present (0.5CIL); 2) 0.5 mg MGA/cow daily in the absence of CL (0.5NO); 3) 1.0 mg MGA with CL present (1.0CL); 4) 1.0 mg MGA without CL (1.0NO); 5) 1.5 mg MGA with CL present (1.5CL); 6) 1.5 mg without CL (1.5NO). MGA was administered for 10 days (Day 5 = initiation of treatment). To regress CL, cows assigned to groups without CL received injections of prostaglandin F 2 alpha (PGF 2 alpha; 25 mg) on Days 6 and 7 of their estrous cycle. All cows were administered PGF2 alpha at the end of the 10-day treatment period. During the treatment period, daily blood samples were collected to determine concentrations of E2. Serial blood samples were collected at 15-min intervals for 24 h on Days 8, 11, and 14 to determine pattern of LH secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Dosage of the synthetic progestin, norgestomet, influences luteinizing hormone pulse frequency and endogenous secretion of 17 beta-estradiol in heifers.

The objective of this study was to determine whether there were doses at which the synthetic progestin, norgestomet, could mimic midluteal phase concentrations of progesterone in regulating the secretion of LH and 17 beta-estradiol in bovine females. Heifers were randomly assigned to one of five groups to receive: 1) one (1Norg, n = 5), 2) two (2Norg, n = 5), 3) four (4Norg, n = 5), or 4) eight (8Norg, n = 5) norgestomet implants or to serve as untreated control heifers (control, n = 5). On Day 7 (Day 0 = behavioral estrus), implants containing norgestomet were inserted, and they remained in place for 10 days. All heifers implanted with norgestomet were treated with 25 mg prostaglandin F2 alpha (PGF2 alpha) on Days 7 and 8 to lyse the CL. Controls were treated with 25 mg PGF2 alpha at the time norgestomet implants were removed from heifers of the other treatment groups. Blood samples were collected every 15 min for 24 h on Days 10 and 16 to determine the frequency of LH pulses. Beginning 24 h after removal of implants, samples of blood were collected at 4-h intervals for 96 h to determine the time of the preovulatory surge of LH. Daily blood samples were collected from Day 2 to Day 48 to determine concentrations of progesterone, and samples collected between Days 2 and 17 were used to determine concentrations of 17 beta-estradiol. Ultrasonography was performed daily from Day 2 until Day 23 to evaluate ovarian follicular development.(ABSTRACT TRUNCATED AT 250 WORDS)

Increasing concentrations of 17 beta-estradiol has differential effects on secretion of luteinizing hormone and follicle-stimulating hormone and amounts of mRNA for gonadotropin subunits during the follicular phase of the bovine estrous cycle.

The hypothesis tested was that 17 beta-estradiol (E2) would increase amounts of mRNA for alpha, LH beta, and FSH beta subunits during the follicular phase of the estrous cycle prior to the preovulatory surge of gonadotropins in cows. On Day 16 (Day 0 = estrus) of the estrous cycle, all cows were treated with prostaglandin F2 alpha (PGF2 alpha). Cows served as intact controls (CONT, n = 4) were ovariectomized (OVX, n = 5), or were ovariectomized and administered E2 (OVXE, n = 6) in increasing doses starting at the time of treatment with PGF2 alpha. Cows were bled for 6 h before and for 40 h after PGF2 alpha treatment to characterize pulsatile secretion of LH and FSH. Forty hours after PGF2 alpha treatment, pituitaries were collected for evaluation of amounts of mRNA for alpha, LH beta, and FSH beta subunits. Amplitude of LH pulses was greater (p < 0.05) in cows from the OVXE than from the CONT group. Concentrations of FSH were greater in cows from both the OVXE and OVX (p < 0.01) groups than from the CONT group. Amounts of mRNA for alpha and FSH beta subunits were greater (p < 0.01) in pituitaries of cows from the OVX than from the CONT or OVXE groups. Amounts of mRNA for LH beta subunit in pituitaries of cows from the OVX group tended to be greater (p < 0.08) than from the CONT group. Cows in the OVXE group tended (p < 0.08) to have greater amounts of mRNA for FSH beta subunit than did CONT cows.(ABSTRACT TRUNCATED AT 250 WORDS)

Luteinizing hormone has a role in development of fully functional corpora lutea (CL) but is not required to maintain CL function in heifers.

We tested the hypothesis that endogenous pulses of LH have a role in development and maintenance of CL during the estrous cycle of the bovine female. Twenty heifers were synchronized to estrus by treating two times with prostaglandin F2 alpha 11 days apart (Day 0 = behavioral estrus). Heifers were then randomly assigned to one of four treatments (n = 5/group). Heifers were treated with an antagonist to LHRH (LHRH-Ant; N-Ac-D-Nal[2]1,4Cl-D-Phe2,D-Pal[3]3,D-Cit6,D-Ala10- LHR H; 10 micrograms/kg body weight) or vehicle (5% mannitol) once every 24 h: 1) LHRH-Ant Days 2-7, 2) LHRH-Ant Days 7-12, 3) LHRH-Ant Days 12-17, 4) no LHRH-Ant (control). Blood samples were collected from the jugular vein twice daily on Days 0-24, and area under the profile of progesterone in circulation during the luteal phase of the estrous cycle was characterized from the start of each treatment period until the demise of CL or Day 24, whichever came first. Luteolysis was considered to have occurred when three consecutive samples contained less than 1 ng progesterone/ml plasma. Areas under the profile of progesterone in circulation during the luteal phase of the estrous cycle were compared to those of heifers from the control group for the same period. LHRH-Ant treatment diminished LH pulses in all treatment groups compared to control (p < 0.05). Treatment with LHRH-Ant on Days 2-7 diminished function of CL (3.72 +/- 0.93 vs. 7.36 +/- 1.02 units, respectively; p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Serum cholesterol concentration of mice selected for litter size and its relationship to litter size and testis mass.

This study assessed the genetic relationship between litter size and serum cholesterol concentration and between litter size and testis mass in mice. Mice were from a long-term experiment in which selection had occurred for 21 generations in three replicated lines per criterion of selection (LS = selection to increase litter size based on number born; LC = unselected control). Thereafter, random mating within lines was practiced. Serum cholesterol concentrations were evaluated in female and male mice from two replicates at Generation 29 and one replicate at Generation 30. Body weights and blood samples were collected from primiparous females 8 d after weaning their pups. Data from males were collected as they came out of breeding cages. In addition, the testes were excised, stripped clean of connective tissue and the epididymides, and weighed. Means for body mass of females and males, serum cholesterol, number born, and testis mass were as follows: 35.2 vs 32.5 g (P < .09), 33.9 vs 30.7 g (P < .08), 117.5 vs 110.5 mg/dL (P < .08), 14.0 vs 10.3 pups (P < .04), and 126 vs 122 mg, respectively, for LS and LC. Serum cholesterol was greater in males than in females (133.3 vs 95.1 mg/dL; P < .001), but there was no interaction between sex and selection criterion. Serum cholesterol concentration was not correlated phenotypically to number born or body mass, but it had a small negative relationship with testis mass. Therefore, we concluded that selection for litter size tended to increase serum cholesterol in addition to the increase in number born but did not change testis mass.

The effect of protein intake on boar libido, semen characteristics, and plasma hormone concentrations.

To determine the effect of low protein intake on boar libido, semen characteristics, and plasma hormone concentrations, 20 crossbred boars (1 yr of age) were divided into 10 littermate pairs, and boars from within pairs were fed 44 g/kg of BW.75 per day of either a low-protein diet (7% CP) or a control diet (16% CP) with the same energy content (3.41 Mcal of ME/kg). During the first 16 wk and from wk 19 to 23, semen was collected two times per week. During wk 17 and 18, boars were subdivided within dietary treatment and semen was collected either two or seven times per week. Blood samples were collected at 12-min intervals for 6 h before and 1 h after an intravenous injection of GnRH (375 ng/kg of BW) during wk 24. All plasma samples were analyzed for LH and pooled samples were analyzed for estradiol-17 beta and testosterone. Boars with low protein intakes required more time to start ejaculation (P = .11, wk 0 through 7; P < .04, after wk 7), had a shorter duration of ejaculation (P < .09, wk 19 through 23), and had reduced semen volumes (P < .01, after wk 7) compared with boars on the control treatment. There was no interaction between dietary treatment and semen collection frequency (P > .39) for any of the semen or libido measurements. Testosterone and LH concentrations were not affected by protein intake (P > .5). However, concentration of estradiol-17 beta was greater in boars fed the control diet than in boars fed the low-protein diet (582 vs 202 pg/mL, respectively; P < .08). Estrogen concentrations in boars were negatively correlated with the time required for the boar to start ejaculating (r2 = .72). Boars with low protein intakes had reduced libido and semen volume. This reduction in libido and semen volume may be a result of a decrease in estradiol-17 beta concentration in circulation.

The effect of energy and protein intakes on boar libido, semen characteristics, and plasma hormone concentrations.

To determine the effect of energy and protein intakes on boar libido, semen characteristics, and plasma hormone concentrations, 24 crossbred boars (1 yr of age) were allotted to one of three dietary treatments. Two protein levels (7.7 and 18.1 g/d of lysine) and two energy levels (6.1 and 7.7 Mcal/d of ME) were combined to create 1) low-energy and low-protein (ep), 2) low-energy and high-protein (eP), and 3) high-energy and high-protein (EP) treatments. Semen was collected two times per week for 27 wk. During wk 28, blood samples were collected before and after a intravenous injection of GnRH (375 ng/kg of BW). The EP boars had higher (P < .01) ADG than the eP boars (373 vs 169 g), which had higher (P < .01) ADG than the ep boars (169 vs 92 g). Fewer EP and eP boars (zero of eight and two of eight, respectively) than ep boars (five of eight) consistently refused to mount the collection dummy (P < .05). During the final 20 wk of the semen-collection period, EP and eP boars had 33% longer durations of ejaculation (P < .04), had 60% greater semen and gelatinous fraction volumes (P < .03), but had 25% lower sperm concentration in their semen (P < or = .12) than ep boars. The EP boars produced 38% more sperm than the ep boars (P < .10). Baseline LH concentration, LH pulse frequency, LH pulse amplitude, testosterone concentration, and estradiol-17 beta concentrations were not affected by treatment (P > .10).(ABSTRACT TRUNCATED AT 250 WORDS)

Season of the year influences concentration and pattern of gonadotropins and testosterone in circulation of the bovine male.

Serum concentrations of LH vary with season of the year in ovariectomized beef and dairy cows. The objective of the present study was to determine whether concentrations and profile of the gonadotropins and testosterone (T) in circulation vary with season of year in bovine males of a composite breed type (beef). Five INTACT and five gonadectomized (GNX) males that were 22 mo of age at the initiation of the study were used. All of the males utilized were maintained on pasture throughout the year at the latitude of 41 degrees N. Blood samples were collected in a serial regimen (10-min intervals for 24 h) at the spring and fall equinox and the summer and winter solstice. Concentrations of LH were quantified in all samples, and concentrations of T were determined in all samples of the INTACT group. Concentrations of FSH and 17 beta-estradiol (E2) were quantified in pooled samples for all animals, while concentrations of T were determined in pooled samples for males from the GNX group. Pulse frequency of LH or T and mean concentration of FSH did not vary with season of the year. A seasonal effect on mean concentration of LH (p < 0.01) occurred in males of the GNX group, with mean concentrations of LH being greatest during the spring equinox and lowest during the winter solstice. Season had an effect on pulse amplitude of LH (p < 0.01), mean concentration of T (p < 0.01), and pulse amplitude of T (p < 0.05) in males from the INTACT group.(ABSTRACT TRUNCATED AT 250 WORDS)

Progesterone, 17 beta-estradiol, and opioid neuropeptides modulate pattern of luteinizing hormone in circulation of the cow.

The working hypothesis for this study was that 17 beta-estradiol (E2), progesterone (P4), and opioid neuropeptides modulate frequency of pulsatile release of LH during the luteal phase of the bovine estrous cycle. On Day 8 of the estrous cycle (Day 0 = estrus), 20 cows were ovariectomized (OVX) and immediately received one of three steroid replacement treatments: P4 alone (n = 6), E2 alone (n = 7), or P4 and E2 (n = 7). To characterize the pattern of LH in peripheral circulation, serial blood samples were collected (12-min intervals for 22 h) on the fifth day following OVX. LHRH was administered after the 22-h sample was taken. Naloxone (opioid neuropeptide antagonist; 0.5 mg/kg) was administered on the sixth day after OVX, and change of concentration of LH in peripheral circulation was monitored. Pulse frequency of LH and mean concentration of LH were lower (p < 0.01) in cows treated with P4 and E2 compared to cows treated with P4 alone. Moreover, cows treated with E2 alone had a greater (p < 0.01) frequency of LH pulses and mean concentration of LH compared to cows treated with P4 alone. The greatest amplitude (p < 0.01) of LH release after LHRH was in cows treated with E2 alone. The greatest percentage increase (p < 0.01) in LH after administration of naloxone occurred in cows treated with P4 alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Pregnancy rate is greater when the corpus luteum is present during the period of progestin treatment to synchronize time of estrus in cows and heifers.

Our hypothesis was that conception in bovine females would be enhanced if the corpus luteum was present during the period of progestin treatment to synchronize estrus. In this study, 67 heifers (one replicate) and 124 cows (two replicates) were randomly assigned to one of two treatment groups. Seven days after estrus (Day 0), all animals were implanted with norgestomet and the implant remained in place for 10 days. All implants were removed on Day 17. Cows and heifers in one group received prostaglandin F2 alpha (PGF2 alpha) on Day 7 of the estrous cycle (PG 7; norgestomet without corpus luteum), and animals in the second group received PGF2 alpha on Day 17 (day of implant removal; PG 17; norgestomet with corpus luteum). All heifers and cows exhibiting behavioral estrus were artificially inseminated 12 h after estrus was detected during a 7-day period following removal of norgestomet. Blood samples were collected from cows of replicate 1 to determine serum concentrations of progesterone and 17 beta-estradiol. Percentage of females that had calves as a result of artificial insemination was greater (p < 0.01) in the PG 17 group (87% and 78% cows [two replicates] and 58% heifers) compared to the PG 7 group (31% and 44% cows [two replicates] and 41% heifers).(ABSTRACT TRUNCATED AT 250 WORDS)

Increasing exogenous progesterone during synchronization of estrus decreases endogenous 17 beta-estradiol and increases conception in cows.

The objectives of this study were to determine the effects of dose of exogenous progesterone (P4) prior to artificial insemination on concentrations of 17 beta-estradiol (E2) and on conception rates in bovine females. Heifers (n = 100) and cows (n = 100) received P4-releasing intravaginal devices (PRIDs) to produce two different circulating concentrations of P4. All animals received a single PRID 10 days before (Day-10) the start of the breeding season (Day 0). In animals that received the low dose of P4 (1 PRID, target concentration of 2-3 ng/ml of plasma), the original PRID remained in place for 10 days. In animals that received the larger dose of P4 (2 PRIDs, target concentration of 5-8 ng/ml of plasma), an additional PRID was inserted on Day -9. To maintain concentrations of P4 in the 2-PRID group, the PRIDs inserted on Days -10 and -9 were replaced with new PRIDs on Days -5 and -4, respectively. Prostaglandin F2 alpha (25 mg) was administered to all animals on Days -9 and -3 to remove the endogenous source of P4. Following PRID removal, animals were artificially inseminated 12 h after signs of behavioral estrus were observed. A treatment-by-day interaction (p < 0.0001) was observed for concentrations of P4 in circulation of both heifers and cows. Animals that received 2 PRIDs had greater (p < 0.001) concentrations of P4 by Day-8 of treatment than animals that received 1 PRID. In cows that received 1 PRID, concentrations of E2 increased 2.4-fold from Day-10 (6.8 +/- 1.0 pg/ml) to Day-2 (16.7 +/- 1.4 pg/ml).(ABSTRACT TRUNCATED AT 250 WORDS)

Yearling bulls shorten the duration of postpartum anestrus in beef cows to the same extent as do mature bulls.

The objective of this study was to determine whether yearling bulls, when pastured with cows, reduced the duration of postpartum anestrus to the same extent as did mature bulls. This experiment was conducted over a 3-yr period. Cows were stratified by parity group to achieve 37% 2-yr-old and 63% mature (> 2-yr-old) cows within each treatment group (approximately 50 cows per treatment per year). Cows were assigned in the order in which they calved to one of three treatment groups: 1) isolated from bulls (NBE; n = 158); 2) exposed to mature bulls that were > 3 yr of age (MBE; n = 154); or 3) exposed to bulls that were 1 yr of age (YBE; n = 152). Beginning the 2nd wk after calving, cows were pastured with either sterile bulls that were 1 yr (YBE) or > 3 yr of age (MBE) (three bulls per treatment group). Blood samples were collected twice weekly from late March until mid-July each year. Cows with serum concentrations of progesterone > 1 ng/mL for two consecutive sampling periods were assumed to have initiated estrous cycles after calving. Duration of postpartum anestrus in cows exposed to yearling bulls (YBE = 61.8 +/- 1.8 d) did not differ (P > .10) from duration of postpartum anestrus in cows exposed to mature bulls (MBE = 59.5 +/- 1.7 d). Duration of postpartum anestrus was shorter (P < .01) for cows exposed to bulls (MBE+YBE = 61.0 +/- 1.7 d) than for cows isolated from bulls (NBE = 72.3 +/- 1.8 d).(ABSTRACT TRUNCATED AT 250 WORDS)