We hypothesized that increasing concentrations of progesterone (P4) after artificial insemination would increase fertility. Our objective was to assess changes in ovarian structures, incidence of ovulation, and change in serum P4 in response to GnRH, human chorionic gonadotropin (hCG), or exogenous P4 (controlled internal drug release; CIDR insert) treatment beginning 4 to 9 d after artificial insemination (d 0) and again 7 d later (experiment 1). Blood was collected from 753 cows in 3 herds on d 0 and 7. Ovaries of 162 cows were scanned and mapped to confirm the presence of a corpus luteum (CL), and cows were assigned randomly to serve as controls (n = 41) or to receive a CIDR insert for 7 d (n = 41), 100 microg of GnRH (n = 40), or 3,300 IU of hCG (n = 40). More cows were induced to ovulate in response to GnRH (60%) and hCG (78%) compared with controls (2.4%). Compared with controls, cows treated with GnRH or hCG had more induced CL (d 7) and more total CL (d 7), but serum P4 was increased only in response to hCG. Largest follicle diameters on d 7 were less after GnRH and hCG, but total follicular volume on d 7 was reduced by GnRH, hCG, and CIDR, compared with that of controls. Volume of the original luteal structures was increased by hCG but tended to be reduced by CIDR and GnRH compared with luteal volume in controls. Total CL volume was increased by hCG, but reduced by CIDR, compared with CL volume of controls. Conception rates and pregnancy survival were assessed in response to the same treatments described in experiment 1: controls (n = 708), CIDR (n = 711), GnRH (n = 719), and hCG (n = 714). Tendencies for interactions of treatment x herd and treatment x lactation group were detected, but no 3-way interactions were found. Treatment with hCG increased conception rates in second-lactation cows. The CIDR tended to increase, and hCG increased, conception rates in 2 herds, whereas the CIDR decreased conception rates in 1 herd. Pregnancy survival was reduced by GnRH compared with that in controls. We concluded that GnRH and hCG effectively induced ovulation, and increased number of CL, but only increased serum P4 in hCG-treated cows. Further, treatment with the CIDR or hCG increased conception rates but only in some herds.

BackgroundSeveral alternatively-spliced mRNA transcripts of the follicle stimulating hormone receptor (FSHR) have been identified in sheep, including FSHR-1 (G protein-coupled form), FSHR-2 (dominant negative form), and FSHR-3 (growth factor type-1 form). Our objective was to determine which of these variants is predominantly expressed in follicles collected from ewes at various times after estrus.MethodsSuffolk-cross ewes (n = 8) were allowed to come into estrus naturally and were euthanized 24 (n = 3), 36 (n = 3), or 48 (n = 2) hours after the onset of estrus. All visible follicles were measured, aspirated and pooled according to follicular diameter: small (<= 2.0 mm), medium (2.1-4.0 mm), large (4.1-6.0 mm), and preovulatory (> = 6.1 mm). Aspirated cells were separated from follicular fluid by centrifugation. Total RNA was extracted from cell pellets and reverse transcribed. The resulting cDNA was subjected to qPCR, using primer sets designed to amplify each variant specifically. Gene expression was normalized to that of beta–actin within samples, and compared by analysis of variance with the level of significant differences set at p < .05.ResultsRelative expression of FSHR-3 exceeded that of both FSHR-1 and FSHR-2 in medium follicles, and tended to be higher in small follicles (p = .09) regardless of time after onset of estrus, and thus results from different time points were pooled. Expression of FSHR-3 was greater than that of FSHR-2 and luteinizing hormone receptor (LHR) in small and medium follicles. Expression of LHR was greatest in preovulatory follicles.ConclusionsThese experiments show that in addition to the well characterized G protein-coupled form of the FSHR, alternatively spliced variants of the FSHR may participate in follicular dynamics during follicular waves of the sheep estrous cycle. Furthermore, these results indicate that an “alternatively” spliced form of the FSHR (FSHR-3) is the predominant form of the FSHR in the sheep.

Programs for developing replacement heifers are designed for heifers to calve at 2 yr of age and to extend their stayability in the herd and minimize feed cost. The experimental objective was to determine whether developing prepubertal heifers on less dietary energy and to a BW of 55% rather than 65% of mature BW at 14 mo of age would compromise ovarian development and reduce fertility. In a 3-yr study, 8-mo-old Angus (n = 60/yr) and composite MARC II (n = 60/yr) heifers were assigned equally by age, BW, and breed to receive either a low (LG) or high (HG) BW gain diet fed to achieve an ADG of either 0.45 or 0.8 kg/d from 8 to 15 mo of age, including the first 21 d of breeding, and then transferred to pasture. At 14 mo, heifers were housed with fertile bulls for 47 d. Estrus was monitored for 21 d. Within 12 h after detection of estrus, ovarian length and height, preovulatory follicle diam., and antral follicle count (AFC) were measured by transrectal ultrasonography. Corpus luteum (CL) volume and plasma progesterone concentration were measured 5 to 15 d after estrus. Data were analyzed by ANOVA with treatment, breed, and year and their 2-way interactions as independent variables. At breeding, HG heifers were heavier than LG heifers (419.9 vs.361.8 ± 7.5 kg; P < 0.01); ADG for the treatment period was 0.79 vs. 0.47 ± 0.04 kg/d (P < 0.01), respectively. In 2010 and 2011, 97.2% of heifers were cyclic by 21 d of breeding. Size of the ovary, preovulatory follicle, CL, and AFC did not differ between HG and LG, but preovulatory follicle diam. and ovarian length were greater (P ≤ 0.05) for MARC II vs. Angus heifers. Progesterone concentrations were less for LG vs. HG heifers (P ≤ 0.02), whereas CL volume was not affected by treatment or breed but was correlated positively with preovulatory follicle size (P < 0.01). Total AFC ranged from 5 to 49 and was correlated positively with ovarian volume but was not associated with fertility. A greater proportion of HG vs.LG heifers conceived within the first 21 d of the breeding period (64.4% vs. 49.2% ± 3.8%, respectively; P < 0.01), but overall pregnancy rate was not affected by treatment (83.0% vs. 77.7% ± 3.1%, respectively; P > 0.10). Pregnancy rate was 10% less (P < 0.01) for Angus vs. MARC II heifers. Developing beef heifers at a lesser ADG to a lighter BW (55% vs. 64% of mature BW) at breeding did not influence postweaning ovarian development or AFC or compromise pregnancy rate during the 47-d breeding period.

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Cattle genetically selected for twin ovulations and births (Twinner) exhibit increased ovarian follicular development, increased ovulation rate, and greater blood and follicular fluid IGF-1 concentrations compared with contemporary cattle not selected for twins (Control). Experimental objectives were to 1) assess relationships among aromatase (CYP19A1), IGF-1 (IGF1), IGF-2 receptor (IGF2R), and FSH receptor (FSHR) mRNA expression in small (≤5 mm) antral follicles and 2) determine their association with increased numbers of developing follicles in ovaries of Twinner females. Ovaries were collected from mature, cyclic (d 3 to 6) Twinner (n = 11), and Control (n = 12) cows at slaughter and pieces of cortical tissue were fixed and embedded in paraffin. Expression of mRNA was evaluated by in situ hybridization using (35)S-UTP-labeled antisense and sense probes for CYP19A1, FSHR, IGF1, and IGF2R mRNA. Silver grain density was quantified within the granulosa and theca cells of individual follicles (2 to 7 follicles/cow) by Bioquant image analysis. Follicles of Twinners tended to be smaller in diameter than Controls (1.9 ± 0.1 vs. 2.3 ± 0.1 mm; P = 0.08), but thickness of granulosa layer did not differ (P > 0.1) by genotype. Relative abundance of CYP19A1 (P < 0.01) and FSHR (P < 0.05) mRNA was greater in granulosa cells of Twinners vs. Controls, respectively, whereas IGF2R mRNA expression was less in both granulosa (P < 0.01) and theca (P < 0.05) cells in follicles of Twinners vs. Controls, respectively. Abundance of CYP19A1 mRNA in granulosa cells was correlated negatively with IGF2R mRNA expression in both granulosa (r = -0.33; P < 0.01) and theca (r = -0.21; P = 0.05) cells. Expression of IGF1 mRNA was primarily in granulosa cells, including cumulus cells, and its expression did not differ between Twinners vs. Controls (P > 0.10). Detected increases in CYP19A1 and FSHR, but not IGF1, mRNA expression along with decreases in IGF2R mRNA expression in individual follicles of Twinners support the hypothesis that increased follicular development and steroidogenesis in Twinner females result from increased extra-ovarian IGF-1 production. Furthermore, a reduction in follicular IGF2R mRNA expression accompanied by a reduction in receptor numbers would increase availability of free IGF-2 and its stimulation of follicular development in Twinners.