Although the decrease of progesterone in serum and in luteal tissue during luteal regression is well characterized, relatively little is known about changes in proteins produced by the corpus luteum during this time. The first objective was to examine changes in patterns of protein secretion that might be associated with functional and structural luteal regression. The second objective was to characterize the expression of two major secretory products of regressing corpora lutea. Thirty normally cyclic heifers were randomly assigned at day 15–16 of the oestrous cycle (oestrus = day 0) to be ovariectomized at 0 h (no PGF_2α; n = 5) or at 4, 8, 12, 24 or 48 h after PGF_2α-induced luteal regression (n = 5 per time point). Total cellular RNA was isolated from tissue frozen at the time of ovariectomy. Thin slices (< 1 mm) of tissue were placed in methionine-deficient minimum essential media with [³⁵S]methionine and placed in a humidified CO₂ incubator at 38°C. Media and tissues were collected 6 h later. Changes in profiles of secreted proteins were analysed by one-dimensional SDS-PAGE. A number of proteins (relative molecular mass ranging from 14 300 to 200 000) were produced by luteal tissue at each time point (0–48 h). The major secretory proteins had relative molecular masses of approximately 21 500, 28 200, 43 700 and 46 000. Secretion of the relative molecular mass 46 000 protein(s) increased (P < 0.05) between 4 and 24 h after PGF_2α injection compared with the 0 h group. Western blot analyses with either tissue inhibitor of metalloproteinase-1 or tissue inhibitor of metalloproteinase-2 antisera detected immunoreactive proteins of relative molecular mass 28 200 and 21 500, respectively. Concentrations of mRNA encoding tissue inhibitor of metalloproteinases-1 increased (P < 0.01) by 8 h after PGF_2α injection, remained stable (P > 0.20) through 24 h and decreased (P < 0.05) by 48 h after PGF_2α. Concentrations of tissue inhibitor of metalloproteinases-2 mRNA were highest (P < 0.05) at 8 h after PGF_2α injection and lowest (P < 0.05) 48 h following induction of luteolysis. In summary, the profile of luteal protein production changed during luteolysis and two secretory products (tissue inhibitor of metalloproteinases-1 and -2) were identified. Metalloproteinase inhibitors may have an important role in tissue remodelling during structural luteolysis.

A study was conducted to examine the effects of gonadotropins on ovarian follicular development and differentiation in GnRH agonist (GnRHa)-treated cattle. Holstein cows were allotted into two pre-treatment groups: controls (n = 5) and GnRHa-treated (n = 9). Ovaries were removed from control cows on day 5 following a synchronized estrus. Treatment with GnRHa resulted in follicular arrest at <5 mm. Following follicular arrest, GnRHa-treated cows received a constant infusion of FSH for 96 h (GnRHa/FSH), with a randomly selected subset receiving hourly pulses of LH in addition to FSH during the last 48 h of infusion (GnRHa/FSH + LH). At the end of infusion, ovaries were removed, follicles were counted and measured, and follicular fluid samples were collected from large follicles (>10 mm). Differences in expression of mRNA for LH receptor, FSH receptor, cytochrome P450 side-chain cleavage, 3β-hydroxysteroid dehydrogenase, cytochrome P450 17α-hydroxylase (P450c17) and cytochrome P450 aromatase were determined in large follicles using in situ hybridization. The number of large follicles did not differ between GnRHa/FSH-treated and GnRHa/FSH + LH-treated cows (P = 0.64), but was greater than control animals (P ≤ 0.004). Follicular fluid concentrations of estradiol-17β and androstenedione were highest in GnRHa/FSH + LH-treated cows (P ≤ 0.04), intermediate in control cows, and lowest in GnRHa/FSH-treated cows. Hybridization intensity of P450c17 was greater in GnRHa/FSH + LH-treated versus control or GnRHa/FSH-treated cows (P ≤ 0.03). These results indicate that while FSH can support bovine follicular growth >10 mm, LH increases androgen production and expression of P450c17.

Summary. Non-lactating, multiparous dairy cows diagnosed as having cysts by palpation per rectum were used. Cysts were induced with oestradiol-17β (15 mg) and progesterone (37·5 mg) dissolved in ethanol and injected s.c. twice daily for 7 days. Following initial diagnosis of cysts, ovaries were exposed by midventral laparotomy, and the perimeter of the base of each cyst was marked with subepithelial injections of charcoal. Ovaries were removed from cows by transvaginal incision at 10 days (Group 1; N = 8), 20 days (Group 2; N = 8), or 40 days (Group 3; N = 7) after marking of cysts. Ovaries were examined for structures present and their relationship to the marked site. Corpora lutea with ovulation papilla were present in 7/23 cows (1/8, 4/8 and 2/7, for Groups 1, 2 and 3, respectively). In these 7 cows, corpora lutea were at a site different from the original structure that was marked. Marked structures persisted for the duration of the experimental period in 1 and 2 cows, in Groups 1 and 3, respectively. In the remaining 13 cows, new large follicular structures (cysts) were present at a site other than that marked with charcoal. These structures developed on the ovary contralateral to the one originally marked in 9 of 13 cows. Cysts are therefore dynamic in nature and may persist or may be replaced by others.

Keywords: ovary; follicular cysts; turnover rate; dairy cattle

Summary. Two experiments were conducted to (1) investigate developmental endocrinology of ovarian follicular cysts (cysts) in cattle and (2) evaluate effects of cysts on hypothalamic and hypophysial characteristics. Cysts were induced with oestradiol-17β (15 mg) and progesterone (37·5 mg) dissolved in alcohol and injected s.c. twice daily for 7 days. Cysts were defined as the presence of follicular structures (which may or may not have been the same structure) of 2·0 cm in diameter or greater that were present for 10 days without ovulation and corpus luteum development.

In Exp. 1, 22 non-lactating, non-pregnant Holstein cows were allocated to 3 groups. Beginning on Day 5 (oestrus = Day 0) of the oestrous cycle, 7 cows (Controls) were treated with twice daily s.c. injections of ethanol (2 ml/injection) for 7 days. Luteolysis was then induced with PGF-2α and blood samples were collected daily every 15 min for 6 h from the morning after the PGF-2α injection (Day 13) until oestrus. Steroids to induce cysts were injected as previously described into the remaining cows (N = 15). Three blood samples were collected at 15-min intervals every 12 h throughout the experimental period. Additional blood samples were collected every 15 min for 6 h on a twice weekly basis. After steroid injections, follicular and luteal structures on ovaries were not detected via rectal palpation for a period of 36 ± 4 days (static phase). Then follicles developed which ovulated within 3–7 days (non-cystic; N = 7) or increased in size with follicular structures present for 10 days (cystic; N = 8). Mean (± s.e.m.) concentrations of LH, FSH, oestradiol-17β and progesterone in serum remained low and were not different during the static phase between cows that subsequently developed cysts or ovulated. During the follicular phase, mean serum concentration of LH (ng/ml) was higher (P < 0·1) in cows with cysts (2·9 ± 0·2) than in cows without cysts (1·1 ± 0·1) or control cows (1·4 ± 0·2). In addition, LH pulse frequency (pulses/6 h) and amplitude (ng/ml) were higher (P < 0·1) in cows with cysts (3·6 ± 0·3 and 2·2 ± 0·3, respectively) than in non-cystic (2·3 ± 0·2 and 1·0 ± 0·2, respectively) and control (1·8 ± 0·1 and 1·1 ± 0·2, respectively) groups during the follicular phase.

In Exp. 2, 20 non-lactating, non-pregnant dairy cows were used: 15 cows received exogenous steroids as previously described. Hypothalamic and hypophysial tissues were collected after diagnosis of cystic structures in 11 cows (cystic group). The remaining 4 cows in the steroid-treated group ovulated and were assigned to the control group in addition to 5 non-steroid treated cows. Hypothalamic and hypophysial tissues were collected during the late-luteal phase (Days 16–18) from these control cows (N = 9). Anterior pituitary concentrations (μg/g) of LH (60·5 ± 11·0, 44·6 ± 11·7), FSH (30·2 ± 4·0, 22·1 ± 4·6) and receptors for GnRH (17·2 ± 2·2, 23·4 ± 2·6 m × 10⁻¹⁰/mg protein) did not differ between cows with cysts and control cows, respectively. Content of GnRH (ng) in the combined preoptic area and hypothalamus proper was higher (P < 0·05) in control cows (37·7 ± 6·6) than cows with cysts (18·6 ± 6·1). In the pituitary stalk median eminence, GnRH content (ng) tended to be higher (P ≥ 0·1) in cows with cysts (38·5 ± 9·6) compared with control (21·1 ± 15·2) cows.

Secretory patterns (mean concentration, pulse frequency and amplitude) of LH were therefore increased during the follicular phase in cows which developed cysts compared to cows which subsequently ovulated. In addition, hypothalamic GnRH content, but not pituitary characteristics, appeared to be altered in cows with cysts.

Keywords: ovary; follicular cysts; dairy cattle; gonadotrophins; hypothalamus; pituitary

Luteal lifespan is short after first postpartum ovulation in early-weaned beef cows unless cows are pretreated with a progestogen. Regression of the short-lived corpus luteum in the postpartum beef cow is due to a premature release of prostaglandin F_2α (PGF_2α) from the uterus. The premature release of PGF_2α may be mediated through lower concentrations of receptors for progesterone, higher concentrations of oxytocin receptors, or both, in the endometrium. Thirty-one beef cows were randomly assigned to four groups at parturition. Calves from cows assigned to the short cycle group (n = 6; control) and the short cycle/endometrium group (n = 10) were weaned at 30–32 days post partum. Cows in the normal cycle group (n = 5; control) and the normal cycle/endometrium group (n = 10) received norgestomet implants for 9 days beginning 21–23 days post partum and calves were weaned at implant insertion. Duration of oestrous cycle (x ± sem; P < 0.01) following first postpartum ovulation for the short cycle group was 11.5 ± 1.9 days compared with 18.8 ± 0.6 days for the normal cycle group. On day 5 following first postpartum ovulation, cows in the short cycle/endometrium and the normal cycle/endometrium groups were hysterectomized and endometrial tissue collected for measurement of progesterone and oxytocin receptors. Mean number of total progesterone receptors per cell was lower (P < 0.05) in the short cycle/endometrium group than in the normal cycle/endometrium group. Mean concentration of oxytocin receptors (fmol mg⁻¹ protein) in the short cycle/endometrium group was higher (P < 0.05) than that in the normal cycle/endometrium group. In conclusion, uterine receptor populations for progesterone and oxytocin may influence the timing of PGF_2α secretion during short oestrous cycles.