P. E. Lewis

Effect of peripheral concentrations of progesterone on follicular growth and fertility in ewes

The effects of progesterone (P4) on follicular growth and fertility in ewes were examined. In Experiment 1, 22 ewes received either one or three packets of P4 (5 g/packet) or an empty packet subcutaneously (sc) from Days 5 to 15 of the estrous cycle (estrus = Day 0). On Day 6, P4-treated ewes received 12.5 mg of prostaglandin F2 alpha. Follicles > or = 3 mm in diameter were observed via transrectal ultrasonography daily from Day 4 through estrus, corpora lutea (CL) were observed 5 to 7 d after estrus. Ewes with low (LOW; < or = 1 ng/ml; n = 5), intermediate (MED; >1 and <2 ng/ml; n = 10), or normal (NOR; > or =2 ng/ml; n = 7) P4 in jugular plasma on Days 7 through 15 differed in follicular development. The largest follicle at estrus was larger in ewes with LOW vs. MED and NOR P4 (7.8 +/- 0.3 vs. 6.9 +/- 0.2 mm; P < 0.05). Treatments differed in proportions of multiple-ovulating ewes, in which the oldest ovulatory follicle was first observed before Day 10 (LOW: 3 of 3, MED: 6 of 10, NOR: 0 of 5, respectively; P < 0.05). Estradiol was higher early in the treatment period in LOW ewes than in MED and NOR ewes (day x treatment; P < 0.05). In Experiment 2, ewes received 5 mg of P4 in corn oil (low progesterone [LP]; n = 51) or 2 ml of corn oil (CON; n = 49) sc every 12 hr on Days 6 through 14 of the estrous cycle before mating. LP ewes received 15 mg of prostaglandin F2 alpha on Day 6. Mean serum P4 on Days 7 through 15 was 0.6 +/- 0.1 ng/ml in LP and 1.9 +/- 0.1 ng/ml in CON ewes. Eleven LP and 12 CON ewes were scanned daily from Day 4 through mating, and in all ewes (n = 93), CL were counted 10 d after mating and embryos were counted at 25, 40, and 60 d of gestation. In multiple-ovulating ewes, day of cycle of appearance was earlier for the oldest (Day 6.1 +/- 0.8 vs. 10.4 +/- 0.8) but not second oldest (Day 11.7 +/- 1.0 vs. 12.2 +/- 0.9) ovulatory follicles in LP compared with CON ewes. The conception rate was lower in LP (72%) than in CON ewes (98%; P < 0.01). However, numbers of CL 10 d after mating, and in pregnant ewes, numbers of embryos 25 d after mating and lambs born, did not differ with treatment. In summary, low P4 increased the size of the largest follicles and the age of the oldest ovulatory follicles. Embryos resulting from the ovulation of older and younger follicles in the same ewe did not differ in their ability to survive.

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.

Factors contributing to the formation of experimentally-induced ovarian cysts in prepubertal gilts

Manipulation of an ovary during the follicular phase in cycling gilts or prepubertal gilts treated with PMSG and hCG results in formation of cysts on manipulated ovaries and corpora lutea (CL) of normal appearance on nonmanipulated ovaries. In contrast, cysts did not form after manipulation in luteal phase gilts. In the present experiment, daily administration of 50 mg progesterone to prepubertal gilts treated with PMSG and hCG established luteal phase concentrations of progesterone but did not lessen the incidence of manipulated-induced cysts. Number of cysts formed was associated with the number of follicles > or = 5 mm at manipulation, which was inversely related to serum concentrations of progesterone. Number of receptors for LH/hCG in follicular tissues did not differ between manipulated and nonmanipulated ovaries but was greater in granulosa (P < .05) and theca (P < .08) from follicles with diameters > or = 7 mm compared to 5 and 6 mm. Contents of estradiol, androstenedione, testosterone, progesterone and prostaglandins E2 and F2 alpha in follicular fluid, granulosa and theca were not different between follicles > or = 5 mm destined to form cysts. Profiles of progesterone and estradiol in peripheral serum and duration of luteal phase concentrations of progesterone were not different for gilts with induced cysts and gilts with CL. In conclusion, manipulation of follicles resulted in a failure to ovulate. Subsequent formation of cysts did not result from or result in a loss of steroidogenic function or the ability to bind LH to follicular receptors. These results demonstrate that the mechanism for ovulation is independent of other follicular processes, since ovulation can be disrupted without altering follicular steriodogenesis or subsequent luteinization.

Expression and Activation of Protein Kinase C Isozymes by Prostaglandin F2α in the Early- and Mid-Luteal Phase Bovine Corpus Luteum

Abstract Western blotting was used to identify the array of protein kinase C (PKC) isozymes expressed in the early (Day 4) and midcycle (Day 10) bovine corpus luteum (CL). PCKα, βI, βII, ϵ, and μ isozymes were detected in total protein samples prepared from both Day-4 and Day-10 corpora lutea. In contrast, specific antibodies for PKCγ, &eegr;, λ, and θ isozymes failed to detect protein bands in the luteal samples. PKCβII and ϵ isozymes were expressed differentially at these two developmental stages of the bovine CL. In the Day-4 luteal samples, PKCϵ was barely detectable; in contrast, in the Day-10 samples, the actin-corrected ratio for PKCϵ was 1.16 ± 0.13. This ratio was higher than the detected ratio for PKCβI and μ at this developmental phase of the CL (P < 0.01), but it was comparable with the ratio detected for the PCKα and βII. The amount of PKCβII was, although not as dramatic, also greater in the Day-10 CL (actin-corrected ratio was 0.85 ± 0.2) than in the Day-4 CL (0.35 ± 0.09 [P < 0.01]). The actin-corrected ratios for all other PKC isozymes, α (Day 4 = 0.93 ± 0.16, Day 10 = 0.97 ± 0.09), βI (Day 4 = 0.54 ± 0.073, Day 10 = 0.48 ± 0.74), and μ (Day 4 = 0.21 ± 0.042, Day 10 = 0.21 ± 0.38) were not different at these 2 days of the cycle. An experiment was designed to test whether activation of specific isozymes differed between CL that do or do not regress in response to PGF2α. Bovine CL from Day 4 and Day 10 of the estrous cycle were collected and 1 mm CL fragments were treated in vitro for 0, 2.5, 5, 10 or 20 min with PGF2α (0.1, 1.0, and 10 nM) or minimal essential medium-Hepes vehicle. Translocation of PKC from cytoplasm to membrane fraction was used as indication of PKC activation by PGF2α. Evidence for PKC activation was observed in both Day-4 and Day-10 luteal samples treated with 10 nM PGF2α. Therefore, if PKC, an intracellular mediator associated with the luteal PGF2α receptor, contributes to the lesser sensitivity of the Day-4 CL, it is likely due to the differential expression of the ϵ and βII isozymes of PKC at this stage and not due to an inability of the PGF2α receptor to activate the isozymes expressed in the early CL.