Caspase-3-mediated apoptosis and cell proliferation in the equine endometrium during the oestrous cycle.

Cell proliferation and apoptosis are hormone-dependent physiological processes involved in endometrial growth and regression. The aims of the present study were: (1) to evaluate endometrial cell proliferation using proliferating cell nuclear antigen (PCNA) expression; (2) to evaluate the induction of endometrial cell death by the expression of active caspase-3 and the apoptotic phenotype visualised by DNA fragmentation; and (3) to relate these observations to endometrial tissue dynamics in the equine endometrium throughout the oestrous cycle. Endometria were assigned to follicular and luteal phases based on ovarian structures and plasma progesterone. Cell proliferation and active caspase-3-mediated apoptosis were expressed in both phases of the oestrous cycle. In the luteal phase, PCNA expression was higher than in the follicular phase. Highest PCNA activity was noted in the luminal and glandular structures. Active caspase-3 staining was increased in luminal epithelium and deep glandular cells during the luteal phase. However, in the follicular phase, stromal cells showed greater active caspase-3 expression. Only a few apoptotic endometrial cells were detected by terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL) and these cells were mostly present in luminal and glandular structures. A simultaneous increase in DNA, cell proliferation and protein synthesis was observed in the endometrium during the mid-luteal phase. This suggests that cell hyperplasia occurs at the time the histotroph is needed for eventual embryo nourishment.

A case of blind vagina in a crossbred Lusitano mare.

This report describes a case of blind vagina diagnosed during a pre-breeding evaluation of a 5-year-old crossbred Lusitano mare. This mare was mated twice during the previous breeding season but remained open. Clinical evaluation revealed the existence of follicles in the ovaries and an enlarged uterus. An ultrasound examination showed that a granular free-floating fluid distended the uterus. No connection was detected between the uterus and the vagina and a presumptive diagnosis of congenital vaginal obstruction was raised. On vaginal examination it was noticed that the vagina was short in depth and ended as a blind pouch. The typical cervical morphology was not observed. Careful evaluation of the clinical situation, both during the follicular and the luteal phases of the mare's oestrous cycle, lead to a diagnosis of segmental aplasia of the cranial vagina. The mare had a normal karyotype on cytogenetic examination [64XX].

Progesterone receptors and proliferating cell nuclear antigen expression in equine luteal tissue.

Steroid hormones act via specific receptors, and these play an important physiological role in the ovary. The objective of this study was to evaluate the cellular distribution of progesterone receptors and their staining intensity in different equine luteal structures during the breeding season, as well as their relationship to luteal cell composition, cell proliferation pattern and plasma progesterone (P4) concentration. There was an increase in proliferating cell nuclear antigen (PCNA) expression in large luteal cells from the corpus hemorrhagicum (CH) to mid-luteal phase, followed by a decrease toward the late luteal stage. In the CH, the number of large luteal cells was lower than in other structures. Only large luteal cells showed positive staining for P(4) receptors. An increase in staining intensity for P(4) receptors was observed between CH and mid-phase corpus luteum, and CH and late-phase corpus luteum. Synthesis of P(4) started at a very early stage of the luteal structure and was accompanied by an increase in P(4) receptors and PCNA expression, and proliferation of large luteal cells, until mid-luteal phase. These data suggest that large luteal cells might play an important role in the regulation or synthesis of P(4) in equine luteal structures.

Influence of puerperal uterine infection on uterine involution and postpartum ovarian activity in dairy cows.

This article presents the results of a clinical trial designed to study the effect of puerperal uterine infection on uterine involution and on ovarian activity in dairy cows, monitored twice weekly from parturition until the sixth week postpartum (wpp). Infection significantly retarded uterine involution assessed by the uterine body diameter and a score of intrauterine fluid volume (IUFV). By the sixth wpp, cows with normal puerperium (controls) and cows that showed mild puerperal endometritis had similar uterine body diameter and IUFV, indicating spontaneous recovery within the postpartum voluntary waiting period. However, in cows with severe puerperal endometritis, although uterine body diameter had regressed to pregravid size, IUFV remained significantly higher than in control and mild endometritis cows, indicating that chronic endometritis was established. The IUFV score was positively and significantly correlated with uterine swab bacterial growth density and allowed diagnosis of endometritis after the third wpp. Cows with mild or severe endometritis had a significantly higher prevalence and persistence of pathogenic bacteria (Escherichia coli, Actinomyces pyogenes, Gram-negative anaerobes - GNA) than controls. Actinomyces pyogenes was associated to GNA in 74% of isolations. Ovarian activity measured by ultrasound scanning of the ovaries and plasma progesterone (P4) concentrations was more abnormal (prolonged anoestrus, prolonged luteal phases and ovarian cysts) in cows with severe endometritis than in controls.

Treatment of unobserved oestrus in a dairy cattle herd with low oestrous detection rate up to 60 days post-partum.

The efficiency of treatments for unobserved oestrus and their effect on the reproductive performance of a dairy cattle herd with low oestrous detection rate till 60 days post-partum (dpp), attributed to the declivous and slippery concrete floor were investigated. The herdsman requested advice in order to improve the mean days open of the herd, but no investments were allowed because a new unit was about to be built. Due to the low oestrus detection rate of the herd, the breeding policy was to inseminate at the first detected post-partum oestrus. Cows were examined at 20-30 dpp to assess uterine involution, ovarian activity and prevalence of reproductive disorders and, at 60 dpp if no previous oestrus was detected. Each examination included palpation per rectum, ultrasound scanning and collection of a blood sample for plasma progesterone (P4) measurement. Cows with unobserved oestrus till 60 dpp were allocated either to a treatment group (n=139) or to a control group (n=139). Three treatments were used: (a) injection of PGF(2 alpha) (PG) upon detection of a corpus luteum (CL; n = 30), cows not observed in oestrus being re-injected 11-12 days later. AI was at oestrus; (b) PRID (n=35) or Crestar (n=74) devices kept in situ for 12 and 9 days, respectively, were associated to an injection of PG and of equine chorionic gonadotrophin (eCG) at device removal. Cows were double-fixed time-inseminated at 48 and 72 h after device removal. All treated cows were examined at 48-72 h after treatment to confirm oestrus. The percentage of cows detected in oestrus up to 60 dpp remained unchanged through the trial (35 and 47% for years before intervention: 1994-95; 51 and 48% for years of intervention: 1996-97). In contrast, the oestrous detection rate was high both in treated (93%) and control (100%) cows. This possibly resulted from an improvement in the oestrous detection efficiency of the herd's personnel and from examination of cows at 48-72 h after treatment. Treated and control cows had identical conception rate (CR; 36 and 37%, respectively) and reproductive performance. However, the mean days open of the herd in 1996 was significantly improved in comparison with previous years (mean +/- SEM: 134 +/- 6, 126 +/- 5, 110 +/- 4 and 114 +/- 5 days, for years 1994, 1995, 1996 and 1997, respectively, p < 0.05, ANOVA). Conception rate to AI up to 40 dpp was significantly reduced, compared with the period between 60 and 100 dpp but, mean days open were significantly improved in cows inseminated up to 60 dpp, compared with thereafter (p < 0.05).