Ultrasound-guided aspiration of dominant follicles (≥25 mm) followed by luteinization and progesterone production during the estrous cycle in mares.

The present study was designed to evaluate luteinization rates subsequent to aspiration of dominant follicles (≥25 mm) in the absence of a functional CL (progesterone <1 ng/mL) and characterize the temporal changes in plasma concentrations of progesterone following aspiration-induced luteinization during the estrous cycle in mares. A total of 29 estrous cycles involving 15 mares in a cross-over design were randomly assigned to five groups: 1) ASP-F≥25 mm (n = 6; follicle aspiration 25-29 mm), 2) ASP-F≥30 mm (n = 6; follicle aspiration 30-34 mm), 3) ASP-F≥35 mm (n = 6; follicle aspiration 35-40 mm), 4) ASP-F≥40 (n = 6; follicle aspiration ≥40), and 5) Control (n = 5; spontaneous ovulation or no follicle aspiration). Subsequent to ovulation (Day 0), PGF was administered to all groups on Day 5, blood samples were collected daily and aspiration of the dominant follicle was done using ultrasound-guided transvaginal follicle needle puncture. Among the follicle aspirations groups 25-29, 30-34, 35-39, and ≥40 mm, the luteinization rates were not different (P > 0.05) at 83, 67, 83, and 100%, respectively. Correspondingly, progesterone concentrations increased (>2 ng/mL) by approximately 6, 7, 5, and 4 d after aspiration, respectively, which were delayed (P < 0.05) in the 25-29 and 30-34 mm follicle aspiration groups compared to 2 d after ovulation in the control group. Thereafter, progesterone reached maximal concentrations (10-11 ng/mL) as averaged over all aspiration groups but were lower (P < 0.05) compared to the mean maximal concentration (18 ng/mL) in the control group. Subsequently, there was a decrease in progesterone concentrations (<2 ng/mL) in response to luteolysis, which was delayed (P < 0.05) in the aspiration groups over Days 16-20 compared to Day 15 in the control group. Despite this discrepancy, the mean length of the interovulatory intervals were not different (P > 0.05) among groups on Day 23. Thus, the present study provided novel information that the luteinization rate is relatively high (83%) and consistent following aspiration of dominant follicles (≥25 mm) in the absence of a functional CL and that the increase in progesterone reaches sustainable progestational concentrations (≥2 ng/mL) in accord with the length of the estrous cycle that may potentially support development and maintenance of early pregnancy in recipient mares involved in an embryo transfer program.

Administration of 2.5 mg of estradiol followed by 1,500 mg of progesterone to anovulatory mares promote similar uterine morphology, hormone concentrations and molecular dynamics to those observed in cyclic mares.

To test the hypothesis that the administration of 2.5 mg of estradiol benzoate (EB) followed by 1500 mg of long acting progesterone (LA P4) causes similar uterine changes and molecular dynamics in anovulatory mares to those observed in cyclic ones, we evaluated the changes of estrogen (ERα and ERß) and progesterone receptors (PR) in anestrous, transitional and cyclic mares by RT-qPCR and immunohistochemistry. In addition, we evaluated uterine edema, tonus and estrogens and progesterone plasma profile. Endometrial biopsies were taken from anestrous and transitional mares immediately before EB injection, 48 h after EB administration and five days after LA P4 was given. In cyclic mares, biopsies were collected at estrus and at five days after ovulation. Similar estrogen peaks were achieved after the injection of the single EB dose between treated and cyclic groups, as well as maximum uterine edema. Uterine tone was increased to diestrus levels after administration of 1500 mg of LA P4. Changes in relative abundance of transcripts for PR, ERα and ERß when progesterone stimulated endometrium was compared to estrogen stimulated endometrium were similar between cyclic and non-cyclic treated mares. However, apparent decreased PR in the endometrial glandular epithelium was not observed in non-cyclic mares five days after LA P4 administration as observed at five days after ovulation in cyclic mares. The protocol produced similar endometrial edema, uterine tonus and changes in relative abundance of PR, ERα and ERß transcripts to those observed in cyclic mares during late estrus and early diestrus, as well as similar estradiol and estrogen conjugate plasma concentrations.

Effect of age and endometrial degenerative changes on uterine blood flow during early gestation in mares.

The present experiment was divided into three studies to investigate the effect of age and endometrial degeneration on uterine blood flow of mares. In study 1, the influence of semen infusion and conception was evaluated using noninseminated (non-AI), inseminated nonpregnant (AI-NP), and inseminated pregnant (AI-P) mares (n = 7 mares/group). In study 2, the effect of age was investigated using young (≤6 years), adult (from 8 to 12 years), and old (≥15 years) pregnant mares (n = 7 mares/group). In study 3, uterine blood flow was also characterized in pregnant mares (n = 7 mares/group) with minimal, moderate, or severe endometrial degenerative changes (GI, GII, and GIII, respectively). Uterine vascular perfusion and pulsatility index from mesometrium attachment arteries were recorded daily from Day 0 (day of ovulation) to Day 12 and between Days 0 and 20 in nonpregnant and pregnant mares. Analysis according to the position of the embryo (uterine horn with embryo vs. opposite horn) was performed from Day 12 until Day 20. In study 1, increased uterine vascular perfusion and decreased pulsatility index were detected (P < 0.001) between Days 3 and 5 in non-AI and AI-P mares, whereas the uterine vascular perfusion of AI-NP mares did not change (P > 0.05) throughout the experiment. In study 2, the vascular perfusion of the uterine horn with embryo was higher (P < 0.001) than in the opposite uterine horn from Day 12 until Day 20 in both young and adult mares. With exception of Day 15, both uterine horns of old mares showed similar (P > 0.1) vascular perfusion from Day 12. In study 3, vascular perfusion of both uterine horns was lower (P < 0.001) in GIII mares than in GI and GII mares. After Day 15, the scores for uterine vascular perfusion were consistently greater (P < 0.001) in the uterine horn with embryo than in the opposite horn in GI mares. Results showed, for the first time, the reduced blood flow of the gravid uterus during early gestation in older mares and in mares with significant endometrial degeneration.