Expression of Na+/H+ exchanger isoforms 1, 2, 3, and 4 in bovine endometrium and the influence of uterine pH at time of fixed-time AI of pregnancy success

Abstract

Cows that exhibit estrus prior to fixed-time AI had increased sperm transport to the site of fertilization, and improved embryo quality on d 6 after insemination. Sperm transport is influenced by uterine pH, and research has reported that uterine pH decreased at onset of estrus, but must return to normal prior to ovulation. Therefore, the objectives of these studies were to investigate a possible mechanism for the regulation of uterine pH around the onset of estrus, and to determine if uterine pH at time of fixed-time AI influenced pregnancy success. In experiment 1, Angus-cross beef cows (n=40 and 28 in rep. 1 and 2, respectively) were synchronized with the PG 6-day CIDR protocol (PGF2α on d -9, GnRH and insertion of a CIDR on d -6, and PGF2α and CIDR removal on d 0). Cows were blocked by follicle size at time of CIDR removal, and uterine biopsies were collected at 0, 12, 24, 36, 48, 60 (Rep. 1), 72, 84, or 96h (Rep2) after CIDR removal, and total cellular RNA was extracted from all biopsies. Estrus was monitored by the HeatWatch Estrous Detection System. In experiment 2, 223 postpartum beef cows in 2 herds were synchronized with a fixed-time AI protocol (herd 1: n=97; CO-Synch plus CIDR protocol; herd 2: n=126; Co-synch protocol). Uterine pH was determined at time of AI (n=80 and 63 for herd 1 and 2, respectively), and estrus was monitored by visual estrus detection with the aid of an ESTROTECT estrous detection patches, and pregnancy was determined by transrectal ultrasonography. In experiment 1, there was a significant (P<0.01), quadratic relationship in expression of Na+/H+ exchanger isoforms 1, 2, and 3 among animals that exhibited estrus, with expression greatest at time of CIDR removal, decreasing to the onset of estrus, and then increasing again following the onset of estrus. Among cows that did not exhibit estrus, the preceding relationship did not exist (P>0.46). In experiment 2, cows that had initiated estrus prior to fixed-time AI had decreased (P=0.01) uterine pH compared to cows that did not initiate estrus (6.78±0.03 and 6.89±0.03, respectively), and uterine pH at AI had an approximately linear effect on pregnancy success within the observed pH range. Furthermore, cows that initiated estrus prior to AI had increased (P=0.05) pregnancy success (52% vs. 38%) compared to cows that had not initiated estrus. In summary, expression of Na+/H+ exchanger isoforms 1, 2, and 3 decreased after CIDR removal among cows that exhibited estrus, but did not change among cows that did not exhibit estrus. Additionally, as uterine pH decreased pregnancy success tended to increase (P=0.076, logistics regression). Thus, Na+/H+ exchanger isoforms 1, 2, and 3 appear to be key regulators of uterine pH around the onset of estrus, and this change in uterine pH is critical for pregnancy success. Summary sentenceExpression of Na+/H+ exchanger isoforms 1, 2, and 3 decreased after CIDR removal among cows that exhibited estrus, but did not change among cows that did not exhibit estrus, and as uterine pH decreased, pregnancy success tended to increase.