Bolzenius Jennifer K, Cushman Robert A, Perry George A
Department of Animal Science, South Dakota State University, Brookings, SD, USA.
USDA(1), ARS, U.S. Meat Animal Research Center, Clay Center, NE, USA.
Anim Reprod Sci. 2016 Aug;171:98-107. doi: 10.1016/j.anireprosci.2016.06.004. Epub 2016 Jun 9.
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.
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, and as uterine pH decreased, pregnancy success tended to increase.
在定时人工授精之前表现出发情的母牛,其精子向受精部位的运输增加,并且在授精后第6天胚胎质量得到改善。精子运输受子宫pH值影响,研究报告称发情开始时子宫pH值下降,但在排卵前必须恢复正常。因此,这些研究的目的是探讨发情开始前后子宫pH值调节的可能机制,并确定定时人工授精时的子宫pH值是否影响妊娠成功率。在实验1中,安格斯杂交肉牛(第1组和第2组分别为n = 40头和28头)采用PG 6天CIDR方案进行同期发情处理(第-9天注射PGF2α,第-6天注射GnRH并插入CIDR,第0天去除PGF2α和CIDR)。在去除CIDR时,根据卵泡大小对母牛进行分组,并在去除CIDR后的0、12、24、36、48、60小时(第1组)、72、84或96小时(第2组)采集子宫活检样本,从所有活检样本中提取总细胞RNA。通过HeatWatch发情检测系统监测发情情况。在实验2中,对两个牛群中的223头产后肉牛采用定时人工授精方案进行同期发情处理(牛群1:n = 97;CO - Synch加CIDR方案;牛群2:n = 126;Co - synch方案)。在人工授精时测定子宫pH值(牛群1和牛群2分别为n = 80头和63头),借助ESTROTECT发情检测贴片通过肉眼观察发情情况,并通过经直肠超声检查确定妊娠情况。在实验1中,在表现出发情的动物中,钠氢交换体亚型1、2和3的表达存在显著的(P<0.01)二次关系,在去除CIDR时表达最高,在发情开始时下降,然后在发情开始后再次上升。在未表现出发情的母牛中,不存在上述关系(P>0.46)。在实验2中,与未开始发情的母牛相比,在定时人工授精之前开始发情的母牛子宫pH值降低(P = 0.01)(分别为6.78±0.03和6.89±0.03),并且在观察到的pH值范围内,人工授精时的子宫pH值对妊娠成功率具有近似线性的影响。此外,与未开始发情的母牛相比,在人工授精之前开始发情的母牛妊娠成功率增加(P = 0.05)(52%对38%)。总之,在表现出发情的母牛中,去除CIDR后钠氢交换体亚型1、2和3的表达下降,但在未表现出发情的母牛中没有变化。此外,随着子宫pH值降低,妊娠成功率有增加的趋势(P = 0.076,逻辑回归)。因此,钠氢交换体亚型1、2和3似乎是发情开始前后子宫pH值的关键调节因子,并且子宫pH值的这种变化对妊娠成功至关重要。
在表现出发情的母牛中,去除CIDR后钠氢交换体亚型1、2和3的表达下降,但在未表现出发情的母牛中没有变化,并且随着子宫pH值降低,妊娠成功率有增加的趋势。