Laboratory of Animal Breeding and Reproduction, Department of Animal Science, Research Faculty of Agriculture, Hokkaido University, Hokkaido 060-8589, Japan.
Dairy Cattle Group, Konsen Agricultural Experiment Station, Hokkaido Research Organization, Nakashibetsu, Hokkaido 086-1135, Japan.
J Dairy Sci. 2018 Sep;101(9):8396-8400. doi: 10.3168/jds.2017-14251. Epub 2018 Jun 21.
In ruminants, IFN-tau (IFNT) is a pregnancy recognition signal secreted by the embryonic trophectoderm before implantation, and it induces the expression of IFN-stimulated genes (ISG) in the uterine endometrium and blood leukocytes. The expression of ISG in blood leukocytes could indicate the presence of a viable conceptus before return of the next estrus; however, expression levels have high variation for confirming pregnancy. We hypothesized that the secreted IFNT in the uterus would affect ISG expression in cervical and vaginal tissues because they are directly adjacent to the uterus. To prove the hypothesis, we investigated the expression of 3 ISG (ISG15, MX1, and MX2) in cervical and vaginal mucosal membranes collected from pregnant (n = 12) and nonpregnant (n = 11) lactating Holstein cows at 17 to 18 d after artificial insemination. Mucosal membrane samples of the cervical canal near the external os (cervix) and deep vaginal wall surrounding the external os (vagina) were collected separately by simply scraping with a curette on d 17 or 18 of pregnancy (d 1 = ovulation), at which time IFNT secretion into the maternal uterus is maximal. After pregnancy diagnosis on d 30 and 60, separately collected samples confirmed as pregnant and nonpregnant were used for evaluation of the expression of IFN-stimulated protein 15 kDa (ISG15) and myxovirus-resistance protein 1 and 2 (MX1, MX2) with quantitative real-time PCR. The collected mucosal membrane samples from cervix contained mostly cell clots showing membrane structure and a low content of blood cells. The expression levels of all 3 genes were significantly increased in pregnant cows compared with nonpregnant cows in both cervical and vaginal samples. These results suggest that increased expression of ISG in the cervix and vagina is a pregnancy-associated phenomenon and is highly affected by IFNT secreted from the conceptus through the uterus.
在反刍动物中,IFN-τ(IFNT)是胚胎植入前滋养外胚层分泌的妊娠识别信号,它诱导子宫子宫内膜和血液白细胞中 IFN 刺激基因(ISG)的表达。血液白细胞中 ISG 的表达可以表明在下次发情前存在有活力的胚胎;然而,表达水平存在很大的差异,难以确认妊娠。我们假设子宫中分泌的 IFNT 会影响宫颈和阴道组织中的 ISG 表达,因为它们与子宫直接相邻。为了证明这一假设,我们在人工授精后 17 至 18 天,调查了怀孕(n=12)和未怀孕(n=11)泌乳荷斯坦奶牛的宫颈和阴道黏膜中 3 种 ISG(ISG15、MX1 和 MX2)的表达。在怀孕第 17 天或第 18 天(排卵第 1 天),通过简单地用刮匙刮取靠近外口(宫颈)的宫颈管和环绕外口的阴道深壁的黏膜样本,分别收集宫颈和阴道黏膜样本。此时 IFNT 分泌到母体子宫内达到最大值。在妊娠诊断后的第 30 天和第 60 天,分别收集的被证实怀孕和未怀孕的样本用于评估干扰素刺激蛋白 15kDa(ISG15)和粘液病毒抗性蛋白 1 和 2(MX1、MX2)的表达,使用定量实时 PCR。从宫颈采集的黏膜膜样本主要含有细胞凝块,表现出膜结构,并且含有少量血细胞。与未怀孕的奶牛相比,怀孕的奶牛在宫颈和阴道样本中,所有 3 种基因的表达水平均显著升高。这些结果表明,宫颈和阴道中 ISG 的表达增加是一种妊娠相关现象,并且高度受到来自胚胎的通过子宫分泌的 IFNT 的影响。