Institute of Veterinary Anatomy, University of Zurich, Winterthurerstrasse 260, Zurich, 8057, Switzerland.
Reprod Biol Endocrinol. 2012 Aug 29;10:66. doi: 10.1186/1477-7827-10-66.
Transient receptor potential channel type 6 (TRPV6) and Calbindin-D9k (CaBP-9k) are involved in the active calcium (Ca2+) transport mechanism in many tissues including placenta and uterus, suggesting a role in the establishment and maintenance of pregnancy. Moreover, TRPV6 and CaBP-9k seem to support the materno-fetal Ca2+ transport that is crucial for fetal Ca2+ homeostasis, bone growth and development. However, it is unknown if these proteins are also involved in the aetiology of pathologies associated with parturition in cows, such as retained fetal membranes (RFM). The aim of the current study was to create an expression profile of uterine and placentomal TRPV6 and CaBP-9k mRNAs and proteins during pregnancy and postpartum in cows with and without fetal membrane release.
Uteri and placentomes of 27 cows in different stages of pregnancy and placentomes of cows with and without RFM were collected. Protein and mRNA expression of TRPV6 and CaBP-9k was investigated by real-time PCR, immunohistochemistry and Western blot.
In the uterine endometrium, highest TRPV6 and CaBP-9k expression was found in the last trimester of pregnancy, with a particular increase of protein in the glandular epithelium. In the placentomes, a gradual increase in TRPV6 mRNA was detectable towards parturition, while protein expression did not change significantly. Placentomal CaBP-9k expression did not change significantly throughout pregnancy but immunohistochemistry revealed an increase in staining intensity in the maternal crypt epithelium. Immunohistochemical, stronger placental CaBP-9k signals were seen in animals with RFM compared to animals with an undisturbed fetal membrane release, while protein levels, measured by Western blot analyses did not change significantly.
The results of the present study demonstrate a dynamic expression of TRPV6 and CaBP-9k during pregnancy in the bovine uterine endometrium and placentomes, suggesting a functional role for these proteins in Ca2+ metabolism during pregnancy. The temporal and spatial expression patterns indicate that TRPV6 and CaBP-9k may be involved in materno-fetal Ca2+ transport, mainly through an interplacentomal transport, and that both proteins may participate in physiological processes that are crucial for fetal and placental development. However, neither TRPV6 nor CaBP-9k seem to be causative in the retention of fetal membranes.
瞬时受体电位通道 6(TRPV6)和钙结合蛋白-D9k(CaBP-9k)参与包括胎盘和子宫在内的许多组织中的主动钙(Ca2+)转运机制,这表明它们在妊娠的建立和维持中起作用。此外,TRPV6 和 CaBP-9k 似乎支持母体-胎儿 Ca2+转运,这对于胎儿 Ca2+稳态、骨骼生长和发育至关重要。然而,目前尚不清楚这些蛋白是否也参与与奶牛分娩相关的病理学的发病机制,例如胎衣滞留(RFM)。本研究的目的是在有和没有胎膜释放的奶牛中,建立妊娠和产后子宫和胎盘中 TRPV6 和 CaBP-9k mRNA 和蛋白的表达谱。
收集了 27 头处于不同妊娠阶段的奶牛的子宫和胎盘中,以及有和没有 RFM 的奶牛的胎盘中的 TRPV6 和 CaBP-9k 的蛋白和 mRNA 表达情况,通过实时 PCR、免疫组织化学和 Western blot 进行了研究。
在子宫子宫内膜中,TRPV6 和 CaBP-9k 的表达在妊娠末期最高,特别是在腺上皮中蛋白表达增加。在胎盘中,TRPV6 mRNA 逐渐增加,直到分娩,但蛋白表达没有显著变化。胎盘中 CaBP-9k 的表达在整个妊娠过程中没有显著变化,但免疫组织化学显示母性隐窝上皮的染色强度增加。与胎膜正常释放的动物相比,有 RFM 的动物的胎盘 CaBP-9k 的免疫组织化学信号更强,但通过 Western blot 分析测量的蛋白水平没有显著变化。
本研究的结果表明,在牛的子宫子宫内膜和胎盘中,TRPV6 和 CaBP-9k 在妊娠期间表现出动态表达,这表明这些蛋白在妊娠期间的 Ca2+代谢中具有功能作用。时空表达模式表明,TRPV6 和 CaBP-9k 可能参与母体-胎儿 Ca2+转运,主要通过胎盘间转运,并且这两种蛋白都可能参与对胎儿和胎盘发育至关重要的生理过程。然而,TRPV6 和 CaBP-9k 似乎都不是胎衣滞留的原因。