Ulbrich S E, Rehfeld S, Bauersachs S, Wolf E, Rottmayer R, Hiendleder S, Vermehren M, Sinowatz F, Meyer H H D, Einspanier R
Physiology-Weihenstephan, Technical University of Munich, Alles Hörsaalgebäude, 85350 Freising, Germany.
J Endocrinol. 2006 Feb;188(2):205-13. doi: 10.1677/joe.1.06526.
Nitric oxide synthases (NOS) account for the endogenous production of nitric oxide (NO), a small and permeable bioreactive molecule. NO is known to act as a paracrine mediator during various processes associated with female reproduction. In the present study, the mRNA expression of the endothelial (eNOS) and inducible (iNOS) NO synthases were examined in bovine oviduct epithelial cells (BOEC) during the oestrous cycle. In addition, eNOS and iNOS mRNA and protein were localised by in situ hybridisation and immunocytochemistry respectively. Furthermore, the effects of exogenously applied oestradiol-17beta and progesterone on NOS mRNA regulation were studied in a suspension culture of BOEC. The eNOS mRNA abundance was low around ovulation (day 0) and increased significantly until pro-oestrus (day 18) in the ampulla. Immunoreactive protein of eNOS was detected predominantly in endothelial cells as well as in secretory oviduct epithelial cells at pro-oestrus. The iNOS mRNA concentration was significantly reduced in the isthmus at pro-oestrus (day 18) and oestrus (day 0) compared with persistently high levels in the ampulla. By in situ hybridisation, specific iNOS transcripts were additionally demonstrated in the oviduct epithelium. Immunoreactive iNOS protein was localised in secretory epithelial cells as well as in the lamina muscularis. The in vitro stimulation showed that both NOS were stimulated by progesterone, but not by oestradiol-17beta. The region-specific modulated expression of eNOS and iNOS provides evidence for an involvement of endogenously produced NO in the regulation of oviductal functions.
一氧化氮合酶（NOS）负责内源性一氧化氮（NO）的生成，NO是一种小分子且具有渗透性的生物活性分子。已知NO在与雌性生殖相关的各种过程中作为旁分泌介质发挥作用。在本研究中，检测了发情周期中牛输卵管上皮细胞（BOEC）中内皮型（eNOS）和诱导型（iNOS）一氧化氮合酶的mRNA表达。此外，分别通过原位杂交和免疫细胞化学对eNOS和iNOS的mRNA及蛋白进行了定位。此外，在BOEC的悬浮培养中研究了外源性应用的17β-雌二醇和孕酮对NOS mRNA调节的影响。壶腹部eNOS mRNA丰度在排卵时（第0天）较低，直到发情前期（第18天）显著增加。在发情前期，eNOS的免疫反应性蛋白主要在内皮细胞以及分泌性输卵管上皮细胞中检测到。与壶腹部持续高水平相比，发情前期（第18天）和发情期（第0天）峡部的iNOS mRNA浓度显著降低。通过原位杂交，在输卵管上皮中还证实了特异性的iNOS转录本。免疫反应性iNOS蛋白定位于分泌性上皮细胞以及肌层。体外刺激表明，两种NOS均受孕酮刺激，但不受17β-雌二醇刺激。eNOS和iNOS的区域特异性调节表达为内源性产生的NO参与输卵管功能调节提供了证据。
