Kautz Ewa, de Carvalho Papa Paula, Reichler Iris M, Gram Aykut, Boos Alois, Kowalewski Mariusz P
Institute of Veterinary Anatomy, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 260, CH-8057, Zurich, Switzerland.
Department of Surgery, Faculty of Veterinary Medicine and Animal Sciences, Sector of Anatomy, University of Sao Paulo, Sao Paulo, Brazil.
Reprod Biol Endocrinol. 2015 Aug 5;13:85. doi: 10.1186/s12958-015-0066-4.
The uterine response to the presence of embryos is poorly understood in the domestic dog (Canis familiaris). The intimate embryo-maternal cross-talk, which begins following the hatching of blastocysts and embryo attachment leads to strong structural and functional remodelling of the uterus. A part of this process is decidualisation, comprising morphological and biochemical changes that result in formation of maternal stroma-derived decidual cells. These are an integral part of the canine placenta materna, which together with the maternal vascular endothelium are the only cells of the canine endotheliochorial placenta able to resist trophoblast invasion. These cells are also the only ones within the canine placenta expressing the progesterone receptor (PGR). Understanding the decidualisation process thus appears essential for understanding canine reproductive physiology.
Here, we investigated the capability of canine uterine stromal cells to decidualise in vitro, thereby serving as a canine model of decidualisation. A dbcAMP-mediated approach was chosen during a time course of 24 - 72 h. Tissue material from six (n = 6) healthy, dioestric bitches was used (approximately 2 weeks after ovulation). Cells were characterized by differential staining, nearly 100 % of which were vimentin-positive. Scanning and transmission electron microscope analyses were applied, and morphological changes were recorded with a live cell imaging microscope. Expression of several decidualisation markers was investigated.
The in vitro cultured stromal cells acquired characteristics of decidual cells when incubated with 0.5 mM dbcAMP for 72 h. Their shape changed from elongated to rounded, while ultrastructural analysis revealed higher numbers of mitochondria and secretory follicles, and an increased proliferation rate. Elevated expression levels of IGF1, IGF2, PRLR and ERα were observed in decidualised cells; PRL and ERβ remained mostly below the detection limit, while PGR remained unaffected. The expression of smooth muscle α actin (αSMA), another decidualisation marker, was strongly induced. Among prostaglandin system members, levels of COX2 (PTGS2) and of PGE2-synthase (PTGES) were upregulated. Expression of the PGE2 receptors, PTGER2 and PTGER4, was clearly detectable.
An in vitro decidualisation model with canine uterine stromal cells was successfully established, allowing future, more detailed studies to be undertaken on the underlying molecular and endocrine mechanisms of canine decidualisation.
在家犬（犬属）中，子宫对胚胎存在的反应尚不清楚。胚泡孵化和胚胎着床后开始的紧密的胚胎 - 母体交互作用会导致子宫发生强烈的结构和功能重塑。这个过程的一部分是蜕膜化，包括形态和生化变化，导致母体基质衍生的蜕膜细胞形成。这些细胞是犬胎盘母体的一个组成部分，与母体血管内皮一起是犬内皮绒毛膜胎盘仅有的能够抵抗滋养层侵袭的细胞。这些细胞也是犬胎盘中唯一表达孕酮受体（PGR）的细胞。因此，了解蜕膜化过程对于理解犬类生殖生理学似乎至关重要。
在这里，我们研究了犬子宫基质细胞在体外蜕膜化的能力，从而建立了一个蜕膜化的犬模型。在24 - 72小时的时间进程中采用了dbcAMP介导的方法。使用了来自六只（n = 6）健康、处于发情后期母犬的组织材料（排卵后约2周）。细胞通过鉴别染色进行表征，其中近100%为波形蛋白阳性。应用扫描和透射电子显微镜分析，并用活细胞成像显微镜记录形态变化。研究了几种蜕膜化标志物的表达。
体外培养的基质细胞在与0.5 mM dbcAMP孵育72小时后获得了蜕膜细胞的特征。它们的形状从细长变为圆形，而超微结构分析显示线粒体和分泌小泡数量增加，增殖率提高。在蜕膜化细胞中观察到IGF1、IGF2、PRLR和ERα的表达水平升高；PRL和ERβ大多低于检测限，而PGR不受影响。另一个蜕膜化标志物平滑肌α肌动蛋白（αSMA）的表达被强烈诱导。在前列腺素系统成员中，COX2（PTGS2）和PGE2合酶（PTGES）的水平上调。PGE2受体PTGER2和PTGER4的表达清晰可检测到。
成功建立了一个犬子宫基质细胞的体外蜕膜化模型，这使得未来能够对犬蜕膜化的潜在分子和内分泌机制进行更详细的研究。
