Stadnicka Katarzyna, Sławińska Anna, Dunisławska Aleksandra, Pain Bertrand, Bednarczyk Marek
Department of Animal Biochemistry and Biotechnology, UTP University of Science and Technology, Mazowiecka 28, 85-084, Bydgoszcz, Poland.
University of Lyon, Université Lyon 1, INSERM, INRA, Stem Cell and Brain Research Institute, U1208, USC1361, Bron, France.
BMC Dev Biol. 2018 Apr 4;18(1):9. doi: 10.1186/s12861-018-0168-2.
In this work we have determined molecular signatures of oviduct epithelial and progenitor cells. We have proposed a panel of selected marker genes, which correspond with the phenotype of oviduct cells of a laying hen (Gallus gallus domesticus) and quail (Coturnix japonica). We demonstrated differences in characteristics of those cells, in tissue and in vitro, with respect to different anatomical and functional parts of the oviduct (infundibulum (INF), distal magnum (DM, and proximal magnum (PM)). The following gene expression signatures were studied: (1) oviduct markers (estrogen receptor 1, ovalbumin, and SPINK7 - ovomucoid), (2) epithelial markers (keratin 5, keratin 14, and occludin) and (3) stem-like/progenitor markers (CD44 glycoprotein, LGR5, Musashi-1, and sex determining region Y-box 9, Nanog homebox, OCT4/cPOUV gene encoding transcription factor POU5F3).
In chicken, the expression of oviduct markers increased toward the proximal oviduct. Epithelial markers keratin14 and occludin were high in distal oviduct and decreased toward the proximal magnum. In quail oviduct tissue, the gene expression pattern of oviduct/epithelial markers was similar to chicken. The markers of progenitors/stemness in hen oviduct (Musashi-1 and CD44 glycoprotein) had the highest relative expression in the infundibulum and decreased toward the proximal magnum. In quail, we found significant expression of four progenitor markers (LGR5 gene, SRY sex determining region Y-box 9, OCT4/cPOUV gene, and CD44 glycoprotein) that were largely present in the distal oviduct. After in vitro culture of oviduct cells, the gene expression pattern has changed. High secretive potential of magnum-derived cells diminished by using decreased abundance of mRNA. On the other hand, chicken oviduct cells originating from the infundibulum gained ability to express OVM and OVAL. Epithelial character of the cells was maintained in vitro. Among progenitor markers, both hen and quail cells expressed high level of SOX9, LGR5 and Musashi-1.
Analysis of tissue material revealed gradual increase/decrease pattern in majority of the oviduct markers in both species. This pattern changed after the oviductal cells have been cultured in vitro. The results can provide molecular tools to validate the phenotype of in vitro biological models from reproductive tissue.
在本研究中，我们确定了输卵管上皮细胞和祖细胞的分子特征。我们提出了一组选定的标记基因，它们与蛋鸡（家鸡）和鹌鹑输卵管细胞的表型相对应。我们展示了这些细胞在组织和体外，相对于输卵管不同解剖和功能部位（漏斗部（INF）、远端峡部（DM）和近端峡部（PM））的特征差异。研究了以下基因表达特征：（1）输卵管标记物（雌激素受体1、卵清蛋白和SPINK7 - 卵类粘蛋白），（2）上皮标记物（角蛋白5、角蛋白14和闭合蛋白），以及（3）干细胞样/祖细胞标记物（CD44糖蛋白、LGR5、神经母细胞瘤致癌基因同源物1和性别决定区Y框9、Nanog同源盒、编码转录因子POU5F3的OCT4/cPOUV基因）。
在鸡中，输卵管标记物的表达向输卵管近端增加。上皮标记物角蛋白14和闭合蛋白在输卵管远端较高，向近端峡部降低。在鹌鹑输卵管组织中，输卵管/上皮标记物的基因表达模式与鸡相似。母鸡输卵管中祖细胞/干性的标记物（神经母细胞瘤致癌基因同源物1和CD44糖蛋白）在漏斗部相对表达最高，向近端峡部降低。在鹌鹑中，我们发现四个祖细胞标记物（LGR5基因、SRY性别决定区Y框9、OCT4/cPOUV基因和CD44糖蛋白）有显著表达，它们主要存在于输卵管远端。输卵管细胞体外培养后，基因表达模式发生了变化。峡部来源细胞的高分泌潜能因mRNA丰度降低而减弱。另一方面，来自漏斗部的鸡输卵管细胞获得了表达OVM和OVAL的能力。细胞的上皮特征在体外得以维持。在祖细胞标记物中，母鸡和鹌鹑细胞均高表达SOX9、LGR5和神经母细胞瘤致癌基因同源物1。
对组织材料的分析揭示了两种物种中大多数输卵管标记物呈逐渐增加/减少的模式。输卵管细胞体外培养后这种模式发生了变化。这些结果可为验证生殖组织体外生物学模型的表型提供分子工具。
