Walter I, Miller I
Institute of Histology and Embryology, Veterinary University Vienna, Austria.
Histochem J. 1996 Oct;28(10):671-80. doi: 10.1007/BF02409004.
Cultures of bovine oviduct epithelial cells are widely used in co-culture systems to improve the results of in vitro fertilization. The aim of the present study was to evaluate the suitability of S-100 protein as a differentiation marker for bovine oviduct epithelial cells in vitro. The distribution of S-100 alpha and S-100 beta was examined immunohistochemically in bovine oviduct epithelium in situ and in primary cell cultures derived from it. Three segments of the Fallopian tube (isthmus, ampulla and fimbriae) were compared and analysed during different stages of the oestrus cycle (luteal phase and follicular phase). Ciliated and non-ciliated cells of the epithelium reacted with anti-S-100 alpha, S-100a (alpha beta) and S-100 beta antibodies, except for isthmic non-ciliated cells, which did not bind anti-S-100 beta or anti-S-100a (alpha beta). In addition, basal cells never showed immunoreactivity for S-100. In confluent monolayers of cultured oviduct epithelial cells, disappearance of reactivity for S-100 paralleled morphological signs of dedifferentiation (loss of cilia, cytoplasmic vacuolization). Free-floating oviduct epithelial cells, in contrast, retained morphological differentiation and still expressed S-100 antigen even after seven days in vitro. The immunohistochemical findings were confirmed by polyacrylamide gel electrophoresis and Western blotting. The results indicate that the presence of S-100 is closely connected to morphological differentiation and to the specific functional condition of bovine oviduct epithelial cells.
牛输卵管上皮细胞培养物广泛应用于共培养系统,以提高体外受精的成功率。本研究旨在评估S-100蛋白作为牛输卵管上皮细胞体外分化标志物的适用性。采用免疫组织化学方法检测S-100α和S-100β在牛输卵管原位上皮及原代细胞培养物中的分布。比较并分析了发情周期不同阶段(黄体期和卵泡期)输卵管的三个节段(峡部、壶腹部和伞部)。上皮的纤毛细胞和非纤毛细胞与抗S-100α、S-100a(αβ)和抗S-100β抗体发生反应,但峡部非纤毛细胞不与抗S-100β或抗S-100a(αβ)结合。此外,基底细胞对S-100从未表现出免疫反应性。在培养的输卵管上皮细胞汇合单层中,S-100反应性的消失与去分化的形态学特征(纤毛丧失、细胞质空泡化)平行。相比之下,游离的输卵管上皮细胞即使在体外培养7天后仍保持形态分化并仍表达S-100抗原。聚丙烯酰胺凝胶电泳和蛋白质印迹法证实了免疫组织化学结果。结果表明,S-100的存在与牛输卵管上皮细胞的形态分化及特定功能状态密切相关。