S-100 protein subunits in bovine oviduct epithelium: in situ distribution and changes during primary cell culture

Abstract

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.