Characterization of Newly Established Clonal Oviductal Cell Lines and Differential Hormonal Regulation of Gene Expression

Abstract

Oviductal functions have been studied mainly in primary epithelial cell culture and organ culture. However, secretory cells and ciliated cells coexist in the epithelium, and the small size of the oviduct limits the sources of both epithelial and stromal cells. To circumvent the limits, we attempted to establish clonal cell lines from an oviduct of a p53-deficient mouse. An oviduct was enzymatically digested and cultured in medium containing 10% fetal calf serum supplemented with estradiol-17beta. Morphologically distinct clones (10 epithelial and 4 fibroblastic clones) were established, and all clones expressed estrogen receptor alpha and progesterone receptor. Expression of a mouse oviduct-specific glycoprotein gene as a marker of secretory cells was limited in one clone and was stimulated by estrogens and suppressed by progesterone. Expression of helix factor hepatocyte nuclear factor/forkhead homologue-4 gene as a marker of ciliated cells was limited in two clones and was suppressed by estrogens. The two genes were never coexpressed in any clones. The results strongly suggest that the oviductal epithelium consists of two functionally determined populations. To our knowledge, this is the first establishment of functional clonal cell lines of the oviduct and makes it possible to study independently two oviductal functions, secretion and ciliogenesis.

Follistatin-like-1, a diffusible mesenchymal factor determines the fate of epithelium

“…Ovgp1 is one of the secretory proteins of the mouse oviduct (32,33), and Foxj1 is a transcription factor specifically involved in ciliogenesis (34)(35)(36). 6B, a cilial epithelial cell line, and AB, a secretory epithelial cell line, which were established from adult oviducts in our previous study (20), expressed Foxj1 and Ovgp1, respectively ( Fig. 1 A and B).…”

“…The major difficulties in analyzing the mechanisms of organogenesis are from the nature of the developmental events that occur three-dimensionally during a brief period in tissues of small scale. To circumvent these difficulties, we have established oviductal epithelial clonal cell lines from p53 −/− mice (38) in the present and previous studies (20). These epithelial cell lines allowed us to develop model systems in which we can analyze the oviductal development in vitro and in vivo.…”

“…In adult mesenchyme, such determining cells disappear (Fstl1-secreting cells were undetected). Instead, the mesenchyme has enhancer cells that enhance the expression of Ovgp1 and Foxj1 in secretory epithelial cells and in cilial epithelial cells, respectively (20). Dual origin of mesenchymal tissues is reported in mouse mammary gland embryogenesis (42).…”

“…Our previous study suggested that the oviduct contains at least two distinct epithelial cell populations rather than a single population with a transitional phenotype (20,21). How are their fates determined?…”

“…To circumvent problems of small tissue scale and to develop reproducible in vitro models, we established epithelial and mesenchymal clonal cell lines and attempted to investigate the mechanism of how mesenchyme determines the fate of epithelial cells in the oviduct. We have already demonstrated that the p53 −/− mouse is a useful source for establishing clonal cell lines with tissue-specific phenotypes (20,(22)(23)(24)(25) and developmental-stage-specific phenotypes (21,(26)(27)(28)(29).…”