title: Assessment of estradiol-induced gene regulation and proliferation in an immortalized mouse immature Sertoli cell line.
abstract: OBJECTIVE
  The number of Sertoli cells during proliferative phase determines the fate of the germ cells in male reproductive system. A well-characterized cell line may help in better understanding of Sertoli cell biology. Hence, the present study assessed estradiol signaling in a mouse immature Sertoli cell line (MSC-1) as an alternative model in place of primary culture of Sertoli cells.
  
  
  METHODS
  In this study, we used MSC-1 cell line, derived from 10-day old mice. The cell cycle parameters were assessed, and the expression and regulation of Sertoli cell-specific secretory genes (ABP; androgen-binding protein) and tight junction genes (claudin-5, occludin, and vimentin) in response to estradiol was studied.
  
  
  RESULTS
  The results obtained suggested the presence of both estrogen receptors (ERα and ERβ) in MSC-1 cells. In vitro scratch assay and cell-cycle analysis suggested the proliferative effects of estradiol in both time- and dose-dependent manner. The gene expression profiles of ABP, claudin-5, and occludin showed biphasic regulation at low and high doses of estradiol. Analysis of signaling pathways suggested the activation of extracellular signal-regulated kinase (ERK) pathway with significantly increased pERK/ERK ratio (p<0.05). The results also suggested down regulation in the expression of mir-17 family members (mir-17, mir-20b, and mir-106a) (p<0.05).
  
  
  CONCLUSIONS
  Considering the limited number of Sertoli cell lines and long-term survival inability of primary culture of Sertoli cells, MSC-1 cells could be a potential cell line for understanding the mechanisms of various cellular events in Sertoli cells.
authors.affiliation: Molecular Endocrinology Laboratory, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247 667, Uttarakhand, India.
authors.forename: Narender
authors.initials: N
authors.lastname: Kumar
authors.affiliation: Molecular Endocrinology Laboratory, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247 667, Uttarakhand, India.
authors.forename: Swati
authors.initials: S
authors.lastname: Srivastava
authors.affiliation: University Wurzburg, Department of Anaesthesia and Critical Care, Oberduerrbacher Strasse 6, 97080 Wurzburg, Germany.
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Burek
authors.affiliation: University Wurzburg, Department of Anaesthesia and Critical Care, Oberduerrbacher Strasse 6, 97080 Wurzburg, Germany.
authors.forename: Carola Y
authors.initials: CY
authors.lastname: Förster
authors.affiliation: Molecular Endocrinology Laboratory, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247 667, Uttarakhand, India. Electronic address: paroyfbs@iitr.ac.in.
authors.forename: Partha
authors.initials: P
authors.lastname: Roy
mesh_terms: D000818:Animals; D016466:CHO Cells; D002461:Cell Line, Transformed; D049109:Cell Proliferation / Q000187:drug effects / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D006224:Cricetinae; D003412:Cricetulus; D004958:Estradiol / Q000494:pharmacology*; D008297:Male; D051379:Mice; D012708:Sertoli Cells / Q000187:drug effects / Q000502:physiology*; D059467:Transcriptome / Q000187:drug effects / Q000502:physiology*
chemical_list: D004958:Estradiol
keywords: Epigenetic; Estrogen receptors; Estrogens; MAPK pathway; MicroRNAs; Sertoli cell proliferation