Proteomic analysis of androgen-regulated protein expression in a mouse fetal vas deferens cell line.
During sex differentiation, androgens are essential for development of the male genital tract. The Wolffian duct is an androgen-sensitive target tissue that develops into the epididymis, vas deferens, and seminal vesicle. The present study aimed to identify androgen-regulated proteins that are involved in development of Wolffian duct-derived structures. We have used male mouse embryos transgenic for temperature-sensitive simian virus 40 large tumor antigen at 18 d of gestation, to generate immortalized mouse fetal vas deferens (MFVD) parental and clonal cell lines. The MFVD parental and clonal cell lines express androgen receptor protein and show features of Wolffian duct mesenchymal cells. Clonal cell line MFVD A6 was selected for proteomic analysis and cultured in the absence or presence of androgens. Subsequently, two-dimensional gel electrophoresis was performed on total cell lysates. Differentially expressed proteins were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and two androgen-regulated proteins were identified as mElfin and CArG-binding factor-A (CBF-A). CBF-A and mElfin are known to bind to cytoskeletal F-actin. Both proteins appeared to be regulated by androgens at the posttranslational level, possibly involving phosphorylation. Posttranslational modification of mElfin and CBF-A by androgens may be associated with a cytoskeletal change that is involved in androgen-regulated gene expression.
Umar
Arzu
A
Department of Reproduction & Development, Erasmus MC, University Medical Center Rotterdam, The Netherlands. a.umar@erasmusmc.nl
Luider
Theo M
TM
Berrevoets
Cor A
CA
Grootegoed
J Anton
JA
Brinkmann
Albert O
AO
D000728:Androgens / Q000494:pharmacology*; D000818:Animals; D002461:Cell Line, Transformed / Q000737:chemistry / Q000502:physiology; D019070:Cell Lineage / Q000502:physiology; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D060588:LIM Domain Proteins; D008297:Male; D051379:Mice; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D011247:Pregnancy; D040901:Proteomics; D011944:Receptors, Androgen / Q000032:analysis* / Q000235:genetics; D012097:Repressor Proteins / Q000032:analysis / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D017154:Stromal Cells / Q000166:cytology; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics; D014649:Vas Deferens / Q000166:cytology / Q000196:embryology* / Q000502:physiology*
D000728:Androgens; D004268:DNA-Binding Proteins; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D060588:LIM Domain Proteins; C106649:Ldb2 protein, mouse; D011944:Receptors, Androgen; D012097:Repressor Proteins; D014157:Transcription Factors; C077048:Hnrnpab protein, mouse