title: Expression analysis of RNA-binding motif gene on Y chromosome (RBMY) protein isoforms in testis tissue and a testicular germ cell cancer-derived cell line (NT2).
abstract: RNA-binding motif gene on Y chromosome (RBMY), a germ cell-specific nuclear protein, is known as a key factor in spermatogenesis and disorders associated with this protein have been recognized to be related to male infertility. Although it was suggested that this protein could have different functions during germ cell development, no studies have been conducted to uncover the mechanism of this potential function yet. Here, we analyzed the expression pattern of RBMY protein isoforms in testis compared to NT2, a testicular germ cell cancer-derived cell line, to test probability of differential expression of RBMY protein isoforms at different spermatogenesis stages.
  
  
  
  Full length and a segment of RBMY gene were cloned and expressed in E. coli. Anti-human RBMY antibody was produced in rabbit using the recombinant proteins as antigen. Western-blot and immunofluorescence were conducted for detection and comparison of RBMY protein isoforms.
  
  
  
  Selected segment of RBMY protein resulted in producing a mono-specific antibody. As results shows, only the longest isoform of RBMY was expressed at protein level in NT2 cell line, while three isoforms of this protein were detected in the whole testis lysate.
  
  
  
  The results imply that different alternative splicing may happen in testis cells and probably difference of RBMY function during spermatogenesis is due to the differential expression of RBMY protein isoforms. These results and further experiments on RBMY isoforms can help to obtain a better understanding of the function of this protein, which may increase our knowledge about spermatogenesis and causes of male infertility.
authors.affiliation: Dept. of Molecular Systems Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Alikhani
authors.affiliation: Dept. of Molecular Systems Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Sharifi Tabar
authors.affiliation: Dept. of Molecular Systems Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
authors.forename: Shahab
authors.initials: S
authors.lastname: Mirshahvaladi
authors.affiliation: Animal Core Facility of Reproductive Biomedicine Research Center, 
  Royan Institute for Biotechnology, ACECR, Tehran, Iran.
authors.forename: Abolfazl
authors.initials: A
authors.lastname: Kheimeh
authors.affiliation: Dept. of Andrology at Reproductive Biomedicine Research Center, Royan Institute, ACECR, Tehran, Iran.
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Sadighi Gilani
authors.affiliation: Dept. of Andrology at Reproductive Biomedicine Research Center, Royan Institute, ACECR, Tehran, Iran.
authors.forename: Marjan
authors.initials: M
authors.lastname: Sabbaghian
mesh_terms: D000906:Antibodies / Q000378:metabolism; D015153:Blotting, Western; D045744:Cell Line, Tumor; D041322:Chromosomes, Human, Y / Q000378:metabolism*; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006801:Humans; D008297:Male; D009373:Neoplasms, Germ Cell and Embryonal / Q000378:metabolism* / Q000473:pathology; D009687:Nuclear Proteins / Q000276:immunology / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000276:immunology / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis; D015203:Reproducibility of Results; D013736:Testicular Neoplasms / Q000378:metabolism* / Q000473:pathology; D013737:Testis / Q000378:metabolism* / Q000473:pathology
chemical_list: D000906:Antibodies; D003062:Codon; D009687:Nuclear Proteins; D020033:Protein Isoforms; C501499:RBMY1A1 protein, human; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins
keywords: Protein isoforms; Spermatogenesis; Male infertility