title: Increased accessibility of the N-terminus of testis-specific histone TH2B to antibodies in elongating spermatids.
abstract: Changes in chromatin structure during spermatogenesis were investigated using a monoclonal antibody that immunoreacts with the N-terminus of the testis-specific histone TH2B. This monoclonal antibody, which had been raised against rat tyrosine hydroxylase (TH), cross-reacted with TH2B because of sequence homology at the N-termini of TH and TH2B. The epitope was localized to the N-terminus of TH2B as trypsin-digested chromatin which lacked the N-terminal tail did not react with anti-TH and preincubating anti-TH with a synthetic peptide made from the homologous sequence between TH2B and TH inhibited its binding to TH and TH2B. In histological sections of rat testis, the primary spermatocytes and round spermatids immunoreacted weakly, whereas elongating spermatids at steps 10-12 immunoreacted intensely with anti-TH. Increased staining of elongating spermatids was also observed in mouse and hamster by immunohistochemistry. However, immunoblotting proteins extracted from separated rat testis cells showed no increase in the TH2B content of these late steps of spermatids. The apparent increase in the immunohistochemical staining corresponds to increased accessibility of the epitope in the elongating spermatids. This indicated that the N-terminus of TH2B is less tightly bound to DNA or to other proteins at this time in preparation for the removal of TH2B and other histones.
authors.affiliation: Department of Experimental Radiotherapy, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
authors.forename: E
authors.initials: E
authors.lastname: Unni
authors.forename: A
authors.initials: A
authors.lastname: Mayerhofer
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: Y M
authors.initials: YM
authors.lastname: Bhatnagar
authors.forename: L D
authors.initials: LD
authors.lastname: Russell
authors.forename: M L
authors.initials: ML
authors.lastname: Meistrich
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002843:Chromatin / Q000737:chemistry / Q000235:genetics; D006224:Cricetinae; D003429:Cross Reactions; D000939:Epitopes / Q000235:genetics; D006657:Histones / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008297:Male; D008647:Mesocricetus; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017386:Sequence Homology, Amino Acid; D013087:Spermatids / Q000378:metabolism*; D013091:Spermatogenesis; D013737:Testis / Q000378:metabolism*; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D002843:Chromatin; D000939:Epitopes; D006657:Histones; D009687:Nuclear Proteins; C081018:histone H2B type 1-A; D014446:Tyrosine 3-Monooxygenase