Immunohistochemical study of seminiferous epithelium in adult bovine testis using monoclonal antibodies against Ki-67 protein and proliferating cell nuclear antigen (PCNA)

Cell Tissue Res. 1996 Feb;283(2):191-201. doi: 10.1007/s004410050529.


The distribution pattern of proliferating cell nuclear antigen (PCNA) and Ki-67 protein was studied in adult bovine seminiferous epithelium by means of immunohistochemistry using monoclonal antibodies. Tailoring the methodological protocol for each of the two proliferation markers was a necessary prerequisite for obtaining optimal results in tubular sections and whole-mounts. A-, I- and B-spermatogonia displayed PCNA-positive nuclei, except during meta-, ana- and telophases of mitosis. PCNA-negative nuclei in the basal tubular compartment, excluding those from non-cycling Sertoli cells, belonged to the spermatogonia precursor cell line. However, only about 30%, 45% and 47% of the respective A-, I-, B-spermatogonia had positive nuclei after exposure to the MIB-1 antibody directed against the Ki-67 protein. Spermatogonia with MIB-1-negative nuclei represented cells in the G1-phase. Both antibodies reacted intensely with the nuclei of preleptotene primary spermatocytes. PCNA reactivity was also present during leptotene through pachytene. Ki-67 protein expression was absent during leptotene and zygotene but was again encountered during pachytene and meiosis I and II. Anti-PCNA/anti-protein gene product 9.5 double-labelling indicated that the transition from spermatogonia precursor cells into A1-spermatogonia is not strictly synchronized in a given tubular segment, a possible reason for the flexibility in A-spermatogonial propagation seen in bovine seminiferous tubules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Biomarkers
  • Cattle
  • Cell Cycle
  • Cell Division
  • Epithelial Cells
  • Epithelium / chemistry
  • Ki-67 Antigen
  • Male
  • Meiosis
  • Neoplasm Proteins / analysis*
  • Neoplasm Proteins / immunology
  • Nuclear Proteins / analysis*
  • Nuclear Proteins / immunology
  • Proliferating Cell Nuclear Antigen / analysis*
  • Proliferating Cell Nuclear Antigen / immunology
  • Seminiferous Tubules / chemistry
  • Seminiferous Tubules / cytology*
  • Spermatogenesis
  • Spermatogonia / chemistry


  • Antibodies, Monoclonal
  • Biomarkers
  • Ki-67 Antigen
  • Neoplasm Proteins
  • Nuclear Proteins
  • Proliferating Cell Nuclear Antigen