Alterations in chromosomal synapses and DNA repair in apoptotic spermatocytes of Mus m. domesticus

Eur J Histochem. 2016 Jun 14;60(2):2677. doi: 10.4081/ejh.2016.2677.

Abstract

We investigated whether apoptotic spermatocytes from the mouse Mus m. domesticus presented alterations in chromosomal synapses and DNA repair. To enrich for apoptotic spermatocytes, the scrotum's temperature was raised by partially exposing animals for 15 min to a 42ºC water bath. Spermatocytes in initial apoptosis were identified in situ by detecting activated Caspase-9. SYCP1 and SYCP3 were markers for evaluating synapses or the structure of synaptonemal complexes and Rad51 and γH2AX for detecting DNA repair and chromatin remodeling. Apoptotic spermatocytes were concentrated in spermatogenic cycle stages III-IV (50.3%), XI-XII (44.1%) and IX-X (4.2%). Among apoptotic spermatocytes, 48% were in middle pachytene, 44% in metaphase and 6% in diplotene. Moreover, apoptotic spermatocytes showed several structural anomalies in autosomal bivalents, including splitting of chromosomal axes and partial asynapses between homologous chromosomes. gH2AX and Rad51 were atypically distributed during pachytene and as late as diplotene and associated with asynaptic chromatin, single chromosome axes or discontinuous chromosome axes. Among apoptotic spermatocytes at pachytene, 70% showed changes in the structure of synapses, 67% showed changes in gH2AX and Rad51 distribution and 50% shared alterations in both synapses and DNA repair. Our results showed that apoptotic spermatocytes from Mus m. domesticus contain a high frequency of alterations in chromosomal synapses and in the recruitment and distribution of DNA repair proteins. Together, these observations suggest that these alterations may have been detected by meiotic checkpoints triggering apoptosis.

MeSH terms

  • Animals
  • Apoptosis*
  • Caspase 9 / metabolism
  • Cell Cycle Proteins
  • Cell Cycle*
  • Chromosome Pairing*
  • DNA Repair*
  • DNA-Binding Proteins
  • Male
  • Mice
  • Nuclear Proteins / metabolism
  • Rad51 Recombinase / metabolism
  • Spermatocytes / metabolism*
  • Spermatocytes / pathology

Substances

  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • Nuclear Proteins
  • Sycp1 protein, mouse
  • Sycp3 protein, mouse
  • Rad51 Recombinase
  • Rad51 protein, mouse
  • Casp9 protein, mouse
  • Caspase 9

Grants and funding

Funding: This work was supported by FONDECYT Project #1120160 and VID Universidad de Chile, and Project CGL2014-53106-P from Ministerio de Economía y Competitividad (Spain).