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Updated: Aug 11, 2026
Step-specific Sorting of Mouse Spermatids by Flow Cytometry
Published on: December 31, 2015
Temperature sensitivity of primary spermatocyte DNA synthesis in immature mice confirmed by bromodeoxyuridine
B Zhou1, J M Hutson, S Hasthorpe
1F. Douglas Stephens Surgical Research Laboratory, Royal Children's Hospital Research Institute, Melbourne, Australia.
Objective:
To investigate the relationship between temperature and DNA synthesis of immature germ cells and to determine whether the early primary spermatocytes proliferate at a 'scrotal' temperature of 32 degrees C in vitro.
Materials And Methods:
Day-7 mouse testes (n = 16) were cultured with 10% fetal calf serum (FCS) for 3 days at 32 degrees C or 37 degrees C and labelled by bromodeoxyuridine (BrDU) for a further hour. The BrDU-labelled cells were detected by immunohistochemical staining using a monoclonal anti-BrDU antibody. The numbers of primary spermatocytes with BrDU-labelling or unlabelled in each tubule were determined as an index of spermatocyte DNA synthesis. In addition, the cultured media at different temperatures were collected and the testosterone levels measured by radioimmunoassay.
Results:
The numbers of primary spermatocytes (P < 0.01) and the BrDU-labelling index in primary spermatocytes per tubule at 32 degrees C in vitro were significantly higher (P < 0.001) than in the culture at 37 degrees C. The testosterone levels in the culture media at 32 degrees C were also markedly higher than in the culture at 37 degrees C (P < 0.01).
Conclusion:
These observations indicate that DNA synthesis of early primary spermatocytes and testosterone production can be stimulated by lower testicular temperature, even in immature mouse testes that are naturally located in the intra-abdominal cavity.
