OBJECTIVE

To study the mRNA and protein expressions of Dnajb13 and its localization in the testis of the mouse with cryptorchidism and its association with the apoptosis of spermatogenic cells.

METHODS

The localization of Dnajb13 in the spermatogenic cells of 8-week-old mice was detected by immunohistochemistry. The model of unilateral cryptorchidism was surgically established in the mice and verified by TUNEL, flow cytometry and morphological observation. The apoptosis of the spermatogenic cells was analyzed and the mRNA and protein expressions of Dnajb13 in both cryptorchid and healthy testes were determined by quantitative polymerase chain reaction (qPCR), Western blot and immunohistochemistry at 1, 2, 3, 4, 5, 6, 9 and 15 days after modeling.

RESULTS

Immunohistochemistry showed that Dnajb13 was localized in the elongated spermatids at steps 9－16 of spermiogenesis in the testis tissue of the healthy mice. TUNEL and flow cytometry manifested that the round spermatids at step 1 and primary spermatocytes in miosis were most sensitive to elevated temperature. After modeling, apoptosis was first observed in the round spermatids at steps 1－8, which were decreased from 17.09% to 6.52% (P < 0.05), then in the spermatids during metamorphosis at steps 9－16, and then in the primary spermatocytes. At 3 days after surgery, the expression of Dnajb13 mRNA in the cryptorchid testis was 1.6 times higher than that in the healthy one (P < 0.05) and decreased at 4 days, 1.2 times that of the normal. The expression of the Dnajb13 protein exhibited no significant change at 1－3 days, but a 0.68-fold reduction at 4 days (P < 0.05) and a 0.4-fold reduction at 9 days. Immunohistochemical staining revealed the expression of the Dnajb13 protein in the apoptotic multinucleated giant cells at 6 days.

CONCLUSIONS

Dnajb13 is localized in the spermatids during metamorphosis and in the tails of mature sperm in adult mice, involved in sperm metamorphosis and sperm flagellum formation, and expressed in apoptotic multinucleated giant cells in the cryptorchid testis, which may be associated with the apoptosis of round spermatids at stages Ⅵ－Ⅷ.