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Evaluation of apoptotic genes expression and its protein after treatment of cryptorchid mice
Forouzan Absalan1,2, Mansoureh Movahedin1, Seyed Javad Mowla3
1Dept. of Anatomical Sciences, School of Medical Sciences, Tarbiat Modarres University, Tehran, Iran.
Introduction:
Cryptorchidism has been proved to cause apoptosis in germ cells in respond to changes in the stimulation levels of specific physiological events. The purpose of this study was to determine whether treatment of bilateral cryptorchidism was associated with alterations in testicular gene expression.
Methods:
To induce bilateral cryptorchid model, immature mice were anesthetized and a small incision was made in the abdominal skin and peritoneum, then fat pad at the upper end of testis was sutured to the peritoneum. Transcript level of Bax, Bcl-2 proper, p53 and survivin mRNA and protein were determined after performing the two treatment methods: surgical return of testis into scrotum (Exp1) and transplantation of spermatogonial stem cells with later orchidopexy (Exp2), performed 2 and 3 months after heat exposure, respectively.
Results:
RT-PCR data showed decreased levels of p53 and Bax expression as well as decreased levels of Bcl-2 mRNA in treatment groups especially after transplantation compared with control group. The expression of survivin 140 was increased significantly after treatment, whereas that of survivin 40 was lower especially in the orchidopexy group. Immunohistochemistry staining showed that the intensity of Bax expression mainly was decreased in treated cryptorchid testis and rates of Bcl-2 were increased significantly, but expression of p53 and survivin proteins did not changed significantly after treatment.
Discussion:
These observations suggest that cell-type-specific and many apoptotic systems control germ cell apoptosis after treatment of cryptorchidism.
Insights
Treatment for cryptorchidism (undescended testes) in mice altered gene expression related to germ cell apoptosis. These findings suggest complex cellular mechanisms control germ cell survival after treatment.
Area of Science:
- Reproductive Biology
- Molecular Biology
- Cell Biology
Background:
- Cryptorchidism is known to induce germ cell apoptosis due to altered physiological stimulation.
- Understanding gene expression changes post-treatment is crucial for reproductive health.
Purpose of the Study:
- To investigate alterations in testicular gene expression following treatment for bilateral cryptorchidism.
- To compare the effects of surgical correction versus stem cell transplantation on gene expression.
Main Methods:
- A bilateral cryptorchidism mouse model was established.
- Gene and protein expression of Bax, Bcl-2, p53, and survivin were analyzed after surgical orchidopexy or spermatogonial stem cell transplantation.
- RT-PCR and immunohistochemistry were employed for analysis.
Main Results:
- Treatment led to decreased p53 and Bax mRNA, and decreased Bcl-2 mRNA.
- Survivin 140 mRNA increased, while Survivin 40 mRNA decreased post-treatment.
- Immunohistochemistry revealed decreased Bax and increased Bcl-2 protein intensity, with no significant change in p53 or survivin proteins.
Conclusions:
- Germ cell apoptosis following cryptorchidism treatment is regulated by cell-type-specific apoptotic systems.
- Treatment interventions modulate key genes involved in apoptosis, impacting germ cell fate.
