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Published on: February 6, 2018
[NEK2 gene expression in mouse cryptorchidism model and its mechanism involved in apoptosis]
1Department of Pediatric Surgery, Affiliated Hospital of Nantong University, Nantong 226000, China.
Abstract:
Objective: To observe the expression of NEK2 mRNA and protein in the cryptorchidism mice model, and to explore its role in apoptosis of testicular tissue. Methods: A mouse cryptorchid model was constructed, and the spermatids in the spermatic tubules were observed by HE staining. Apoptosis was detected by Tunel test, and expression of NEK2 mRNA and protein was detected by RT-PCR and immunohistochemistry, respectively. Results: After the mouse cryptorchidism model was successfully constructed, the HE staining results showed that the damage of spermatogonia cells, primary spermatocytes and sperm cells in the seminiferous tubules became more severe with time. The results of Tunel test showed that the number of apoptotic cells first increased and then decreased, 1, 3, 6, 9 and 15 d apoptotic cells were 3.67±2.08 (t=2, P=0.0412), 7.67±1.53 (t=6.325, P=0.003), 17.67±3.51 (t=7.906, P=0.001), 30.67±3.51 (t=14.072, P<0.001) and 14.33±3.21 (t=6.860, P=0.002). The results of immunohistochemistry showed that NEK2 protein was expressed in the nucleus and cytoplasm in normal testis and cryptorchidism. RT-PCR and immunohistochemistry showed that expression of NEK2 mRNA and protein gradually increased after modeling. After reaching the peak, the expression gradually decreased with time, and was significantly lower than the normal control group. Conclusion: The trend of NEK2 expression in cryptorchidism tissue is consistent with the trend of cell apoptosis in cryptorchidism tissue, suggesting that abnormal expression of NEK2 may affect the damage of sperm cells in the seminiferous tubules through apoptosis, leading to infertility in patients with cryptorchidism.
Insights
Abnormal NEK2 expression in cryptorchidism mice correlates with testicular apoptosis, suggesting a role in infertility. This study observed NEK2 mRNA and protein levels alongside cell death in a mouse model of undescended testes.
Area of Science:
- Reproductive Biology
- Molecular Biology
- Cell Biology
Background:
- Cryptorchidism, or undescended testes, is a common congenital condition associated with male infertility.
- The underlying molecular mechanisms contributing to testicular damage in cryptorchidism require further elucidation.
- NEK2 (NIMA-related kinase 2) is a cell cycle-regulated kinase implicated in various cellular processes.
Purpose of the Study:
- To investigate the expression patterns of NEK2 mRNA and protein in a mouse model of cryptorchidism.
- To explore the potential role of NEK2 in testicular apoptosis within this model.
- To correlate NEK2 expression with the observed damage in seminiferous tubules.
Main Methods:
- Construction of a mouse model for cryptorchidism.
- Histological examination of testicular tissue using Hematoxylin and Eosin (HE) staining.
- Assessment of apoptosis via Terminal deoxynucleotidyl transferase dUTP nick end labeling (Tunel) assay.
- Quantification of NEK2 mRNA and protein levels using Reverse Transcription Polymerase Chain Reaction (RT-PCR) and immunohistochemistry, respectively.
Main Results:
- HE staining revealed progressive damage to spermatogonia, primary spermatocytes, and sperm cells over time in the cryptorchidism model.
- Tunel assay indicated a biphasic pattern of apoptosis, initially increasing and then decreasing, with a significant peak observed.
- NEK2 mRNA and protein expression levels demonstrated a temporal correlation with the observed apoptosis, increasing after modeling, peaking, and subsequently declining.
Conclusions:
- The expression trend of NEK2 in cryptorchidism tissue mirrors the pattern of cellular apoptosis.
- Abnormal NEK2 expression is implicated in the pathogenesis of testicular damage in cryptorchidism, potentially through the induction of apoptosis.
- These findings suggest NEK2 as a potential factor contributing to infertility in patients with cryptorchidism.
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The Extrinsic Apoptotic Pathway

