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Genetic and histopathological analysis of spermatogenesis after short-term testicular torsion in rats

Pediatric Research (2023)Cite this article

Abstract

Background

Patients with testicular torsion (TT) may exhibit impaired spermatogenesis from reperfusion injury after detorsion surgery. Alteration in the expressions of spermatogenesis-related genes induced by TT have not been fully elucidated.

Methods

Eight-week-old Sprague-Dawley rats were grouped as follows: group 1 (sham-operated), group 2 (TT without reperfusion) and group 3 (TT with reperfusion). TT was induced by rotating the left testis 720° for 1 h. Testicular reperfusion proceeded for 24 h. Histopathological examination, oxidative stress biomarker measurements, RNA sequencing and RT-PCR were performed.

Results

Testicular ischemia/reperfusion injury induced marked histopathological changes. Germ cell apoptosis was significantly increased in group 3 compared with group 1 and 2 (mean apoptotic index: 26.22 vs. 0.64 and 0.56; p = 0.024, and p = 0.024, respectively). Johnsen score in group 3 was smaller than that in group 1 and 2 (mean: 8.81 vs 9.45 and 9.47 points/tubule; p = 0.001, p < 0.001, respectively). Testicular ischemia/reperfusion injury significantly upregulated the expression of genes associated with apoptosis and antioxidant enzymes and significantly downregulated the expression of genes associated with spermatogenesis.

Conclusion

One hour of TT followed by reperfusion injury caused histopathological testicular damage. The relatively high Johnsen score indicated spermatogenesis was maintained. Genes associated with spermatogenesis were downregulated in the TT rat model.

Impact

  • How ischemia/reperfusion injury in testicular torsion (TT) affects the expressions of genes associated with spermatogenesis has not been fully elucidated.

  • This is the first study to report comprehensive gene expression profiles using next generation sequencing for an animal model of TT.

  • Our results revealed that ischemia/reperfusion injury downregulated the expression of genes associated with spermatogenesis and sperm function in addition to histopathological damage, even though the duration of ischemia was short.

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Fig. 1: Photomicrographs of testicular tissues stained with hematoxylin and eosin.
Fig. 2: TUNEL staining of testicular tissues.
Fig. 3: Gene expression profiling of testis tissue.
Fig. 4: Oxidative stress biomarkers in testis tissue.

Data availability

The RNA-seq dataset generated during and/or analyzed during the current study is available in the DDBJ Sequenced Read Archive (DRA) under the accession number DRA015199.

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Acknowledgements

Expert technical assistance was provided by Machi Kawauchi and Kazuya Tanaka. We thank Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript.

Funding

This work was supported by JSPS KAKENHI Grant Numbers JP16H06276 (AdAMS) and JP19K18355.

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Authors and Affiliations

  1. Department of Pediatric Surgery, Faculty of Medicine, Kagawa University, Miki-cho, Kagawa Prefecture, Japan

    Hiroto Katami, Takayuki Fujii, Aya Tanaka & Ryuichi Shimono

  2. Department of Anatomy and Neurobiology, Faculty of Medicine, Kagawa University, Miki-cho, Kagawa Prefecture, Japan

    Shingo Suzuki, Ken-ichi Ohta & Takanori Miki

  3. Department of Pathology and Host Defense, Faculty of Medicine, Kagawa University, Miki-cho, Kagawa Prefecture, Japan

    Masaki Ueno

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  1. Hiroto Katami
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Contributions

Substantial contributions to conception and design, acquisition of data, or analysis and interpretation of data: H.K., R.S., S.S., M.U., T.M., T.F., A.T., and K.O. Drafting the article or revising it critically for important intellectual content: H.K., S.S., M.U., and R.S. Final approval of the version to be published: H.K., S.S., T.F., M.U., A.T., K.O., T.M., and R.S.

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Correspondence to Ryuichi Shimono.

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Katami, H., Suzuki, S., Fujii, T. et al. Genetic and histopathological analysis of spermatogenesis after short-term testicular torsion in rats. Pediatr Res (2023). https://doi.org/10.1038/s41390-023-02638-7

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