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The Seasonal and Stage-Specific Expression Patterns of HMGB2 Suggest Its Key Role in Spermatogenesis in the Chinese Soft-Shelled Turtle (Pelodiscus sinensis)

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Abstract

HMGB2, a member of the high-mobility group (HMG) proteins, was identified as a male-biased gene and plays a crucial role in the germ cells differentiation of mammals. However, its role in spermatogenesis of turtle is still poorly understood. Here, we cloned the Pelodiscus sinensis HMGB2 and analyzed its expression profile in different tissues and in testis at different developmental ages. P. sinensis HMGB2 mRNA was highly expressed in the testis of 3-year-old turtles (P < 0.01), but was hardly detected in ovaries and other somatic tissues. The results of chemical in situ hybridization (CISH) showed that HMGB2 mRNA was specifically expressed in germ cells, where it was mainly distributed in round spermatids and sperm, but not detected in somatic cells, spermatogonia, primary spermatocytes, or secondary spermatocyte. The relative expression of HMGB2 also responded to seasonal changes in testis development in P. sinensis. In different seasons of the year, the relative expression of HMGB2 transcripts in the testis of 1 year and 2 year olds showed an overall upward trend, whereas, in the testis of 3 year old, it peaked in July and then declined in October. Moreover, in April and July, with an increase in ages, the expression of HMGB2 transcripts showed an upward trend. However, in January and October, there was a decline in expression in testis in 3-year-old turtles. These results showed that HMGB2 is closely related to spermatogenesis in P. sinensis.

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Acknowledgements

This research work was supported by Pearl River S&T Nova Program of Guangzhou (201806010131), National Key R&D Program of China (2018YFD090020), The Project Supported by Guangdong Natural Science Foundation (2018A030313520), Guangdong Agricultural Research System (2019KJ150), Central Public-interest Scientific Institution Basal Research Fund, CAFS (2020TD35, 2020ZJTD01), National Freshwater Genetic Resource Center (NFGR-2020), and International Agricultural Exchange and Cooperation (2130114).

Funding

This study was supported by National Key R&D Program of China [Grant No. 2018YFD090020], The Project Supported by Guangdong Natural Science Foundation [Grant No. 2018A030313520], Guangdong Agricultural Research System [Grant No. 2019KJ150], Central Public-interest Scientific Institution Basal Research Fund, CAFS [Grant Nos. 2020TD35, 2020ZJTD01], National Freshwater Genetic Resource Center [Grant No. NFGR-2020], International Agricultural Exchange and Cooperation [Grant No. 2130114], Pearl River S&T Nova Program of Guangzhou [Grant No. 201806010131].

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Author notes

  1. Wei Li and Junxian Zhu have contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Tropical & Subtropical Fishery Resource Application & Cultivation of Ministry of Agriculture and Rural Affairs, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangdong, Guangzhou, 510380, People’s Republic of China

    Wei Li, Junxian Zhu, Luo Lei, Chen Chen, Xiaoli Liu, Yakun Wang, Xiaoyou Hong, Lingyun Yu, Hongyan Xu & Xinping Zhu

  2. Wuxi Fisheries College, Nanjing Agricultural University, Wuxi, Jiangsu, 214081, People’s Republic of China

    Junxian Zhu, Luo Lei & Xinping Zhu

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  1. Wei Li
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Contributions

XZ, HX, and WL designed the experiments. LL, CC, XL, YW, XH, and LY collected samples. JZ and WL performed the experiments and analyzed the data; JZ wrote the paper. XZ and HX revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Hongyan Xu or Xinping Zhu.

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Supplementary Information

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10528_2022_10229_MOESM1_ESM.tif

Supplementary file1 (TIF 1796 KB)—Supplementary data 1. The sequence homology analysis of cloned P. sinensis HMGB2 cDNA. The underline shows primer binding site.

10528_2022_10229_MOESM2_ESM.tif

Supplementary file2 (TIF 5272 KB)—Supplementary data 2. The HMGB2 mRNA expression in testis by CISH. (a, b and e) No signal was detected by antisense probe. (c and d) Nucleus were stained with PI (red). (a and c) The testis from P. sinensis in 1-year old. (b and d) The testis from P. sinensis in 2-year old. (e) The testis from P. sinensis in 3-year old. Scale bars = 50 µm.

10528_2022_10229_MOESM3_ESM.tif

Supplementary file3 (TIF 224 KB)—Supplementary data 3. The expression of EF1α mRNA in different tissues were analyzed by semi-quantitative PCR.

10528_2022_10229_MOESM4_ESM.tif

Supplementary file4 (TIF 481 KB)—Supplementary data 4. The expression of HMGB2 mRNA in different tissues were analyzed by semi-quantitative PCR.

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Li, W., Zhu, J., Lei, L. et al. The Seasonal and Stage-Specific Expression Patterns of HMGB2 Suggest Its Key Role in Spermatogenesis in the Chinese Soft-Shelled Turtle (Pelodiscus sinensis). Biochem Genet 60, 2489–2502 (2022). https://doi.org/10.1007/s10528-022-10229-0

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