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Profiling of testis-specific or testis-predominant genes expressed in mouse male germ cell lines GC-1 and GC-2

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Abstract

Background

Spermatogenesis is a tightly organized process that utilizes an intrinsic genetic program composed of germ cell-specific genes. Although mouse germ cell-related cell lines are available, few germ cell-specific genes have been comprehensively identified in such cell lines.

Objective

We aimed to profile gene expression in the male mouse germ cell-related cell lines, GC-1 and GC-2, characterize their transcriptomic nature, and identify potential testis- or germ cell-specific or -predominant genes expressed in these cell lines.

Methods

We performed profiling analysis of genes transcribed in the mouse germ cell-related cell lines, GC-1 and GC-2, using our previous microarray data together with public transcriptome information. We analyzed the expression of a number of the cell line genes predicted to be preferentially expressed in testis by RT-PCR.

Results

We found that most testis-specific or -predominant mRNAs are not expressed in GC-1 and GC-2 cells, implying that these cell lines have lost their testis- or germ cell-specific genetic characteristics. RT-PCR analysis of genes predicted to be expressed in the cell lines with preferential testicular expression showed the testis-specific or -predominant expression of nine genes and verified four of them as being expressed in the germ cell lines. Among them, only cyclin-dependent kinase inhibitor 3 genes (Cdkn3) showed testis and germ cell specificity.

Conclusion

Our study provides extensive transcriptomic information to shed light on the limited testicular characteristics of the mouse male germ cell-derived cell lines, GC-1 and GC-2, and offers a list of germ cell line genes with testicular preference.

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Data availability

The data supporting this study’s findings are available from the corresponding author upon reasonable request.

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Funding

This was supported by the National Research Foundation of Korea (NRF) grant (2022R1A2C1003441), and GIST Research Institute (GRI) grant funded by GIST, Korea in 2023.

Author information

Author notes

  1. Seungho Choi, Seong Hyeon Hong and Gwidong Han contributed equally to this work.

Authors and Affiliations

  1. School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, 61005, Korea

    Seungho Choi, Seong Hyeon Hong, Gwidong Han & Chunghee Cho

  2. R&D Center, KYNOGEN Corp, Suwon, Gyeonggi-do, Korea

    Seungho Choi

  3. RNA Therapeutics Institute, University of Massachusetts Chan Medical School, Worcester, MA, USA

    Seong Hyeon Hong

Authors
  1. Seungho Choi
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  2. Seong Hyeon Hong
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  3. Gwidong Han
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  4. Chunghee Cho
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Contributions

C.C. conceptualized the study. S.C. S.H.H. and G.H. produced, analyzed and curated the data. S.C., S.H.H. and C.C. wrote the draft. All authors have read and agreed to the final version of the manuscript. S.C., S.H.H. and G.H. contributed equally to this work.

Corresponding author

Correspondence to Chunghee Cho.

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Conflict of interest

The authors declare no conflict of interest.

Ethical approval

All animal experiments were performed in accordance with the Korean Food and Drug Administration (KFDA) guidelines. Protocols were reviewed and approved by the Institutional Animal Care and Use Committees (IACUC) of Gwangju Institute of Science and Technology (GIST) (permit number: GIST-2017-013).

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Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 Supplementary Figure 1: Distribution of expression values in the microarray data (PDF 141 KB)

13258_2023_1488_MOESM2_ESM.pdf

Supplementary file2 Supplementary Figure 2: Tissue distribution of human ortholgs of five testis-specific mouse genes (PDF 49 KB)

13258_2023_1488_MOESM3_ESM.xlsx

Supplementary file3 Supplementary Table 1: List of mRNAs expressed in testis, GC-1 cells, and GC-2 cells (XLSX 39573 KB)

Supplementary file4 Supplementary Table 2: List of mRNAs with testis-specificity information (XLSX 7473 KB)

Supplementary file5 Supplementary Table 3: Primers used for RT-PCR (PDF 270 KB)

13258_2023_1488_MOESM6_ESM.pdf

Supplementary file6 Supplementary Table 4: List of human orthologs and testis specificity of 25 selected testis-preferred mouse genes (PDF 187 KB)

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Choi, S., Hong, S.H., Han, G. et al. Profiling of testis-specific or testis-predominant genes expressed in mouse male germ cell lines GC-1 and GC-2. Genes Genom 46, 279–287 (2024). https://doi.org/10.1007/s13258-023-01488-1

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  • DOI: https://doi.org/10.1007/s13258-023-01488-1

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