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. 2022 Dec 19;11(24):4135.
doi: 10.3390/cells11244135.

Single-Cell RNAseq Resolve the Potential Effects of LanCL1 Gene in the Mouse Testis

Jiangting Lu  1 , Jinling Liao  2 , Min Qin  3 , Hui Li  1 , Qingyuan Zhang  1 , Yang Chen  1   2 , Jiwen Cheng  1   2
Affiliations

Single-Cell RNAseq Resolve the Potential Effects of LanCL1 Gene in the Mouse Testis

Jiangting Lu et al. Cells. .

Abstract

Infertility affects lots of couples, half of which are caused by male factors. The LanCL1 gene is highly expressed in testis specifically, which might affect the development of sperms. In order to understand the potential functions of the LanCL1 gene in the testis, this study was conducted with constructed transgenic LanCL1 knockout mice. The mouse breeding experiment, semen analysis and single-cell RNAseq of testicular tissue were performed. Results suggested that the LanCL1 gene would significantly influence the reproduction ability and sperm motility of male mice. Single-cell RNAseq also confirmed the high expression of the LanCL1 gene in the spermatocytes and spermatids. Downregulating the LanCL1 gene expression could promote M2 macrophage polarity to maintain testicular homeostasis. Moreover, the LanCL1 gene could affect both the germ cells and stromal cells through various pathways such as the P53 signaling and the PPAR signaling pathway to disturb the normal process of spermatogenesis. However, no effects of the LanCL1 gene in testosterone synthesis and serum testosterone level were shown. Further studies are needed to discuss the mechanisms of the LanCL1 gene in the various cells of the testis independently.

Keywords: LanCL1; male infertility; single-cell RNAseq; sperm; transgenic mice.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
LanCL1 gene knockout influences the semen quality. (a) The expression of mRNA and protein in the LanCL1+/+, LanCL1+/− and LanCL1−/− mice. (b) Parameters of semen quality in the LanCL1+/+ and LanCL1−/− mice. (c) Sperm morphology.
Figure 2
Figure 2
Single-cell RNAseq of the testis identified the high expression of the Cir1 gene in LanCL1−/− mice. (a) UAMP plot of the cell clusters. (b) Heatmap of the marker genes in the cell clusters. (c) The LanCL1 gene expression in the cell clusters. (d) Immunofluorescence of the LanCL1 gene in the testicular tissue. (e) The volcano plot of the DEGs in the LanCL1+/+ and LanCL1−/− mice. (f) Venn diagram of up-regulation in the Germ cells of the LanCL1−/− cell.
Figure 3
Figure 3
The LanCL1 gene maintains the development of normal sperm motility in germ cells. (a) The percent of cell numbers in every cell cluster from LanCL1+/+ and LanCL1−/− mice. (b) UAMP plot of the cluster 7. (c) The cell percent of new clusters. (d) GO annotation of the DEGs in the LanCL1−/− and LanCL1+/+ mice.
Figure 4
Figure 4
The cell interactions signaling in various cell clusters regulated by the LanCL1 gene. (a) The cell communication before and after LanCL1 gene knockout. (b) The alteration of signaling pathway. (c) Heatmap of the active signaling pathway in every cell clusters. (d) PDGF signaling pathway. (e) GALECTIN signaling pathway. (f) The M1 and M2 macrophage marker genes.
Figure 5
Figure 5
LanCL1 gene regulated the P53 and PPAR signaling other than the testosterone synthesis in the Leyding cells. (a) The numbers of DEGs in every cell clusters. (b) The GSEA of the DEGs in the LanCL1+/+ and LanCL1−/− mice of Leydig cell. (c) The steroid biosynthesis. (d) The marker genes of testosterone biosynthesis. (e) Serum testosterone level in the LanCL1+/+ and LanCL1−/− mice.
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References

    1. Duvuru R., Halabi M., Omolaoye T.S., Du Plessis S.S. The genetic causes of male infertility: A Middle East and North Africa perspective. F1000Research. 2022;11:125. doi: 10.12688/f1000research.106950.2. - DOI - PMC - PubMed
    1. Vander Borght M., Wyns C. Fertility and infertility: Definition and epidemiology. Clin. Biochem. 2018;62:2–10. doi: 10.1016/j.clinbiochem.2018.03.012. - DOI - PubMed
    1. Agarwal A., Leisegang K., Majzoub A., Henkel R., Finelli R., Panner Selvam M.K., Tadros N., Parekh N., Ko E.Y., Cho C.L., et al. Utility of Antioxidants in the Treatment of Male Infertility: Clinical Guidelines Based on a Systematic Review and Analysis of Evidence. World J. Men’s Health. 2021;39:233–290. doi: 10.5534/wjmh.200196. - DOI - PMC - PubMed
    1. Carlsen E., Giwercman A., Keiding N., Skakkebaek N.E. Evidence for decreasing quality of semen during past 50 years. BMJ. 1992;305:609–613. doi: 10.1136/bmj.305.6854.609. - DOI - PMC - PubMed
    1. Levine H., Jørgensen N., Martino-Andrade A., Mendiola J., Weksler-Derri D., Jolles M., Pinotti R., Swan S.H. Temporal trends in sperm count: A systematic review and meta-regression analysis of samples collected globally in the 20th and 21st centuries. Hum. Reprod. Update. 2022:dmac035. doi: 10.1093/humupd/dmac035. - DOI - PubMed

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