Citation: | JI Yunhua, WANG Linmeng, YAO Zhen, TAN Xiao, GUO Xuyan, HOU Haozhong, ZHANG Bo. Single-cell Transcriptome Analysis of Sertoli Cells under High-altitude Environment Reveals Reproductive Toxicity Mechanisms[J]. Medical Journal of Peking Union Medical College Hospital, 2024, 15(3): 587-597. DOI: 10.12290/xhyxzz.2023-0435 |
To construct a single-cell transcriptomic map of testicular tissue under hypobaric hypoxia exposure and perform diversity analysis of supportive cells, aiming to provide new insights into the mechanisms of reproductive toxicity for future research.
Twenty healthy male mice were randomly divided into a control group (n=10) and a hypobaric hypoxia group (n=10). The control group was raised under normal conditions, while the hypobaric hypoxia group was exposed to a low-pressure hypoxic environment(pressure=14 kPa, oxygen content=14.5%). After 6 weeks, testicular tissue from both groups of mice was collected, and the transcriptomic map was obtained using the Singleron MatrixTM single-cell platform and Illumina NovaSeq second-generation sequencing technology. Clustering, pseudo-temporal trajectory analysis, functional analysis, transcription factor and cellular communication research were conducted.
A single-cell transcriptomic map of testicular tissue was successfully constructed, including 6 samples with a total of approximately 49 027 cells covering 11 cell types. Supportive cells were clustered into 4 subgroups using non-negative matrix factorization algorithm. Subgroup 3 may be more sensitive to hypoxic stimulation and could reduce reproductive cell function damage caused by hypobaric hypoxia by affecting PTN expression levels and regulating the cell cycle of spermatogonia. The PTN-PTPR pathway may be an important regulatory node for supportive cells to regulate spermatogonia. Subgroup 4 could regulate the cell cycle of spermatogonia after exposure to hypobaric hypoxia through the KITLG-KIT signaling pathway, thus affecting sperm development.
Based on single-cell sequencing technology, the molecular basis and regulatory signals of supportive cells under hypobaric hypoxia exposure is revealed for the first time. It provides an in-depth exploration of the mechanisms of reproductive toxicity induced by hypobaric hypoxia at the single-cell level, offering a new perspective for future clinical research in this field.
[1] |
Bosco G, Paoli A, Rizzato A, et al. Body composition and endocrine adaptations to high-altitude trekking in the Himalayas[J]. Adv Exp Med Biol, 2019, 1211: 61-68.
|
[2] |
Alsup C, Lipman G S, Pomeranz D, et al. Interstitial pulmonary edema assessed by lung ultrasound on ascent to high altitude and slight association with acute mountain sickness: a prospective observational study[J]. High Alt Med Biol, 2019, 20(2): 150-156. DOI: 10.1089/ham.2018.0123
|
[3] |
Hofmann M C, Mcbeath E. Sertoli cell-germ cell interactions within the niche: paracrine and juxtacrine molecular communications[J]. Front Endocrinol (Lausanne), 2022, 13: 897062. DOI: 10.3389/fendo.2022.897062
|
[4] |
Chang D H, Kong F Y, Jiang W, et al. Effects of L-carni-tine administration on sperm and sex hormone levels in a male Wistar rat reproductive system injury model in a high-altitude hypobaric hypoxic environment[J]. Reprod Sci, 2023, 30(7): 2231-2247. DOI: 10.1007/s43032-022-00948-5
|
[5] |
Meroni S B, Galardo M N, Rindone G, et al. Molecular mechanisms and signaling pathways involved in Sertoli cell proliferation[J]. Front Endocrinol (Lausanne), 2019, 10: 224. DOI: 10.3389/fendo.2019.00224
|
[6] |
Peng Y J, Tang X T, Shu H S, et al. Sertoli cells are the source of stem cell factor for spermatogenesis[J]. Development, 2023, 150(6): dev200706. DOI: 10.1242/dev.200706
|
[7] |
Zhao Q Q, Sun X C, Zheng C, et al. The evolutionarily conserved hif-1/bnip3 pathway promotes mitophagy and mitochondrial fission in crustacean testes during hypoxia[J]. Am J Physiol Regul Integr Comp Physiol, 2023, 324(1): R128-R142. DOI: 10.1152/ajpregu.00212.2022
|
[8] |
Li X Y, Zhang M H, Chen Z W, et al. Male reproductive system and simulated high-altitude environment: preliminary results in rats[J]. Asian J Androl, 2023, 25(3): 426-432. DOI: 10.4103/aja202290
|
[9] |
He J, Cui J H, Wang R, et al. Exposure to hypoxia at high altitude (5380 m) for 1 year induces reversible effects on semen quality and serum reproductive hormone levels in young male adults[J]. High Alt Med Biol, 2015, 16(3): 216-222. DOI: 10.1089/ham.2014.1046
|
[10] |
Oyedokun P A, Akhigbe R E, Ajayi L O, et al. Impact of hypoxia on male reproductive functions[J]. Mol Cell Biochem, 2023, 478(4): 875-885. DOI: 10.1007/s11010-022-04559-1
|
[11] |
Zhang D C, Chen R, Cai Y H, et al. Hyperactive reactive oxygen species impair function of porcine Sertoli cells via suppression of surface protein ITGB1 and connexin-43[J]. Zool Res, 2020, 41(2): 203-207. DOI: 10.24272/j.issn.2095-8137.2020.024
|
[12] |
Van de Sande B, Flerin C, Davie K, et al. A scalable SCENIC workflow for single-cell gene regulatory network analysis[J]. Nat Protoc, 2020, 15(7): 2247-2276. DOI: 10.1038/s41596-020-0336-2
|
[13] |
Medina D L, Di Paola S, Peluso I, et al. Lysosomal calcium signalling regulates autophagy through calcineurin and TFEB[J]. Nat Cell Biol, 2015, 17(3): 288-299. DOI: 10.1038/ncb3114
|
[14] |
Shkreta L, Delannoy A, Salvetti A, et al. SRSF10: an atypical splicing regulator with critical roles in stress response, organ development, and viral replication[J]. RNA, 2021, 27(11): 1302-1317. DOI: 10.1261/rna.078879.121
|
[15] |
Liu W B, Lu X K, Zhao Z H, et al. SRSF10 is essential for progenitor spermatogonia expansion by regulating alternative splicing[J]. Elife, 2022, 11: e78211. DOI: 10.7554/eLife.78211
|
[16] |
Gerber J, Heinrich J, Brehm R. Blood-testis barrier and Sertoli cell function: lessons from SCCx43KO mice[J]. Reproduction, 2016, 151(2): R15-R27. DOI: 10.1530/REP-15-0366
|
[17] |
Meng X, Lindahl M, Hyvönen M E, et al. Regulation of cell fate decision of undifferentiated spermatogonia by GDNF[J]. Science, 2000, 287(5457): 1489-1493. DOI: 10.1126/science.287.5457.1489
|
[18] |
O'Donnell L, Smith L B, Rebourcet D. Sertoli cells as key drivers of testis function[J]. Semin Cell Dev Biol, 2022, 121: 2-9. DOI: 10.1016/j.semcdb.2021.06.016
|
[19] |
Yan R G, Yang Q L, Yang Q E. E4 transcription factor 1 (E4F1) regulates Sertoli cell proliferation and fertility in mice[J]. Animals (Basel), 2020, 10(9): 1691.
|
[20] |
Wang Y, Qiu B, Liu J, et al. Cocaine- and amphetamine-regulated transcript facilitates the neurite outgrowth in cortical neurons after oxygen and glucose deprivation through PTN-dependent pathway[J]. Neuroscience, 2014, 277: 103-110. DOI: 10.1016/j.neuroscience.2014.06.064
|
[21] |
Dong Z, Li C Y, Coates D. PTN-PTPRZ signalling is involved in deer antler stem cell regulation during tissue regeneration[J]. J Cell Physiol, 2021, 236(5): 3752-3769. DOI: 10.1002/jcp.30115
|
[22] |
Huang J C, Chen S C, Chang W A, et al. KITLG promotes glomerular endothelial cell injury in diabetic nephropathy by an autocrine effect[J]. Int J Mol Sci, 2022, 23(19): 11723. DOI: 10.3390/ijms231911723
|
[23] |
Cheng P, Chen H, Liu S R, et al. SNPs in KIT and KITLG genes may be associated with oligospermia in Chinese population[J]. Biomarkers, 2013, 18(8): 650-654. DOI: 10.3109/1354750X.2013.838307
|
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