Abstract:
Objective 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.
Methods 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.
Results 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.
Conclusions 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.
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