comparison-sperm-dna-analysis-outcomes-using-different-gating

Comparison of Sperm DNA Analysis Outcomes Using Different Gating Strategies in Flow Cytometry

Sperm DNA integrity is a key factor in ensuring successful fertilization, embryo development, and ongoing&#160;pregnancy. Sperm chromatin structure ...

The Affiliated Huai'an No. 1 People's Hospital of Nanjing Medical University

Thousands of genes are expressed in the testis of mice. However, the details about their roles during spermatogenesis have not been well-clarified for most genes. The purpose of this study was to examine the effect of  deficiency on mouse spermatogenesis and male fertility. -knockout (KO) mice were generated using CRISPR/Cas9 technology on C57BL/6J background. We found no obvious differences between -KO and -WT mice in fertility tests, testicular weight, sperm concentrations, or morphology. Histological analysis found that -KO mouse testes had normal germ cell types and mature sperm. These findings indicated that  was dispensable for male fertility in mice. Our results may save time and resources by allowing other researchers to focus on genes that are more meaningful for fertility studies. We also found that mRNAs of two  family members ( and ) were expressed on higher mean levels in -KO total mouse testes, compared to -WT mice. This effect was not found in mouse GC-1 and GC-2 germ cell lines with the  gene transiently knocked down. This result may indicate that a gene compensation phenomenon was present in the testes of -KO mice.

 is not essential for spermatogenesis and male fertility in mice.

The testis supports spermatogenesis through a tightly regulated microenvironment, and the bacterial microbiome (BM) may influence host cells through immune and metabolic pathways, thereby impacting reproductive capacity. Here, we applied invasion-adhesion-directed expression sequencing (INVADE-seq), a single-cell RNA sequencing approach that simultaneously captures host and bacterial transcripts, to examine how bacterial signals shape testicular cell states. We detected a sparse but widespread bacterial presence across multiple cell types, with somatic and early germ cells outside the blood-testis barrier (BTB) showing relatively higher bacterial abundance. Bacterial load increased with age, coinciding with transcriptional signatures of reduced BTB function. At the cellular level, bacterial-positive Leydig cells exhibited activation of steroidogenic genes, whereas macrophages upregulated pathways related to autophagy and immune modulation. These findings not only deepen our understanding of testicular microbiome biology but also hold promise for the discovery of novel diagnostic biomarkers and therapeutic targets for BM-related and age-associated male subfertility.

Article	Year
is not essential for spermatogenesis and male fertility in mice. PeerJ\| PubMed ID: 38111663	2023
Single-cell analysis of testicular bacterial microbiome changes during aging and effect on reproductive capacity in mice. iScience\| PubMed ID: 41438041	2025

Yu Qiao

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