{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T08:36:52Z","timestamp":1790671012365,"version":"4.1.0"},"reference-count":34,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T00:00:00Z","timestamp":1768176000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Reprod. Health"],"abstract":"<jats:p>\n                    The molecular basis of spermatogenesis, which is a tightly regulated spatiotemporal process in testicular seminiferous tubules where germ cell differentiation and somatic-germ cell interactions drive sperm production, remains incompletely understood. Histological staining techniques lack molecular resolution, while scRNA-seq loses spatial context. Conventional spatial transcriptomics (approx. 100\u2005\u03bcm resolution) is too coarse-grained for testicular cells (10\u201320\u2005\u03bcm in diameter), leading to mixed-cell signals. In this study, we used Salus-STS high-resolution spatial transcriptomics (\u223c1\u2005\u03bcm resolution) and Salus Cellbins Algorithm to characterize the spatial transcriptomic profile of mouse testes at single-cell level. Integrating the spatial data with scRNA-seq via RCTD annotated major cell subtypes, whose distributions aligned with histology. Pseudotime and spatial gradient analyses revealed a basement membrane-to-lumen developmental axis, with luminal genes (e.g.,\n                    <jats:italic>Prm2<\/jats:italic>\n                    ) enriched in sperm maturation and basal genes (e.g.,\n                    <jats:italic>mt-Nd4<\/jats:italic>\n                    ) linked to mitochondrial metabolism\u2014validated by PPI and GO analyses. The biological relevance of these marker genes is underscored by the fact that mutations in\n                    <jats:italic>Prm2<\/jats:italic>\n                    and\n                    <jats:italic>mt-Nd4<\/jats:italic>\n                    are known to be associated with human male infertility, highlighting their potential diagnostic value. This work enables high-resolution dissection of spermatogenesis\u2019 spatiotemporal dynamics, providing insights into male reproductive biology.\n                  <\/jats:p>","DOI":"10.3389\/frph.2025.1747902","type":"journal-article","created":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T07:52:03Z","timestamp":1768204323000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Spatiotemporal dynamics of spermatogenesis: insights from high-resolution spatial transcriptomics and pseudotime trajectories in mouse 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