Dissecting mammalian reproduction with spatial transcriptomics.

This article reviews the application of spatial transcriptomics (ST) technologies—methods that preserve spatial context while profiling gene expression—to advance understanding of mammalian reproduction, including gametogenesis, embryogenesis, and reproductive pathologies. It summarizes targeted and unbiased ST approaches such as in situ hybridization (ISH), in situ sequencing (ISS), and solid-phase capture methods, highlighting their principles, advantages, limitations, and costs. The review details biological insights gained from ST studies, including spatial gene expression patterns in testicular and uterine tissues, cellular neighborhoods influencing reproductive physiology, and altered microenvironments in reproductive diseases like diabetes-induced testicular injury, prostate and ovarian cancers, and endometriosis. It also discusses experimental and computational challenges in applying ST to reproductive research and outlines future directions involving temporal dynamics, multi-omics integration, and spatially resolved gene perturbation analyses. This synthesis aims to inform reproductive biologists and clinicians about current ST technologies and their potential to enhance both basic and clinical reproductive research.