ArticleCytometry. Part A : the journal of the International Society for Analytical Cytology2014
Optimized flow cytometry isolation of murine spermatocytes.
Article in Cytometry. Part A : the journal of the International Society for Analytical Cytology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 82 papers.
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Who cites it
82 citing papers in PubMed.
- PSMA8-Containing 20S Proteasome Regulates Spermiogenesis and Male Fertility.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Deciphering RNA modification dynamics during spermatogenesis and sperm maturation.Science China. Life sciences · 2026Article
- Folate-dependent one-carbon metabolism controls meiotic and post-meiotic epigenome remodeling in the male germline.bioRxiv : the preprint server for biology · 2026Article
- Nucleolar migration regulates meiotic sex chromosome inactivation via phase separation during mammalian spermatogenesis.Nature communications · 2026Article
- DAXX governs the silencing of LINE1 during spermatogenesis in mice.Nucleic acids research · 2026Article
- EIF5A Couples Translational Control With Transcriptional Reprogramming Through Chromocenter Reorganization During Spermiogenesis.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Acute testicular hyperthermia leads to rapid loss of global piRNA and a concurrent increase in LINE1 activity within heat sensitive male germ cells.Mobile DNA · 2026Article
- Maternal CENP-C restores centromere symmetry in mammalian zygotes to ensure proper chromosome segregation.Developmental cell · 2026Article
- Development and validation of a deep learning model based on cascade mask regional convolutional neural network to noninvasively and accurately identify human round spermatids.Journal of advanced research · 2026Article
- Acquisition of ampliconic sequences marks a selfish mouse t-haplotype.Nature communications · 2025Article
- Spatiotemporal mapping reveals Ccl8Clinical and translational medicine · 2025Article
- CENP-A is diluted during bovine spermatogenesis and is maintained at internally positioned centromere clusters in mature bull sperm.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2025Article
- Compensation for X-linked Pdha1 silencing by Pdha2 is essential for meiotic double-strand break repair in spermatogenesis.Development (Cambridge, England) · 2025Article
- The histone H3 lysine 36 demethylase KDM2A/FBXL11 controls Polycomb-mediated gene repression and germ cell development in male mice.Nature communications · 2025Article
- Mechanisms and reversibility of nicotine-induced spermatogenesis impairment and DNA methylation changes.Communications biology · 2025Article
- UBL7 is indispensable for spermiogenesis through protecting critical factors from excessive degradation by proteasomes.Nature communications · 2025Article
- The nuclear exosome co-factor MTR4 shapes the transcriptome for meiotic initiation.Nature communications · 2025Article
- Pluripotency factor Tex10 finetunes Wnt signaling for spermatogenesis and primordial germ cell development.Nature communications · 2025Article
- DNA methylation modulates nucleosome retention in sperm and H3K4 methylation deposition in early mouse embryos.Nature communications · 2025Article
- Isolation of Germ Cell Populations from Mouse Testis by Flow Cytometry.Methods in molecular biology (Clifton, N.J.) · 2025Article
22 more citing papers are in PubMed but not listed here.
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Abstract
Meiotic prophase I (MPI), is an initial stage of meiosis characterized by intricate homologous chromosome interactions, synapsis, and DNA recombination. These processes depend on the complex, but poorly understood early MPI events of homologous chromosome search, alignment, and pairing. Detailed molecular investigation of these early events requires isolation of individual MPI substages. Enrichment for Pachytene (P) and Diplotene (D) substages of late MPI was previously accomplished using flow cytometry. However, separation of early MPI spermatocytes, specifically, of Leptotene (L) and Zygotene (Z) substages, has been a challenge due to these cells' similar characteristics. In this report, we describe an optimized Hoechst-33342 (Hoechst)-based flow cytometry approach for isolating individual MPI populations from adult mouse testis. We get significant enrichment for individual L and Z spermatocytes, previously inseparable from each other, and optimize the isolation of other MPI substages. Our flow cytometry approach is a combination of three optimized strategies. The first is optimization of testis dissociation protocol that yields more consistent and reproducible testicular single cell suspension. The second involves optimization of flow cytometric gating protocol where a critical addition to the standard protocol for cell discrimination based on Hoechst fluorescence, involves a back-gating technique based on light scattering parameters. This step specifies selection of individual MPI substages. The third, is an addition of DNA content restriction to the gating protocol to minimize contamination from non-meiotic cells. Finally, we confirm significant enrichment of high-purity Preleptotene (PreL), L, Z, P, and D MPI spermatocytes using stage-specific marker distribution. The technique will facilitate understanding of the molecular events underlying MPI.
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