Transcriptional and Translational Heterogeneity among Neonatal Mouse Spermatogonia1
Year: 2015
Type: article
Abstract: Spermatogonial stem cells (SSCs) are a subset of undifferentiated spermatogonia responsible for ongoing spermatogenesis in mammalian testes. Spermatogonial stem cells arise from morphologically homogeneous prospermatogonia, but growing evidence suggests that only a subset of prospermatogonia develops into the foundational SSC pool. This predicts that subtypes of undifferentiated spermatogonia with discrete mRNA and protein signatures should be distinguishable in neonatal testes. We used single-cell quantitative RT-PCR to examine mRNA levels of 172 genes in individual spermatogonia from 6-day postnatal (P6) mouse testes. Cells enriched from P6 testes using the StaPut or THY1(+) magnetic cell sorting methods exhibited considerable heterogeneity in the abundance of specific germ cell and stem cell mRNAs, segregating into one somatic and three distinct spermatogonial clusters. However, P6 Id4-eGFP(+) transgenic spermatogonia, which are known to be enriched for SSCs, were more homogeneous in their mRNA levels, exhibiting uniform levels for the majority of genes examined (122 of 172). Interestingly, these cells displayed nonuniform (50 of 172) expression of a smaller cohort of these genes, suggesting there is substantial heterogeneity even within the Id4-eGFP(+) population. Further, although immunofluorescence staining largely demonstrated conformity between mRNA and protein levels, some proteins were observed in patterns that were disparate from those detected for the corresponding mRNAs in Id4-eGFP(+) spermatogonia (e.g., Kit, Sohlh2, Stra8), suggesting additional heterogeneity is introduced at the posttranscriptional level. Taken together, these data demonstrate the existence of multiple spermatogonial subtypes in P6 mouse testes and raise the intriguing possibility that these subpopulations may correlate with the development of functionally distinct spermatogenic cell types.
Source: Biology of Reproduction
Language: English
Subfield: Reproductive Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: green
Funders: National Science Foundation, Max and Minnie Tomerlin Voelcker Fund, University of Texas at San Antonio, Robert J. Kleberg, Jr. and Helen C. Kleberg Foundation, National Institutes of Health, Langley Research Center, University of Texas Health Science Center at San Antonio, National Cancer Institute, National Institute of General Medical Sciences, National Center for Research Resources, National Center for Advancing Translational Sciences, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: G1060574484, G1114781936, Mays Cancer Center at UT Health SA, G156786057, Control of Spermatogonial Stem Cell Fate Decisions by HLH Factors, G3288338257, Institute for Integration of Medicine & Science: A Partnership to Improve Health, G4244661137, MRI: Acquisition of a BioMark HD System for single-cell genomics research., Regulation of spermatogonial stem cell proliferation and differentiation by UTF1, G6111067761, G6902769102, G7796852985, Transcriptional regulatory networks in spermatogonial stem cells, Analysis of a translation repression program during neonatal male germ cell development, Transcriptional regulatory networks in spermatogonial stem cells, G9000150992
OpenAlex ID: W2027706672