A multistate stem cell dynamics maintains homeostasis in mouse spermatogenesis

Abstract

Highlights d Single pool of heterogenous stem cells supports homeostasis in mouse spermatogenesis d Stem cells move reversibly between renewal-biased and differentiation-primed states d Stem cell dynamics depends on distinct rates of state transition and cell division d Such multistate dynamics reduces mitotic load, while keeping stem cell density high

A scRNA-seq Approach to Identifying Changes in Spermatogonial Stem Cell Gene Expression Following in vitro Culture

“…It was noted that progenitor spermatogonia fell in to two 'sub-categories' in which they either displayed dual expression of Sox3 and Neurog3 (denoted 'Progenitor A', capturing clusters 5, 8 and 9), or expressed Sox3 only, along with increased expression of Stra8 (denoted 'Progenitor B', capturing clusters 3, 6, 7 and 10) (Figure 1E; Supplementary Figure S2B). This categorisation aligns with SOX3+/NEUROG3+ and SOX3+/NEUROG3subsets of progenitors that have been previously identified in the testes using antibody-based techniques (Nakagawa et al, 2021).…”

“…As a whole, the expression of a broad range of SSC markers (Etv5, Gfra1, Lhx1, Bcl6b, Ret) was robust across all SSC clusters (Figure 2C), noting that expression of Shisa6 (Tokue et al, 2017) was below levels of detection (Figure 2C), and Id4 levels could not reliably be assessed given that spermatogonia from the testis, but not from culture, came from an Id4-eGfp transgenic mouse line (Helsel et al, 2017b), with transgene expression causing an artefactual increase in expression in the testis dataset (Martin and Wang, 2011). It is interesting to note, however, that cluster SSC_0 exhibited a small but significant (p < 0.001) increase in expression of a subset of genes that have been reported to play a role in SSC maintenance, including; Etv5 (Wu et al, 2011), Gfra1 (He et al, 2007, Ret (He et al, 2007), Sall4 (Chan et al, 2017), Chd4 (Cafe et al, 2021b, and Plvap (Nakagawa et al, 2021) (Figure 2C; Supplementary Dataset S1). In considering that 49 spermatogonia from culture (representing 6.8% of total cells from the culture dataset) reside within the SSC_0 cluster, one could postulate that these may be the SSCs that colonise recipient testes post-transplantation.…”

Identification of quiescent FOXC2+spermatogonial stem cells in adult mammals

“…In recent years, great insights into SSCs behaviors and regulations have been provided by a body of pioneer works, especially with recent advances in single-cell gene-expression profiling, highlighting great heterogeneity of SSCs and focusing on characterizing the nature of SSCs states especially for seeking the primitive subgroup among them. Within the population of uSPG, a number of genes relatively higher expressed in primitive subfractions have been identified and well investigated, i.e., Gfra1, ID4, Ret, Eomes, Pax7, Nanos2, Shisa6, T, Pdx1, Lhx1, Egr2 and Plvap (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15). Particularly, Gfra1, ID4, Eomes, Pax7, Nanos2, and Plvap are further validated as the SSCs markers through lineage tracing experiment, which is considered to be a reliable method to study the origin and development of stem cells.…”

Quantitative proteomic analysis based on tandem mass tagging (TMT) reveals key proteins related to DNA hydroxymethylation enzyme TET1 for spermatogonia self-renewal

“…PLZF is a transcriptional repressor that regulates the epigenetic state of undifferentiated cells and is the rst gene in germ cells that has been shown to be essential for self-renewal of mammalian stem cells [22,23]. GFRα1 is a growth factor required for SSC self-renewal and survival [24,25]. Proliferating cell nuclear antigen (PCNA) is known as a molecular marker of proliferation because of its role in replication [26,27].…”