Haploid Medaka Stem Cells: Although diploid embryonic stem cells have been generated by various means, there would also be value in deriving haploid stem cells. In these cells, recessive mutations in essential genes would show phenotypes that would not be apparent in heterozygous animals. Yi et al. (p. 430 ) used the medaka fish model system to generate haploid stem cells that show stable growth and pluripotency. In addition, a fertile female medaka fish was produced by haploid embryonic stem cell nuclear transfer into a normal egg. This system has potential for analyzing recessive genes, for example, in disease phenotypes or in various cell lineages in culture.

Classic T cell subsets are defined by a small set of cell surface markers, while single-cell RNA sequencing (scRNA-seq) clusters cells using genome-wide gene expression profiles. The relationship between scRNA-seq clustered populations (scCPops) and cell surface marker–defined classic T cell subsets remains unclear. In this article, we integrated six bead-enriched T cell subsets with 62,235 single-cell transcriptomes from human PBMCs and clustered them into nine scCPops. Bead-enriched CD4⁺/CD45RA⁺/CD25⁻ naive T and CD8⁺/CD45RA⁺ naive T cells were mainly clustered into their scCPop counterparts, while cells from the other T cell subsets were assigned to multiple scCPops, including mucosal-associated invariant T cells and NKT cells. The multiple T cell subsets forming one scCPop exhibit similar expression patterns, but not vice versa, indicating scCPop is a more homogeneous cell population with similar cell states. Interestingly, we discovered and named IFN signaling–associated gene (ISAG) high T (ISAG^hi T) cells, a T cell subpopulation that highly expressed ISAGs. We further enriched ISAG^hi T cells from human PBMCs by FACS of BST2 for scRNA-seq analyses. The ISAG^hi T cell cluster disappeared on t-distributed stochastic neighbor embedding plot after removing ISAGs, whereas the ISAG^hi T cell cluster showed up by analysis of ISAGs alone, indicating ISAGs are the major contributor of the ISAG^hi T cell cluster. BST2⁺ and BST2⁻ T cells showing different efficiencies of T cell activation indicate that a high level of ISAGs may contribute to quick immune responses. Key Points Naive T cell showed the highest similarity to its scCPop counterpart in T cell subsets. Cell population identified by scRNA-seq is more homogeneous than classic T subset. ISAG^hi T were identified by scRNA-seq, which may contribute to quick immune response.

Primordial germ cell (PGC) specification occurs early in development. PGC specifiers have been identified in Drosophila, mouse, and human but remained elusive in most animals. Here we identify the RNA-binding protein Dnd as a critical PGC specifier in the medaka fish (Oryzias latipes). Dnd depletion specifically abolished PGCs, and its overexpression boosted PGCs. We established a single-cell culture procedure enabling lineage tracing in vitro. We show that individual blastomeres from cleavage embryos at the 32- and 64-cell stages are capable of PGC production in culture. Importantly, Dnd overexpression increases PGCs via increasing PGC precursors. Strikingly, dnd RNA forms prominent particles that segregate asymmetrically. Dnd concentrates in germ plasm and stabilizes germ plasm RNA. Therefore, Dnd is a critical specifier of fish PGCs and utilizes particle partition as a previously unidentified mechanism for asymmetric segregation. These findings offer insights into PGC specification and manipulation in medaka as a lower vertebrate model.