![]() Morphological analyses of germ cell heterogeneity and genetic manipulation of germ cells have been accelerated by development of single-cell RNA sequencing (scRNA-seq) techniques, which enable transcriptome studies and detailed lineage specification. PGCs with the potential to become germ cell tumors and harboring deleterious mutations can be eliminated, and only a subset of the initial population completes fetal development ( Runyan et al., 2008). Highly proliferative PGC populations are overrepresented in the germline and may compete for access to signaling factors essential for development ( Gilboa and Lehmann, 2006). ![]() The heterogeneity of PGCs can also be caused by differences in proliferation, as observed in flies ( Drosophila melanogaster). In mice ( Mus musculus), PGCs show asynchronous migration, in which some cells lag behind or move to an ectopic location via niche signals or intrinsic heterogeneity aberrant migration is closely related to cell survival ( Molyneaux et al., 2001 Laird et al., 2011 Cantú et al., 2016). PGCs showing aberrant migration are eliminated by apoptosis ( Koprunner et al., 2001 Weidinger et al., 2003 Reichman-Fried et al., 2004). In zebrafish ( Danio rerio), chemokine responsiveness and motility, which determine migration toward gonads during early development, differ among individual cells. Initial PGCs are heterogeneous with respect to their contribution to germline development ( Ueno et al., 2009). The heterogeneity of primordial germ cells (PGCs), the only cells capable of transferring genetic information to the next generation, during early embryonic development is conserved across a wide range of taxa ( Han, 2009 Nguyen et al., 2019). This discovery expands our understanding of PGCs to avian species, and provides a basis for further research. The results provide the first evidence of substantial heterogeneity in PGCs previously considered a single population in birds. Additionally, we discovered a novel biomarker, SMC1B, for gonadal PGCs in zebra finch. The three PGC subtypes were specifically enriched for genes showing expression patterns related to germness or pluripotency, suggesting functional differences in PGCs according to the three subtypes. We found that PGCs previously considered a single population can be separated into three subtypes showing differences in apoptosis, proliferation, and other biological processes. Using scRNA-seq of embryonic gonadal cells from male and female zebra finches at Hamburger and Hamilton (HH) stage 28, we annotated nine cell types from 20 cell clusters. In this study, we sought to evaluate PGC heterogeneity in zebra finch using a single-cell RNA sequencing (scRNA-seq) approach. ![]() Although dynamic selection at the level of early germ cell populations is an important biological feature linked to fertility, the heterogeneity of PGCs in avian species has not been characterized. Primordial germ cells (PGCs) are undifferentiated gametes with heterogeneity, an evolutionarily conserved characteristic across various organisms. 2Department of Computer Convergence Software, Korea University, Sejong, South Korea.1Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, Seoul, South Korea.Kyung Min Jung 1 †, Minseok Seo 2 †, Young Min Kim 1, Jin Lee Kim 1 and Jae Yong Han 1*
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