Abstract

Primordial germ cells (PGCs) are undifferentiated gametes with heterogeneity, an evolutionarily conserved characteristic across various organisms. 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. In this study, we sought to evaluate PGC heterogeneity in zebra finch using a single-cell RNA sequencing (scRNA-seq) approach. 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. We found that PGCs previously considered a single population can be separated into three subtypes showing differences in apoptosis, proliferation, and other biological processes. 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. Additionally, we discovered a novel biomarker, SMC1B, for gonadal PGCs in zebra finch. The results provide the first evidence of substantial heterogeneity in PGCs previously considered a single population in birds. This discovery expands our understanding of PGCs to avian species, and provides a basis for further research.

Highlights

  • The heterogeneity of primordial germ cells (PGCs), the only cells capable of transferring genetic information to the generation, during early embryonic development is conserved across a wide range of taxa (Han, 2009; Nguyen et al, 2019)

  • In clusters c1 to c17, the proportion of cells derived from males was relatively high, whereas c15, c18, and c19 showed a relatively high proportion of cells derived from females (Supplementary Figures S2B, S2C)

  • We demonstrated that four novel markers, SMC1B, RNF17, RBM46, and HORMAD1, as well as representative markers commonly used to identify Primordial germ cells (PGCs) from gonads in avian species, such as DAZL, DDX4, DND1, and PIWIL1, can be applied to zebra finch (Figure 4A)

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Summary

Introduction

The heterogeneity of primordial germ cells (PGCs), the only cells capable of transferring genetic information to the generation, during early embryonic development is conserved across a wide range of taxa (Han, 2009; Nguyen et al, 2019). PGCs showing aberrant migration are eliminated by apoptosis (Koprunner et al, 2001; Weidinger et al, 2003; Reichman-Fried et al, 2004). 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). The heterogeneity of PGCs can be caused by differences in proliferation, as observed in flies (Drosophila melanogaster)

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