Journal Article

Comprehensive profiling of migratory primordial germ cells reveals niche-specific differences in non-canonical Wnt and Nodal-Lefty signaling in anterior vs posterior migrants

Zussman, Jay W, Wagner, Daniel E, Laird, Diana J, Jaszczak, Rebecca Garrett

2025

Overview

Mammalian primordial germ cells (PGCs) migrate asynchronously through the embryonic hindgut and dorsal mesentery to reach the gonads. We previously found that interaction with different somatic niches regulates mouse PGC proliferation along the migration route. To characterize transcriptional heterogeneity of migrating PGCs and their niches, we performed single-cell RNA sequencing of 13,262 mouse PGCs and 7868 surrounding somatic cells during migration (E9.5, E10.5, E11.5) and in anterior vs posterior locations to enrich for leading and lagging migrants. Analysis of PGCs by position revealed dynamic gene expression changes between faster or earlier migrants in the anterior and slower or later migrants in the posterior at E9.5; these differences include migration-associated actin polymerization machinery and epigenetic reprogramming-associated genes. We furthermore identified changes in signaling with various somatic niches, notably strengthened interactions with hindgut epithelium via non-canonical WNT (ncWNT) in posterior PGCs compared to anterior. Reanalysis of a previously published dataset suggests that ncWNT signaling from the hindgut epithelium to early migratory PGCs is conserved in humans. Trajectory inference methods identified putative differentiation trajectories linking cell states across timepoints and from posterior to anterior in our mouse dataset. At E9.5, we mainly observed differences in cell adhesion and actin cytoskeletal dynamics between E9.5 posterior and anterior migrants. At E10.5, we observed divergent gene expression patterns between putative differentiation trajectories from posterior to anterior, including Nodal signaling response genes Lefty1, Lefty2, and Pycr2 and reprogramming factors Dnmt1, Prc1, and Tet1 . At E10.5, we experimentally validated anterior migrant-specific Lefty1/2 upregulation via whole-mount immunofluorescence staining for LEFTY1/2 and phosphorylated SMAD2/3, suggesting that elevated autocrine Nodal signaling in migrating PGCs occurs as they near the gonadal ridges. Together, this positional and temporal atlas of mouse PGCs supports the idea that niche interactions along the migratory route elicit changes in proliferation, actin dynamics, pluripotency, and epigenetic reprogramming.

• Publisher: eLife Science Publications, Ltd,eLife Sciences Publications Ltd,eLife Sciences Publications, Ltd
• Subject: Actin / Animals / Approximation / Autocrine signalling / Behavior / Cell adhesion / Cell differentiation / cell heterogeneity / Cell migration / Cell Movement / Cells / Cytoskeleton / Datasets / Developmental Biology / DNMT1 protein / Embryos / Epigenetic inheritance / Epigenetics / Epithelium / Gene expression / Gene Expression Regulation, Developmental / Genes / Genetic transcription / Germ cells / Germ Cells - metabolism / Germ Cells - physiology / Gonads / Hindgut / Immunofluorescence / Left-Right Determination Factors - genetics / Left-Right Determination Factors - metabolism / Mesentery / Methyltransferases / Mice / Migration / Nodal / Nodal Protein - genetics / Nodal Protein - metabolism / Ontology / Pluripotency / primordial germ cell / RNA sequencing / Signal Transduction / Smad2 protein / Somatic cells / Wnt / Wnt protein / Wnt Signaling Pathway