High-throughput analysis reveals novel maternal germline rnas crucial for primordial germ cell preservation and proper migration

Dawn A. Owens, Amanda M. Butler, Tristan H. Aguero, Karen M. Newman, Derek Van Booven, Mary Lou King

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

During oogenesis, hundreds of maternal RNAs are selectively localized to the animal or vegetal pole, including determinants of somatic and germline fates. Although microarray analysis has identified localized determinants, it is not comprehensive and is limited to known transcripts. Here, we utilized high-throughput RNAsequencing analysis to comprehensively interrogate animal and vegetal pole RNAs in the fully grown Xenopus laevis oocyte. We identified 411 (198 annotated) and 27 (15 annotated) enriched mRNAs at the vegetal and animal pole, respectively. Ninety were novel mRNAs over 4-fold enriched at the vegetal pole and six were over 10-fold enriched at the animal pole. Unlike mRNAs, microRNAs were not asymmetrically distributed. Whole-mount in situ hybridization confirmed that all 17 selected mRNAs were localized. Biological function and network analysis of vegetally enriched transcripts identified protein-modifying enzymes, receptors, ligands, RNA-binding proteins, transcription factors and co-factors with five defining hubs linking 47 genes in a network. Initial functional studies of maternal vegetally localized mRNAs show that sox7 plays a novel and important role in primordial germ cell (PGC) development and that ephrinB1 (efnb1) is required for proper PGC migration. We propose potential pathways operating at the vegetal pole that highlight where future investigations might be most fruitful.

Original languageEnglish (US)
Pages (from-to)292-304
Number of pages13
JournalDevelopment (Cambridge)
Volume144
Issue number2
DOIs
StatePublished - Jan 15 2017

Keywords

  • Gene pathways
  • Localized RNAs
  • PGCs
  • RNA-seq
  • Scaffold
  • Xenopus

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology

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