Identification of lineage-specific zygotic transcripts in early Caenorhabditis elegans embryos

Scott M. Robertson, Premnath Shetty, Rueyling Lin

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

During Caenorhabditis elegans embryogenesis, a maternally supplied transcription factor, SKN-1, is required for the specification of the mesendodermal precursor, EMS, in the 4-cell stage embryo. When EMS divides, it gives rise to a mesoderm-restricted precursor, MS, and an endoderm-restricted precursor, E. To systematically identify genes that function as key regulators of MS and/or E-derived tissues, we identified, by microarray analyses, genes that are newly transcribed within a short developmental window (approximately 30 min) encompassing the generation and fate specification of the MS and E blastomeres. By comparing total cDNAs generated from individual, carefully staged embryos, we identified 275 genes up-regulated in 12-cell embryos compared to 4-cell embryos. Fifty of these 275 genes are down-regulated in 12-cell skn-1 mutant embryos and are designated skn-1-dependent zygotic (sdz) genes. The spatial and temporal expression patterns in C. elegans embryos of 10 randomly selected sdz genes were analyzed by a nuclear GFP reporter driven by the endogenous 5′ regulatory sequence of each gene. GFP expression, although absent at the 4-cell stage, was detected at the 12- to 16-cell stage for all 10 genes and was restricted to EMS-derived lineages for 7 of the 10. Among the seven lineage-specific genes, three genes are expressed equally in both MS and E lineages, two are expressed exclusively or predominantly in the MS lineage, and two are expressed exclusively in the E lineage. Depletion of skn-1 by RNAi abolishes the expression of all seven reporter transgenes in vivo, confirming that these genes are indeed skn-1 dependent. These results demonstrate the successful combination of single-staged embryo cDNAs, genetic mutants, and whole transcriptome microarray analysis to identify stage- and lineage-specific transcripts in early C. elegans embryos.

Original languageEnglish (US)
Pages (from-to)493-507
Number of pages15
JournalDevelopmental Biology
Volume276
Issue number2
DOIs
StatePublished - Dec 15 2004

Fingerprint

Caenorhabditis elegans
Embryonic Structures
Genes
Complementary DNA
Tissue Array Analysis
Blastomeres
Endoderm
Gene Expression Profiling
Mesoderm
Microarray Analysis
RNA Interference
Transgenes
Embryonic Development
Transcription Factors

Keywords

  • C. elegans
  • Embryos
  • Lineage specific
  • Microarray
  • skn-1
  • Zygotic

ASJC Scopus subject areas

  • Developmental Biology

Cite this

Identification of lineage-specific zygotic transcripts in early Caenorhabditis elegans embryos. / Robertson, Scott M.; Shetty, Premnath; Lin, Rueyling.

In: Developmental Biology, Vol. 276, No. 2, 15.12.2004, p. 493-507.

Research output: Contribution to journalArticle

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