Transcriptome Profiles of Penaeus (Marsupenaeus) japonicus Animal and Vegetal Half-Embryos: Identification of Sex Determination, Germ Line, Mesoderm, and Other Developmental Genes

Melony J. Sellars, Carolyn Trewin, Sean M. McWilliam, R. S.E. Glaves, Philip L. Hertzler

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

There is virtually no knowledge of the molecular events controlling early embryogenesis in Penaeid shrimp. A combination of controlled spawning environment, shrimp embryo micro-dissection techniques, and next-generation sequencing was used to produce transcriptome EST datasets of Penaeus japonicus animal and vegetal half-embryos. Embryos were collected immediately after spawning, and then blastomeres were separated at the two-cell stage and allowed to develop to late gastrulation, then pooled for RNA isolation and cDNA synthesis. Ion Torrent sequencing of cDNA from approximately 500 pooled animal and vegetal half-embryos from multiple spawnings resulted in 560,516 and 493,703 reads, respectively. Reads from each library were assembled and Gene Ontogeny analysis produced 3479 annotated animal contigs and 4173 annotated vegetal contigs, with 159/139 hits for developmental processes in the animal/vegetal contigs, respectively. Contigs were subject to BLAST for selected developmental toolbox genes. Some of the genes found included the sex determination genes sex-lethal and transformer; the germ line genes argonaute 1, boule, germ cell-less, gustavus, maelstrom, mex-3, par-1, pumilio, SmB, staufen, and tudor; the mesoderm genes brachyury, mef2, snail, and twist; the axis determination/segmentation genes β-catenin, deformed, distal-less, engrailed, giant, hairy, hunchback, kruppel, orthodenticle, patched, tailless, and wingless/wnt-8c; and a number of cell-cycle regulators. Animal and vegetal contigs were computationally subtracted from each other to produce sets unique to either half-embryo library. Genes expressed only in the animal half included bmp1, kruppel, maelstrom, and orthodenticle. Genes expressed only in the vegetal half included boule, brachyury, deformed, dorsal, engrailed, hunchback, spalt, twist, and wingless/wnt-8c.

Original languageEnglish
Pages (from-to)252-265
Number of pages14
JournalMarine Biotechnology
Volume17
Issue number3
DOIs
StatePublished - Jun 1 2015

Keywords

  • Embryo transcriptome
  • Germ line
  • Kuruma shrimp
  • Mesoderm
  • Penaeus japonicus
  • Sex determination

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