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

• Published in: Marine biotechnology (New York, N.Y.) Vol. 17; no. 3; pp. 252 - 265
• Main Authors: Sellars, Melony J., Trewin, Carolyn, McWilliam, Sean M., Glaves, R. S. E., Hertzler, Philip L.
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01.06.2015; Springer Nature B.V
• Subjects: Animals; Aquaculture; Automation; Biomedical and Life Sciences; Biotechnology; blastomeres; Blastomeres - cytology; Blastomeres - metabolism; cell cycle; Cell Differentiation; Cell division; complementary DNA; data collection; Databases, Genetic; Decapoda; Embryo, Nonmammalian; Embryonic Development - genetics; Embryonic growth stage; Embryos; Engineering; Expressed Sequence Tags; Female; Females; Freshwater & Marine Ecology; gastrulation; Gastrulation - genetics; gene expression; Gene Expression Regulation, Developmental; Gene Ontology; Genes; Genes, Developmental; Genetic engineering; Genomic Library; germ cells; Germ Cells - cytology; Germ Cells - metabolism; high-throughput nucleotide sequencing; Life Sciences; Male; Marsupenaeus japonicus; Mesoderm - cytology; Mesoderm - embryology; Mesoderm - metabolism; Microbiology; Molecular Sequence Annotation; ontogeny; Original Article; Penaeidae - cytology; Penaeidae - embryology; Penaeidae - metabolism; Penaeus; Phylogenetics; RNA; Seawater; Sequence Analysis, DNA; sex determination; Shellfish; shrimp; Spawning; Studies; Transcriptome; Zoology; Australia; Mesoderm; Embryo transcriptome; Kuruma shrimp; Sex determination; Germ line
• ISSN: 1436-2228; 1436-2236; 1436-2236
• DOI: 10.1007/s10126-015-9613-4

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 .