Project description:Expression of known and predicted genes in tissues of Tetraodon nigroviridis (pufferfish) pooled from multiple healthy individuals.
Project description:Expression of known and predicted genes in tissues of Tetraodon nigroviridis (pufferfish) pooled from multiple healthy individuals. Two-colour experiments with two different tissues hybridized to each array. Each tissue is arrayed in replicate with dye swaps. Tissues: Beak, Brain, Calvaria, Connective tissue, Eye, Fin, Gallbladder, Gill, Heart, Intestine, Kidney, Liver, Ovary, Red muscle, Skin, Spleen, Stomach, Swimbladder, Testis, White muscle
Project description:The effects of phoenixin-14 on the hypothalamic-pituitary-gonadal (HPG) axis and spawning in green-spotted puffer (Dichotomyctere nigroviridis)
Project description:Tetraodon embryos were obtained by in vitro fertilization of eggs and their development was observed through brightfield microscopy. The embryo development was divided into distinct stages using morphological features as described for normal development of other fish species like medaka (Oryzias latipes), zebrafish (Danio rerio), and fugu (Takifugu rubripes). Total RNA was extracted with Trizol (Invitrogen) according to the manufacturer s protocol from eggs, whole embryo at 30% epiboly (30 epi) and whole embryo at 24 hours post fertilisation (24 hpf). The RNA samples were treated with 2U Dnase I (Qiagen) per μg RNA sample at 37°C for 10 minutes. Digested samples were then treated with 20 mg/mL proteinase K (Sigma Aldrich) at 37°C for 45 minutes. The quality and quantity of total RNA were assessed with the Bioanalyzer 2100 (Agilent) and no sign of degradation was detected (RIN > 9.0). Sequencing libraries were generated from total RNA samples following the Truseq RNA protocol (Illumina). Single end reads (1 x 50 nucleotides) were obtained from 3 lanes on a Hiseq1000 using SBS v3 kits (Illumina).
Project description:BackgroundThe G-protein-coupled receptors (GPCRs) constitute one of the largest and most ancient superfamilies of membrane proteins. They play a central role in physiological processes affecting almost all aspects of the life cycle of an organism. Availability of the complete sets of putative members of a family from diverse species provides the basis for cross genome comparative studies.ResultsWe have defined the repertoire of GPCR superfamily of Tetraodon complement with the availability of complete sequence of the freshwater puffer fish Tetraodon nigroviridis. Almost all 466 Tetraodon GPCRs (Tnig-GPCRs) identified had a clear human homologue. 189 putative human and Tetraodon GPCR orthologous pairs could be identified. Tetraodon GPCRs are classified into five GRAFS families, by phylogenetic analysis, concurrent with human GPCR classification.ConclusionDirect comparison of GPCRs in Tetraodon and human genomes displays a high level of orthology and supports large-scale gene duplications in Tetraodon. Examples of lineage specific gene expansions were also observed in opsin and odorant receptors. The human and Tetraodon GPCR sequences are analogous in terms of GPCR subfamilies but display disproportionate numbers of receptors at the subfamily level. The teleost genome with its expanded set of GPCRs provides additional and interesting comparators to study both evolution and function of these receptors.
