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Dead-End (dnd) Gene Cloning and Gonad-Specific Expression Pattern in Starry Flounder (Platichthys stellatus).

ABSTRACT: dnd is a germline-specific maternal RNA expressed in various vertebrate classes, which encodes an RNA-binding protein that is essential for PGC migration. The purpose of this study is fundamental research about starry flounder dnd gene for germ cell marker development. In this study, we cloned and analyzed the expression levels of Platichthys stellatus dead end (psdnd) in various tissues and embryonic stages. The psdnd gene was isolated from starry flounder ovaries, cloned into a pGEM-t vector, and sequenced. Full-length of psdnd cDNA was 1495 bp long, encoding 395 amino acids. psdnd expression levels were investigated by real-time polymerase chain reaction (qRT-PCR) in various tissues and embryo developmental stages. psdnd transcripts were detected in the testes and ovaries, but not in somatic tissues. Embryonic psdnd expression levels were higher during early embryo development stages than during late embryogenesis; psdnd expression was highest at the 1 cell stage, then gradually decreased throughout the subsequent developmental stages. The spatial expression pattern was analyzed by whole-mount in situ hybridization (WISH). The psdnd transcripts migration pattern was similar with zebrafish (Danio rerio). Our results suggest that psdnd may function as a germ cell-specific marker.

SUBMITTER: Yoon JH

PROVIDER: S-EPMC8388513 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:A feeding trial was conducted to assess the feasibility of supplementing taurine in soy-based diets for juvenile starry flounder Platichthys stellatus. The basal diet (Crude protein 66.5%, crude lipid 8.5%) was supplemented with 0 (control), 0.5%, 1.0%, 1.5%, 2.0% and 2.5% taurine to formulate six test diets. Each diet was fed to 40 juvenile fish (22.25 g) in triplicate tanks (120 L) attached to a sea water circulation-system. Fish were fed twice daily by hand to apparent satiation during the 56-d trial. At the end of the trial, fish were counted and weighed for the analyses of growth performance, diet utilization and survival after a 24-h fast. Blood, intestines and muscles were collected for the analyses of serum oxidation resistance, digestive enzymes and body compostion. Livers were collected from the remaining fish at 4 h post-feeding for metabolic enzymes analyses. The results showed that fish fed diets supplemented with 1.0-2.5% taurine grew from 22.25-22.26 g to 47.88-50.40 g with higher average weight gain (25.62-28.12 vs 23.07 g ), specific growth rate (1.37-1.46 vs 1.27%/d ), feed intake (1.04-1.06 vs 1.00%/d), protein efficiency (2.50-2.61 vs 2.44) and lower feed conversion rate (0.84-0.83 vs 0.89) than the control treatment. Diets supplemented with 1.5-2.5% taurine significantly elevated the activities of pepsin (2.47-2.55 vs 2.22, U mg-1 prot), trypsin of distal intestine(14.55-15.24 vs 11.94, U mg-1 prot), hepatic glucokinase (126.62-129.42 vs 105.56, U mg-1 prot) and fatty acid synthetase (125.56-136.89 vs 108.45, U mg-1 prot). All diets supplemented with taurine increased the activities of lipase (32.23-36.67 vs 29.53, U g-1 prot) and trypsin (35.85-37.89 vs 33.54, U mg-1 prot) of proximal intestine, hepatic aspartate transaminase (736.990-832.38 vs 699.24, U mg-1 prot), alanine aminotransferase (477.40-551.86 vs 373.97, U mg-1 prot) and glycogen synthase (2.16-2.59 vs 1.97, U mg-1 prot), as well as serum superoxide dismutase (4.33-4.59 vs 4.07, U mg-1 prot ) and glutathione peroxidase (42.23-50.25 vs 39.17, mol mg-1 prot). Therefore, taurine supplementation benefits juvenile starry flounder growth, digestion, nutrients metabolism and oxidation resistance. The optimal taurine requirement for starry flounder is 1.75%, and the recommended supplementation level is at least 1.6% for maximizing growth of fish fed a low-fishmeal diet (13.6%).

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Dead-End (dnd) Gene Cloning and Gonad-Specific Expression Pattern in Starry Flounder (Platichthys stellatus).

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