High temperature-induced sterility in the female Nile tilapia, Oreochromis niloticus.

High temperature treatments induce germ cell loss in gonads of vertebrate animals, including fish. It could be a reliable source for induction of sterility if the treatments led to a permanent loss of germ cells. Here we report that heat treatment at 37 °C for 45-60 days caused a complete loss of germ cells in female Nile tilapia, Oreochromis niloticus, and that sterility was achieved in fish at all stages of their life cycle. Unlike previous observations, germ cells did not repopulate even after returning them to the water at control conditions suggesting permanent depletion of germ cells. Gonadal somatic cells immunopositive for 3ß-hydroxysteroid dehydrogenase (3ß-HSD) were clustered at one end of the germ cell depleted gonads close to the blood vessel. Serum level of testosterone, 11-ketotestosterone, and 17ß-estradiol was significantly decreased in sterile fish compared to control. Body weight of sterile fish was higher than control fish at the end of experiment. Our observations of increased growth and permanent sterilization in the high temperature-treated fish suggest that this method could be an appropriate and eco-friendly tool for inducing sterility in fish with a higher thermal tolerance.

Identification, characterization and feeding response of peptide YYb (PYYb) gene in grass carp (Ctenopharyngodon idellus).

The peptide YYb (PYYb) is a fish-specific peptide belonging to the neuropeptide Y (NPY) family. In the present study, the full-length cDNA sequence and genomic structure of PYYb (gcPYYb) from Ctenopharyngodon idellus have been isolated and characterized. The gcPYYb gene consists of three exons interspaced by two introns, opposing to the typical architecture of most NPY-family genes as well as its paralogs. Alignment of deduced amino acid sequence indicates that the fish PYYb is more variable compared to NPY and PYY as shown by more residue changes in teleosts lineage, suggesting mild selective pressure imposed on the peptide. Real-time quantitative PCR analysis shows the gcPYYb mRNA in developing larvae is increased during the mixed endo- and exogenous-feeding period and is widely distributed throughout the intestine of fully grown individuals. Following a single meal, the gcPYYb mRNA in foregut is increased at 3 h post-feeding and subsequently decreased before the foregut contents are cleared. These results suggest that the gcPYYb has an important role in the early life stages of grass carp and is involved in food intake by transmitting feeding-related signals.

Molecular cloning, expression analysis, and potential food intake attenuation effect of peptide YY in grass carp (Ctenopharyngodon idellus).

The peptide YY (PYY) is a 36 amino acid peptide involved in the food intake control in vertebrates. We have cloned and characterized a PYY gene from grass carp Ctenopharyngodon idellus. The full-length cDNA encodes a precursor protein of grass carp PYY (gcPYY) that consists of a putative 28-amino acid signal peptide, a 36-amino acid mature peptide, an amidation-proteolytic site, and a 30-amino acid carboxy-terminal extension. The gcPYY gene is comprised of 4 exons interspaced by 3 introns as seen in PYYs from other species. Amino acid alignment and gene structure comparison indicate that the structure of PYY is well preserved throughout vertebrate phylogeny. The tissue distribution and postprandial changes in gcPYY mRNA expression were evaluated by real-time PCR, which showed that the gcPYY is expressed abundantly in the central nervous system, with significantly increased expression following a single meal. During embryogenesis, the presence of gcPYY mRNA was detected in early developing embryos, and high expression levels were observed when most larvae completed their switch from endogenous nourishment to exogenous feeding. Reduced food intake by juveniles during a single meal after giving perpheral injection of gcPYY1-36 suggests a potentially important role of PYY in the food intake attenuation in grass carp.

Molecular cloning, characterization and immunological response analysis of Toll-like receptor 21 (TLR21) gene in grass carp, Ctenopharyngodon idella.

TLR21, a non-mammalian Toll-like receptor, has been recently identified in fishes, frogs and birds. In the present study, the full-length cDNA sequence of TLR21 (CiTLR21) from Ctenopharyngodon idella has been isolated and characterized. The CiTLR21 full-length cDNA sequence consists of 3578bp, with an open reading frame (ORF) of 2958bp encoding 985 amino acid residues. The putative CiTLR21 protein contains a signal peptide sequence, 17 leucine-rich (LRR) motifs, a transmembrane region and a Toll/interleukin-1 receptor (TIR) domain. The CiTLR21 gene is expressed in a wide range of tissues with the highest expression in skin. Upon induction by Aquareovirus, CiTLR21 expression is significantly down-regulated in liver and spleen, whereas is significantly up-regulated in liver and spleen after Aeromonas hydrophila infection. These results suggest that CiTLR21 plays an important role in Aquareovirus and A. hydrophila-related diseases. This work may provide the basis for further investigations into the immune system of grass carp and other teleost fishes.

Identification of TNF13b (BAFF) gene from grass carp (Ctenopharyngodon idella) and its immune response to bacteria and virus.

The TNF superfamily B cell activating factor (BAFF) is a central cytokine in several diseases. A BAFF gene has been cloned from grass carp (Ctenopharyngodon idella), analyzed its structure, and investigated its expression pattern in various tissues after Aeromonas hydrophila and Aquareovirus infection. The open reading frame of grass carp BAFF (gcBAFF) consists of 804 bases encoding 267 amino acids. Phylogenetic analysis shows the gcBAFF is most closely related to other teleost BAFFs with the highest similarity to zebrafish. RT-PCR analysis shows the gcBAFF transcript is expressed in a wide range of tissues with the highest expression in skin and spleen. Upon induction by A. hydrophila and Aquareovirus, its expression is significantly up-regulated in gill, liver, kidney, spleen and skin as compared to PBS injected fish. The association of increased BAFF expression after bacterial and viral infections suggests that it plays a potentially important role in immune system of fish.