Cholecystokinin receptors in Atlantic salmon: molecular cloning, gene expression, and structural basis.

The peptide hormone cholecystokinin (CCK) exerts a wide range of digestive and CNS-related physiological signaling via CCK receptors in brain and gut. There is very limited information available on these receptors in Atlantic salmon. The aim of this study was to characterize CCK receptors in gut and brain of salmon. We have identified and cloned one CCK-1 receptor and duplicates of CCK-2 receptor in salmon. The phylogenetic analysis indicates the existence of one common ancestor gene for all CCK receptors. CCK-1R mRNA is highly expressed in pancreas followed by midgut, hindgut, gallbladder, and stomach indicating an involvement in pancreatic regulation and gallbladder contractions. CCK-2R1/gastrin mRNA is expressed at high levels in midgut and at relatively low levels in stomach, gallbladder, and pancreas. We postulate CCK-2R1/gastrin receptor to have gastrin-related functions because of its distribution and abundance in gastro-intestinal (GI) tissues. CCK-2R2 is relatively abundant in brain but has low expression levels in gut tissues supporting the hypothesis for involvement in the gut-brain signaling. Major functional motifs and ligand interaction sites in salmon are conserved with that of mammals. This information will be instrumental for comparative studies and further targeting receptor activation and selectivity of biological responses of CCK in salmon.

Molecular cloning and genomic characterization of novel leptin-like genes in salmonids provide new insight into the evolution of the Leptin gene family.

In the current study we describe the identification of novel leptin B homologous gene/s in the four salmonid species Atlantic salmon (Salmo salar), rainbow trout (Oncorhynchus mykiss), brown trout (Salmo trutta) and Arctic charr (Salvelinus alpinus). Homology modeling of Salmo salar (Ss) LepB1/B2 suggests that the protein satisfies parameters as long-chain four helical cytokine family and that the basic structural pattern of the protein follows that of human leptin (Zhang et al., 1997). Importantly, the docking studies suggested the SsLepB has binding affinity to the AA residues that identify the leptin binding and FNIII domains of the SsLep receptor (Rønnestad et al., 2010). Phylogenetic analyses support that LepB paralogs have most probably originated by 4R whole genome duplication (WGD) before speciation of the salmonid lineages. LepB1 and LepB2 genes are both present in the two closest relatives, the Atlantic salmon and the brown trout, while rainbow trout and charr have only preserved the long LepB1 variant in their genome. We have defined the sites of SsLepB mRNA expression at key life stages in Atlantic salmon and found that SsLepB1 and SsLepB2, although to different extent, were expressed in redundant and mostly complementary fashion in brain and gills throughout the lifecycle, suggesting that this pair of paralogs is likely undergoing early stages of subfunctionalization. Furthermore, we have quantified the expression profiles of SsLepB genes and of other two recently duplicated salmon leptins (SsLepA1, SsLepA2) during early development and show evidence that in fish, as in mammals and amphibians, leptin could play important roles in growth and development. This study provides an essential groundwork to further elucidate structural and functional evolution of this important hormone in salmonids as well as in other teleosts.

Role of vitamin C on lead acetate induced spermatogenesis in swiss mice.

In the present study significantly increased lipid peroxidation value (LPP) after a single intraperitioneal injection of lead acetate (LA) (100 mg/kg b.w.) indicated enormous generation of Reactive Oxygen Species (ROS). Lead-induced ROS has a direct inhibitory effect on the growth and differentiation of the spermatogonial cells showing a significant decline in sperm count. Chromosomal analysis of the primary spermatocytes at week 4 post-treatment in lead-treated mice revealed significantly higher no of aberrant cells including chromosomal deficiency, autosomal and XY-asynapsis plates compared to untreated control mice, Sperm morphology studies at week 1-4 and at week 8 post-treatment, indicated higher percentage of deformed sperm population compared to vehicle injected groups of mice. Supplementation of vitamin C (Vit C) at a dose of 10 mg/kg body weight to lead-treated mice groups, however, significantly reduced the LPP with a concomitant increase in sperm count, marked decrease in the no of aberrant cells and significant decline in the percentage of morphologically abnormal sperm population. Protective role of Vit C in combating lead-induced oxidative stress in mice testicular cells, has been discussed.