From an immunologist perspective, sharks are an important group of jawed cartilaginous fishes and survey of the public database revealed a great gap in availability of large-scale sequence data for the group of Chondrichthyans the elasmobranchs. In an attempt to bridge this deficit we generated the transcriptome from the spleen and kidney tissues (a total of 1,606,172 transcripts) of the shark, Chiloscyllium griseum using the Illumina HiSeq2000 platform. With a cut off of > = 300 bp and an expression value of >1RPKM we used 43,385 transcripts for BLASTX analysis which revealed 17,548 transcripts matching to the NCBI nr database with an E-value of < = 10−5 and similarity score of 40%. The longest transcript was 16,974 bases with matched to HECT domain containing E3 ubiqutin protein ligase. MEGAN4 annotation pipeline revealed immune and signalling pathways including cell adhesion molecules, cytokine-cytokine receptor interaction, T-cell receptor signalling pathway and chemokine signaling pathway to be highly expressed in spleen, while different metabolism pathways such as amino acid metabolism, carbohydrate metabolism, lipid metabolism and xenobiotic biodegradation were highly expressed in kidney. Few of the candidate genes were selected to analyze their expression levels in various tissues by real-time PCR and also localization of a receptor by in-situ PCR to validate the prediction. We also predicted the domains structures of some of the identified pattern recognition receptors, their phylogenetic relationship with lower and higher vertebrates and the complete downstream signaling mediators of classical dsRNA signaling pathway. The generated transcriptome will be a valuable resource to further genetic and genomic research in elasmobranchs.
Toda buffalo is a native genetically isolated semi‐wild breed, maintained by the aboriginal Toda tribes, present in the Nilgiri Hills of Tamil Nadu, India. The available population is only 1800. It is a brown medium sized animal with long horns set wide apart in a circular fashion. This breed resembles the swamp buffaloes, but based on karyological studies, it is classified under the river buffalo. The expression profile of toll‐like receptor (TLR) 1–10 mRNAs in spleen, mesenteric lymph node, lung, skin, jejunum and rectum were assessed using TaqMan real time PCR with GAPDH as endogenous control from Toda and Murrah buffaloes (n=4). The Ct values for different TLR mRNA in the tissues were compared among each breed with spleen expression levels as calibrator as well as between the two breeds and analyzed using the relative expression software tool (REST 2008‐V2.0.7). The Toda buffaloes showed higher expression levels of TLR 1 and 10 mRNA in the lymph node, TLR 1 in jejunum and TLR 1, 5, 6, 7 and 10 mRNAs in the skin. However, in Murrah buffaloes, lowered expression levels of TLR 10 mRNA in lung, TLR 4 in lymph node and TLR 10 in rectum and skin were observed. The Toda buffaloes showed significantly higher levels of TLR9 mRNA in the lymph node and TLR 1,5,6,9 and 10 mRNAs in the skin than Murrah. Thus, higher expression levels of a wide repertoire of TLR mRNA were detected in the skin of Toda buffaloes. The resistance of the Toda buffalo to protozoan parasites such as Theileria or Babesia, as is known anecdotally, could be related to the stronger innate immune responses mediated by TLRs in the skin.This work has been funded under the National Agricultural Innovative Project, Indian Council of Agricultural Research, New Delhi
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