Transcriptome responses in the rectal gland of fed and fasted spiny dogfish shark (Squalus acanthias) determined by suppression subtractive hybridization

Deck, Courtney A., McKay, Sheldon J., Fiedler, Tristan J., LeMoine, Christophe M. R., Kajimura, Makiko, Nawata, C. Michele, Wood, Chris M., Walsh, Patrick J. (October 2013) Transcriptome responses in the rectal gland of fed and fasted spiny dogfish shark (Squalus acanthias) determined by suppression subtractive hybridization. Comparative Biochemistry and Physiology Part D: Genomics and Proteomics. ISSN 1744-117X

Abstract

Abstract Prior studies of the elasmobranch rectal gland have demonstrated that feeding induces profound and rapid up regulation of the gland’s ability to secrete concentrated NaCl solutions and the metabolic capacity to support this highly ATP consuming process. We undertook the current study to attempt to determine the degree to which up regulation of mRNA transcription was involved in the gland’s activation. cDNA libraries were created from mRNA isolated from rectal glands of fasted (7 days post-feeding) and fed (6 h and 22 h post-feeding) spiny dogfish sharks (Squalus acanthias), and the libraries were subjected to suppression subtractive hybridization (SSH) analysis. Quantitative real time PCR (qPCR) was also used to ascertain the mRNA expression of several genes revealed by the SSH analysis. In total the treatments changed the abundance of 170 transcripts, with 103 up regulated by feeding, and 67 up regulated by fasting. While many of the changes took place in ‘expected’ Gene Ontology (GO) categories (e.g., metabolism, transport, structural proteins, DNA and RNA turnover, etc.), KEGG analysis revealed a number of categories which identify oxidative stress as a topic of interest for the gland. GO analysis also revealed that branched chain essential amino acids (e.g., valine, leucine, isoleucine) as potential metabolic fuels for the rectal gland. In addition, up regulation of transcripts for many genes in the anticipated GO categories did not agree (i.e., fastingdown regulated in feeding treatments) with previously observed increases in their respective proteins/enzyme activities. These results suggest an ‘anticipatory’ storage of selected mRNAs which presumably supports the rapid translation of proteins upon feeding activation of the gland.

Item Type: Paper
Uncontrolled Keywords: elasmobranchs rectal gland transcript regulation feeding ion transport metabolism
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > transcriptomes
CSHL Authors:
Communities: Dolan DNA Learning Center
Depositing User: Matt Covey
Date: 6 October 2013
Date Deposited: 15 Oct 2013 20:20
Last Modified: 15 Oct 2013 20:20
URI: https://repository.cshl.edu/id/eprint/28651

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