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Physiol. Genomics (May 31, 2005). doi:10.1152/physiolgenomics.00087.2005
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Submitted on April 18, 2005
Accepted on May 27, 2005

A genomic approach to reveal novel genes associated with myotube formation in the model teleost, Takifugu rubripes

Jorge M. O Fernandes1, Matthew G MacKenzie1, Greg Elgar2, Yuzuru Suzuki3, Shugo Watabe4, James R Kinghorn5, and Ian A Johnston1*

1 Gatty Marine Laboratory, School of Biology, University of St Andrews, St Andrews, Fife, United Kingdom
2 Genome Campus, Hinxton, MRC Rosalind Franklin Centre for Genomics Research, Cambridge, United Kingdom
3 Fisheries Laboratory, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Maisaka, Shizuoka, Japan
4 Laboratory of Aquatic Molecular Biology and Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan
5 Harold, Mitchell Building, School of Biology, University of St Andrews, St Andrews, Fife, United Kingdom

* To whom correspondence should be addressed. E-mail: iaj{at}st-and.ac.uk.

Little is known about the transcriptional networks that regulate myotube production in vertebrates. In the present study, we have used a genomic approach to discover novel genes associated with myotube formation in fast muscle of the tiger puffer fish, Takifugu rubripes. The number of fast muscle fibers per myotome increased until 1.2 kg body mass, and subsequent growth was by fiber hypertrophy alone. Forward and reverse subtracted cDNA libraries were prepared from a 180 g (myotube +) and a 3.4 kg (myotube -) fish and 1,452 expressed sequence tags (ESTs) were obtained. After grouping these ESTs into non-redundant clusters and eliminating housekeeping and structural genes, 57 genes were selected and qPCR was used to investigate their expression levels in different tissues from independent groups of myotube (-) and myotube (+) fish acclimated to the same environmental conditions and diet. Eleven novel genes were found to be consistently differentially expressed, but only 4 showed appropriate tissue-specific expression. These 4 genes were up-regulated 5 to 25 times in fast muscle of myotube (-) relative to myotube (+) growth stages, whilst their expression remained unchanged in the other tissues studied. The novel genes identified, which are also present in other vertebrate genomes, may play a role in inhibiting myotube formation in vertebrate muscle.




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