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

¹ Gatty Marine Laboratory, School of Biology, University of St. Andrews, St. Andrews, Fife, United Kingdom
² Medical Research Council Rosalind Franklin Centre for Genomics Research, Genome Campus, Hinxton, 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 Sir Harold Mitchell Building, School of Biology, University of St. Andrews, St. Andrews, Fife, United Kingdom

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 these ESTs were grouped into nonredundant clusters and housekeeping and structural genes were eliminated, 57 genes were selected and quantitative PCR 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 four showed appropriate tissue-specific expression. These four genes were upregulated 5–25 times in fast muscle of myotube(–) relative to myotube(+) growth stages, while 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.

expressed sequence tags; suppression subtractive hybridization; subtracted cDNA library; myogenesis

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