A genomic approach to reveal novel genes associated with myotube formation in the model teleost, Takifugu rubripes
A genomic approach to reveal novel genes associated with myotube formation in the model teleost, Takifugu rubripes
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DOI：
10.1152/physiolgenomics.00087.2005
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发表时间：
2005-08-11
影响因子：
4.6
通讯作者：
Johnston, IA
中科院分区：
文献类型：
--
作者：
Fernandes, JMO;Mackenzie, MG;Johnston, IA
人们对调节脊椎动物肌管产生的转录网络知之甚少。在本研究中，我们已经使用了基因组的方法来发现新的基因与肌管形成的老虎河豚鱼，红鳍东方鲀的快肌。每个肌节的快肌纤维数量增加，直到1.2公斤体重，随后的增长是由纤维肥大单独。从180-g（肌管+）和3.4-kg（肌管-）鱼制备正向和反向消减cDNA文库，并获得1，452个表达序列标签（EST）。在这些EST被分成非冗余的簇并去除管家基因和结构基因后，选择57个基因，并使用定量PCR来研究它们在适应相同环境条件和饮食的独立组的肌管（+）和肌管（+）鱼的不同组织中的表达水平。11个新的基因被发现是一致的差异表达，但只有4个表现出适当的组织特异性表达。这四个基因在肌管（-）的快肌中相对于肌管（+）生长阶段上调5-25倍，而它们的表达在所研究的其他组织中保持不变。这些新的基因也存在于其他脊椎动物的基因组中，可能在抑制脊椎动物肌肉中肌管形成中发挥作用。
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.