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Molecular structure and expression patterns of flounder (Paralichthys olivaceus) Myf-5, a myogenic regulatory factor
Xungang Tan1, Yuqing Zhang, Pei-Jun Zhang
1Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, P.R. China.
Abstract:
Myf-5, a member of the myogenic regulatory factors (MRF), has been shown to be expressed in muscle precursors in early stage zebrafish embryos. The MRFs, including MyoD, Myf-5, Myogenin and MRF4, belong to the basic Helix-Loop-Helix transcription factors that contain a conserved basic Helix-Loop-Helix (bHLH) domain. To better understand the role of Myf-5 in the development of fish muscles, we have isolated the Myf-5 genomic sequence and cDNA from Flounder (Paralichthys olivaceus), and analyzed its structures and patterns of expression. Promoter analysis identified several putative transcription factor binding sites such as an E-box, NF-Y sites that might confer muscle-specific expression. Myf-5 transcripts were first detected in the paraxial mesoderm that gives rise to slow muscles. During somitogenesis, Myf-5 expression was found in developing somites. Myf-5 expression decreased gradually in somites in the anterior region, but remained strong in the newly formed somites. In the hatching stage, the expression was also detected in other muscle cells such as head muscle and fin muscle. In the growing fish, RT-PCR results showed that Myf-5 was expressed in the skeletal muscle and intestine.
Insights
This study details the Myf-5 gene
Area of Science:
- Developmental biology
- Molecular biology
- Genetics
Background:
- Myf-5 is a key myogenic regulatory factor (MRF) crucial for muscle development.
- MRFs, including Myf-5, are basic Helix-Loop-Helix transcription factors essential for muscle precursor differentiation.
- Understanding Myf-5's role in fish muscle development is vital for comparative studies.
Purpose of the Study:
- To isolate and characterize the Myf-5 gene from Flounder (Paralichthys olivaceus).
- To analyze the structure and expression patterns of Flounder Myf-5 during embryonic and larval development.
- To investigate potential regulatory elements involved in muscle-specific expression.
Main Methods:
- Genomic DNA and cDNA isolation of Flounder Myf-5.
- Bioinformatic promoter analysis to identify transcription factor binding sites.
- Whole-mount in situ hybridization and RT-PCR to determine expression patterns.
Main Results:
- The Myf-5 genomic sequence and cDNA were successfully isolated from Flounder.
- Promoter analysis revealed putative muscle-specific regulatory elements like E-box and NF-Y sites.
- Myf-5 transcripts were detected in paraxial mesoderm, somites, head muscle, and fin muscle during development.
- Expression was also observed in skeletal muscle and intestine of growing fish.
Conclusions:
- Flounder Myf-5 plays a significant role in the development of various muscle types, including skeletal, head, and fin muscles.
- The identified regulatory elements suggest conserved mechanisms for muscle-specific gene expression.
- This research provides foundational insights into the molecular basis of muscle development in teleost fish.
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