Fish Physiology and Biochemistry

, Volume 44, Issue 2, pp 679–691 | Cite as

Promoter analysis of the fish gene of slow/cardiac-type myosin heavy chain implicated in specification of muscle fiber types

  • Shigeharu KinoshitaEmail author
  • Saltuk Bugrahan Ceyhun
  • Asaduzzamann Md
  • Bhuiyan Sharmin Siddique
  • Dadasaheb B. Akolkar
  • Shuichi Asakawa
  • Shugo Watabe


Vertebrate skeletal muscles consist of heterogeneous tissues containing various types of muscle fibers, where specification of the fiber type is crucial for muscle development. Fish are an attractive experimental model to study the mechanisms of such fiber type specification because of the separated localization of slow and fast muscles in the trunk myotome. We examined regulation of expression of the torafugu gene of slow/cardiac-type myosin heavy chain, MYH M5 , and isolated an operational promoter in order to force its tissue-specific expression across different fish species via the transgenic approach in zebrafish and medaka. This promoter activity was observed in adaxial cell-derived superficial slow muscle fibers under the control of a hedgehog signal. We also uncovered coordinated expression of MYH M5 and Sox6b, which is an important transcriptional repressor for specification of muscle fiber types and participates in hedgehog signaling. Sequence comparison in the 5′-flanking region identified three conserved regions, CSR1–CSR3, between torafugu MYH M5 and its zebrafish ortholog. Analysis of deletion mutants showed that CSR1 significantly stimulates gene expression in slow muscle fibers. In contrast, deletion of CSR3 resulted in ectopic expression of a reporter gene in fast muscle fibers. CSR3 was found to contain a putative Sox family protein-binding site. These results indicate that the dual mechanism causing inhibition in fast muscle fibers and activation in slow muscle fibers is essential for slow muscle fiber-specific gene expression in fish.


Promoter analysis 



We express our sincere thanks to T. Koda and T. Mochizuki for collecting torafugu embryos and adult specimens. This study was partly supported by a Grant-in-Aid for Scientific Research from the Japan Society for Promotion of Science.

Supplementary material

10695_2018_463_MOESM1_ESM.pdf (133 kb)
ESM 1 (PDF 133 kb)


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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Shigeharu Kinoshita
    • 1
    Email author
  • Saltuk Bugrahan Ceyhun
    • 2
  • Asaduzzamann Md
    • 1
  • Bhuiyan Sharmin Siddique
    • 1
  • Dadasaheb B. Akolkar
    • 1
  • Shuichi Asakawa
    • 1
  • Shugo Watabe
    • 3
  1. 1.Department of Aquatic Bioscience, Graduate School of Agricultural and Life SciencesThe University of TokyoTokyoJapan
  2. 2.Fisheries FacultyAtaturk UniversityErzurumTurkey
  3. 3.School of Marine BiosciencesKitasato UniversitySagamiharaJapan

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