Analysis of RNA Expression in Adult Zebrafish Skeletal Muscle

  • Tamar E. SztalEmail author
  • Peter D. Currie
  • Robert J. Bryson-Richardson
Part of the Methods in Molecular Biology book series (MIMB, volume 1668)


The zebrafish is an excellent vertebrate model system to investigate skeletal muscle development and disease. During early muscle formation the small size of the developing zebrafish allows for the characterization of gene expression in whole embryos. However, as the zebrafish develops, access to the underlying skeletal muscle is limited, requiring the skeletal muscle to be sectioned for a more detailed examination. Here, we describe a straightforward and effective method to prepare adult zebrafish skeletal muscle sections, preserving muscle morphology, to characterize gene expression in the zebrafish adult skeletal muscle.

Key words

Zebrafish adult skeletal muscle Pathology Vibratome sectioning RNA in situ hybridization Skeletal muscle 



The research was funded by an Australian National Health and Medical Research Council (NHMRC) Project Grant (APP1010110). TES is supported by MDA Developmental Grant (APP381325) and AFM Telethon Postdoctoral Fellowship (APP19853). PDC is supported by NHMRC Principal Research Fellowships (APP1002147, APP1041885).


  1. 1.
    Nowak KJ, Ravenscroft G, Laing NG (2012) Skeletal muscle α-actin diseases (actinopathies): pathology and mechanisms. Acta Neuropathol 125:19–32CrossRefPubMedGoogle Scholar
  2. 2.
    Joyce NC, Oskarsson B, Jin L-W (2012) Muscle biopsy evaluation in neuromuscular disorders. Phys Med Rehabil Clin N Am 23:609–631CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Bönnemann CG, Wang CH, Quijano-Roy S et al (2014) Diagnostic approach to the congenital muscular dystrophies. Neuromuscul Disord 24(4):289–311CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Fetterman GH, Wratney MJ, Donaldson JS, Danowski TS (1956) Muscular dystrophy. I. History, clinical status, muscle strength, and biopsy findings. AMA J Dis Child 91:326–338CrossRefPubMedGoogle Scholar
  5. 5.
    Dubowitz V, Sewry C, Oldfors A, Lane R (2007) Histological and histochemical changes. In: Muscle biopsy: a practical approach, 4th edn. Sauders Elsevier, pp 55–95Google Scholar
  6. 6.
    Kumar A, Accorsi A, Rhee Y, Girgenrath M (2015) Do's and don'ts in the preparation of muscle cryosections for histological analysis. J Vis Exp 99:e52793Google Scholar
  7. 7.
    Sheriffs IN, Rampling D, Smith VV (2001) Paraffin wax embedded muscle is suitable for the diagnosis of muscular dystrophy. J Clin Pathol 54:517–520CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Shi S-R, Liu C, Pootrakul L et al (2008) Evaluation of the value of frozen tissue section used as “gold standard” for immunohistochemistry. Am J Clin Pathol 129:358–366CrossRefPubMedGoogle Scholar
  9. 9.
    Meng H, Janssen PML, Grange RW et al (2014) Tissue triage and freezing for models of skeletal muscle disease. J Vis Exp 89:e51586Google Scholar
  10. 10.
    Chatterjee S (2014) Artefacts in histopathology. J Oral Maxillofac Pathol 18:S111–S116CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Sztal T, Berger S, Currie PD et al (2011) Characterization of the laminin gene family and evolution in zebrafish. Dev Dyn 240:422–431CrossRefPubMedGoogle Scholar
  12. 12.
    Broadbent J, Read EM (1999) Wholemount in situ hybridization of Xenopus and zebrafish embryos. Methods Mol Biol 127:57–67CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Tamar E. Sztal
    • 1
    Email author
  • Peter D. Currie
    • 2
  • Robert J. Bryson-Richardson
    • 1
  1. 1.School of Biological SciencesMonash UniversityMelbourneAustralia
  2. 2.Australian Regenerative Medicine InstituteMonash UniversityMelbourneAustralia

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