3-hydroxyacyl-coA dehydratase (HACD) enzymes are required for the elongation of fatty acids with very long carbon chain lengths (VLCFA). There are four HACD enzymes in mammals (HACD1-4), each with a distinct tissue distribution: HACD1 is specifically expressed in developing and mature striated muscles. Mutations in HACD1 cause myopathies in humans, dogs and mice, however the specific functions of this enzyme and VLCFA in muscle physiology are poorly understood. Centronuclear myopathy in Labrador dogs is the best characterised of these conditions and results in progressive disorganisation of tubuloreticular membranes and mislocalised triads. We aimed to evaluate the suitability of zebrafish for studying the functions of HACD enzymes and the role of HACD1 in muscle development and disease. Zebrafish homologues of HACD enzyme genes were identified and by conventional and quantitative RT-PCR and in situ hybridisation we characterised the expression of HACD enzymes during embryonic development and in adult tissues of AB wildtype zebrafish. Like mammals, zebrafish express two main isoforms of the HACD1 homologue of which the full length, active isoform was specifically expressed in developing and mature striated muscles. Full length hacd2 was ubiquitously expressed in adult zebrafish tissues and upregulated early in embryonic development with strong expression in the eyes and brain. hacd3 was also widely expressed, however both hacd2 and hacd3 transcripts had relatively low abundance in skeletal muscle. We documented putative zebrafish hacd4 at low levels in several tissues. CRISPR/Cas9 genome editing was used to introduce mutations into hacd1 and a muscle phenotype was observed in F0 embryos including abnormal tail morphology, motor function and disorganised myofibres with abnormal RYR1 immunostaining. In conclusion, the zebrafish homologues of the four HACD enzymes have expression and tissue distribution similar to that seen of mammals; in particular hacd1 shows strong, specific expression in developing and mature striated muscles. Mutations in hacd1 result in skeletal muscle abnormalities and reproduce key features of HACD1-CNM. This suggests that zebrafish are a good model for studying HACD enzyme functions in muscle including the pathophysiology of HACD1-deficient myopathies.