Identifying downstream targets of ankrd1a in skeletal muscle injury repair in adult zebrafish

The ANKRD1 gene is responsive to different forms of mechanical stress, including injury, stretching, resistance exercise, and eccentric contractions. We showed that ankrd1a, zebrafish homologue of mammalian ANKRD1, gets activated in the heart, after cryoinjury of the ventricle and in skeletal muscle after stab wound, suggesting its role in muscle healing processes. To identify targets of ankrd1a involved in skeletal muscle repair we performed RNA-seq of injured adult ankrd1a mutant and wt zebrafish muscle at 5 days post injury. Non-injured skeletal muscle of both genotypes were used as controls. The loss of ankrd1a function affected the expression of genes involved in muscle contraction, muscle cell differentiation, MAPK and integrin-mediated signaling pathways, and cell-substrate adhesion. Our findings offer novel insights into the ankrd1a function and skeletal muscle repair in adult zebrafish. Overall design: The ankrd1abns502 mutant zebrafish line was generated by CRISPR/Cas9 genome editing. Muscle injury was performed by needle stab into the dorsal myotome of the tail. After 5 days of recovery, RNA was isolated from the muscle tissue around the injury, anterior and posterior to the needle lesion or corresponding non-injured region in control fish. Four experimental groups (injured wt, non-injured wt, injured mutant, and non-injured mutant) were subjected to RNA-seq. Each experimental group comprised four biological samples and one sample contained RNA isolated from the skeletal muscle of one fish.