Mutation of cfap58 causes sperm malformation and reduced fertility in zebrafish

Mature sperms play a crucial role in fish reproduction, directly affecting offspring survival and quality. Cilia- and flagella-associated protein 58 ( cfap58 ), localized to motile cilia, is essential for sperm flagellar assembly and male fertility. Defects of CFAP58 lead to severe multiple morphological abnormalities of the flagella (MMAF), exhibiting sperm flagellar malformations and motility impairments in mammals. However, current studies have mostly focused on mammals, with little known about fish. Here, we used zebrafish as a model organism to study the effect of cfap58 on fish fertility. Sperm carrying cfap58 mutations generated via CRISPR/Cas9 technology exhibited significantly reduced fertilization rate and embryo viability. Mutant sperm had abnormal flagella morphology, with the conserved “9 + 2” microtubule structure disrupted, abnormal mitochondrial morphology, and significantly reduced sperm motility. We also observed a significant increase in the proportion of undifferentiated germ cells in cfap58 −/− testis. Single cell transcriptome sequencing of testis revealed that cfap58 −/− showed extensive dysregulation of genes involved in multiple cilia and microtubule related pathways. Our data suggest that cfap58 plays an important role in sperm maturation and fertility maintenance in zebrafish. Additionally, we propose that cfap58 may participate in the composition of radial spokes and contribute to maintaining the normal function of intraflagellar transport (IFT) within the sperm flagellum.