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Ryanodine receptors (RyR) are essential regulators of cellular calcium homeostasis and signaling. Vertebrate genomes contain multiple RyR gene isoforms, expressed in different tissues and executing different functions. In contrast, invertebrate genomes contain a single RyR-encoding gene and it has long been proposed that different transcripts generated by alternative splicing may diversify their functions. Here, we analyze the expression and function of alternative exons in the  C .  elegans  RyR gene  unc-68 . We show that specific isoform subsets are created via alternative promoters and via alternative splicing in  unc-68  Divergent Region 2 (DR2), which actually corresponds to a region of high sequence variability across vertebrate isoforms. The expression of specific  unc-68  alternative exons is enriched in different tissues, such as in body wall muscle, neurons and pharyngeal muscle. In order to infer the function of specific alternative promoters and alternative exons of  unc-68 , we selectively deleted them by CRISPR/Cas9 genome editing. We evaluated pharyngeal function, as well as locomotor function in swimming and crawling with high-content computer-assisted postural and behavioral analysis. Our data provide a comprehensive map of the pleiotropic impact of isoform-specific mutations and highlight that tissue-specific  unc-68  isoforms fulfill distinct functions. As a whole, our work clarifies how the  C .  elegans  single RyR gene  unc-68  can fulfill multiple tasks through tissue-specific isoforms, and provide a solid foundation to further develop  C .  elegans  as a model to study RyR channel functions and malfunctions.

Tissue-specific isoforms of the single C. elegans Ryanodine receptor gene unc-68 control specific functions