New mutants of Paramecium tetraurelia defective in a calcium control mechanism: genetic and behavioral characterizations.

The k-shy mutants of Paramecium tetraurelia are altered in several Ca2+-dependent functions which regulate ciliary motility. The isolation, genetics, and phenotypes of these mutants are described. Of six independent isolates, all contained recessive single-factor mutations and comprise two unlinked loci, ksA and ksB. All k-shy strains showed prolonged backward swimming responses to depolarizing stimuli, but gave infrequent responses to some stimuli. At least four k-shy strains displayed temperature sensitivity. Neither ksA nor ksB was allelic or linked to dancer, a mutation causing weak Ca2+ current inactivation and prolonged backward swimming. Analysis of ks+; Dn double mutants revealed synergism between the two mutations. The ksA mutant survived Ba2+ solutions longer than wild type, but was more sensitive to K+. Together with previous studies, these results are consistent with a defect in reducing intracellular Ca2+ causing both prolonged ciliary reversal and reduced Ca2+ channel activity due to more active Ca2+-dependent feedback mechanisms. The integration of the Ca2+-dependent stimulatory and inhibitory functions is therefore dependent on ks+ gene functions. The ksA mutant was rescued by microinjection of wild-type cytoplasm, suggesting a possible behavioral assay for factors related to the ksA+ gene product.