Calcium control of ciliary arrest in mussel gill cells

After several hours in 20 mM sodium phosphate and 40 mM KC1 (pH 7.4) or similar simple solutions, ciliated cells exfoliate en masse from stripped gill epithelium of freshwater mussels, e.g., Elliptio complanatus. Three types of ciliated ceils-lateral (L), laterofrontal (LF), and frontal (F)-can be distinguished and counted separately in the suspensions. About one-half of the cells of each type remain motile. Motility is unaffected by addition of 10^-5 M A23187 or 10^-2 M Ca ⁺² added separately, but when ionophore and Ca⁺² are added together, ciliary beat is largely arrested. Treatment of the cells with Triton X-100 (Rohm & Haas Co., Philadelphia, Pa.) results in a total loss of motility as the ciliary membrane becomes disrupted. Such models can be reactivated by addition of ATP and Mg⁺². All ciliated cell types are reactivated to about the same extent. At least 80 of the activity of the untreated preparation returns. Ca⁺²-EGTA buffers added to the reactivating solutions permit titration of free Ca⁺² concentration vs. percent motility. Activity is unchanged for all cell types at Ca⁺² < 10^-7 M; at 10^-6 Ca⁺², L cilia are arrested differentially, whereas at Ca⁺² > 10^-4 M most cilia of all cell types are arrested. We conclude: (a) that increasing cytoplasmic Ca⁺² is directly responsible for ciliary arrest, (b) that the readily reversible physiological arrest response of the L cilia in the intact gill is caused by a rise in free Ca⁺² in narrow limits from ca. 5 x 10^-7 M to ca. 8 x 10^-7 M, and (c) that the site which is sensitive to Ca⁺² is part of the ciliary axoneme or the basal apparatus.