Effects of Cations on Phagocytosis in the Ciliate Pseudomicrothorax dubius1

Various cations have been examined for their effects on phagocytosis. Media with high Ca²⁺] and low K⁺] favor phagocytosis, which is inhibited in media with high K⁺], Rb⁺], or Ba²⁺] and low Ca²⁺]. Microscopical observations of inhibited cells demonstrate that swimming behavior is not modified but they cannot perform the initial step of phagocytosis, attachment to food; when Ca²⁺ is added, cells attach to and ingest food, demonstrating rapid reversal of inhibition. Attachment is shown to be a linear function of the ratio K⁺]/Ca²⁺]^1/2 or Rb⁺]/Ca²⁺]^1/2 in the medium. The Ca²⁺ influx inhibitor Verapamil blocks attachment, as does La³⁺; the latter is believed to compete with Ca²⁺ for access to the Ca²⁺ channel. Likewise, treatment of cells with media containing no added Ca²⁺ inhibits attachment, and addition of 10 μM Ca²⁺ allows 90% of these cells to attach to and ingest food. The ionophore A23187, known to transport Ca²⁺ into a wide variety of cells, provokes lysosomal streaming movements typical of attachment. Based upon these observations, Ca²⁺ influx plays an essential role in attachment; K⁺ efflux also appears to be necessary since tetraethylammonium chloride blocks attachment. Treatment of cells with Tetrodotoxin, an inhibitor of Na⁺ transport, or suspending them in media containing no added Na⁺ does not affect attachment or ingestion, indicating that Na⁺ is not implicated in these processes. An hypothesis is presented which implicates Ca²⁺ in both direct adhesion and exocytosis phenomena during attachment.