An electrophysiological study of the regulation of ciliary beating frequency in Paramecium.

Abstract

The role of the surface membrane in the control of ciliary beat frequency in Paramecium was examined by intracellular electrophysiological techniques and pressure injection of Ca2+ and EGTA [ethylene glycol bis (.beta.-amino-ethyl ether) tetracetic acid]. Experiments were done on wild type P. caudatum and on both the wild type and a pawn mutant of P. tetraurelia. The increased frequency of beating that accompanies reversal of power stroke orientation in response to depolarization in the wild type fails to occur during depolarization in the mutant pawn, which fails to exhibit ciliary reversal upon depolarization. Injection of moderate amounts of EGTA blocked the frequency increase without interfering with reversal of the beat in response to depolarization of the wild type. Larger injection of EGTA also prevented reversed beating. The beat frequency in the normal (forward-swimming) direction increased during hyperpolarization in pawn. The hyperpolarizing frequency-voltage relations were quantitatively similar to those of the wild type. Injection of EGTA to a final concentration of 10 mM into wild type cells neither modified the resting frequency nor blocked the frequency increase with normally accompanied hyperpolarization. Transient ciliary reversal in both pawn and wild type produced by injection of Ca2+ could be terminated by the passage of inward current. The power stroke returned to the normal forward-swimming direction and the ciliary beating frequency increased. Upon termination of the inward current the cilia of Ca2+-injected cells again beat in reverse for many seconds. The results support previous reports that increased frequency of beating and ciliary reversal seen in response to depolarization both require the entry of Ca2+ through the surface membrane. Conversely, frequency increase with hyperpolarization seems independent of an altered rate of Ca2+ entry. Increased frequency during hyperpolarization appears related more closely to electrotonic membrane current than to membrane potential. Probably, inward current activates high frequency beating by altering the ionic environment of the axoneme within the restricted volume of the cilium by electrophoretic means.