Lidocaine reversibly inhibits fertilization in Chlamydomonas: a possible role for calcium in sexual signalling

Abstract

A flagellar adhesion-induced signal sent during the mating reaction of the biflagellate alga, Chlamydomonas reinhardtii, initiates release of cell-wall-degrading enzymes, activation of mating structures, and cell fusion. The nature of this signal is unknown, but it may be mediated by an adhesion-induced change (activation) of flagellar tips. The studies reported here show that lidocaine, a local anesthetic that is reported to interfere with the movement of divalent cations across cell membranes, reversibly blocks cell wall loss and gametic fusion without blocking adhesion or flagellar tip activation. In these experiments lidocaine inhibited both the initial rates and the extent of wall loss and zygote formation. Studies with gametes of a paralyzed flagellar mutant, pf 17, revealed that lidocaine also blocked flagellar surface motility (visualized as movement of polystyrene beads) at concentrations of the inhibitor which also prevented gametic fusion. The concentration of lidocaine required to block cell fusion was dependent on the concentration of calcium or magnesium in the medium. In the absence of added calcium, 0.5 mM lidocaine inhibited fusion by 70%. In 0.5 mM calcium, 0.5 mM lidocaine had no effect on fusion and 2 mM lidocaine was required for 90% inhibition. The results suggest that divalent cations may play a critical role in sexual signalling in Chlamydomonas.