Niflumic acid disrupts marine spermatozoan chemotaxis without impairing the spatiotemporal detection of chemoattractant gradients

Abstract

In many broadcast-spawning marine organisms, oocytes release chemicals that guide conspecific spermatozoa towards their source through chemotaxis. In the sea urchin Lytechinus pictus, the chemoattractant peptide speract triggers a train of fluctuations of intracellular Ca2+ concentration in the sperm flagella. Each transient Ca2+ elevation leads to a momentary increase in flagellar bending asymmetry, known as a chemotactic turn. Furthermore, chemotaxis requires a precise spatiotemporal coordination between the Ca2+-dependent turns and the form of chemoattractant gradient. Spermatozoa that display Ca2+-dependent turns while swimming down the chemoattractant gradient, and conversely suppress turning events while swimming up gradient, successfully approach the center of the gradient. Previous experiments in Strongylocentrotus purpuratus sea urchin spermatozoa showed that niflumic acid (NFA), an inhibitor of several ion channels, drastically altered the speract-induced Ca2+ fluctuations and swimming patterns. In this study, mathematical modeling of the speract-dependent Ca2+ signaling pathway suggests that NFA, by potentially affecting HCN, CaCC and CaKC channels, may alter the temporal organization of Ca2+ fluctuations, and therefore disrupt chemotaxis. Here we investigate our hypothesis using a novel automated method for analyzing sperm behavior. We show that NFA does indeed disrupt chemotactic responses of L. pictus spermatozoa, although the temporal coordination between the Ca2+-dependent turns and the form of chemoattractant gradient is unaltered. Instead, NFA disrupts sperm chemotaxis by altering the arc length traveled during each chemotactic turning event. This alteration in the chemotactic turn trajectory disorientates spermatozoa at the termination of the turning event. We conclude that NFA disrupts chemotaxis without affecting how the spermatozoa decode environmental cues.