Mechanisms of Horizontal Cell-to-Cell Transfer of Wolbachia spp. in Drosophila melanogaster

Abstract

<i>Wolbachia</i> is an intracellular endosymbiont present in most arthropod and filarial nematode species. Transmission between hosts is primarily vertical, taking place exclusively through the female germ line, although horizontal transmission has also been documented. The results of several studies indicate that <i>Wolbachia</i> spp. can undergo transfer between somatic and germ line cells during nematode development and in adult flies. However, the mechanisms underlying horizontal cell-to-cell transfer remain largely unexplored. Here, we establish a tractable system for probing horizontal transfer of <i>Wolbachia</i> cells between <i>Drosophila melanogaster</i> cells in culture using fluorescence <i>in situ</i> hybridization (FISH). First, we show that horizontal transfer is independent of cell-to-cell contact and can efficiently take place through the culture medium within hours. Further, we demonstrate that efficient transfer utilizes host cell phagocytic and clathrin/dynamin-dependent endocytic machinery. Lastly, we provide evidence that this process is conserved between species, showing that horizontal transfer from mosquito to <i>Drosophila</i> cells takes place in a similar fashion. Altogether, our results indicate that <i>Wolbachia</i> utilizes host internalization machinery during infection, and this mechanism is conserved across insect species.<b>IMPORTANCE</b> Our work has broad implications for the control and treatment of tropical diseases. <i>Wolbachia</i> can confer resistance against a variety of human pathogens in mosquito vectors. Elucidating the mechanisms of horizontal transfer will be useful for efforts to more efficiently infect nonnatural insect hosts with <i>Wolbachia</i> as a biological control agent. Further, as <i>Wolbachia</i> is essential for the survival of filarial nematodes, understanding horizontal transfer might provide new approaches to treating human infections by targeting <i>Wolbachia</i> Finally, this work provides a key first step toward the genetic manipulation of <i>Wolbachia</i>.