Involvement of actin and myosins in Plasmodium berghei ookinete motility.

Ookinetes of the genus Plasmodium are motile, invasive cells that develop in the mosquito midgut following ingestion of a parasite-infected blood meal. We show here that ookinetes display gliding motility on glass slides in the presence of insect cells. Moreover, in addition to stationary "flexing" and "twirling" of the cells, two distinct types of movements occur: productive forward translocational motility in straight segment that progresses with an average speed of approximately 6mum/min and rotational motility, which does not lead to forward translocation. Locomotion is reduced by treatment with butanedione monoxime, an inhibitor of myosin ATPase, and by three different actin inhibitors. We also studied the expression during ookinete development of genes encoding actin and two small class XIV myosins, PbMyoA, and PbMyoB. Western immunoblots revealed that PbMyoA is only present in fully mature ookinetes, whilst the other two proteins are additionally expressed in gametocytes and zygotes. Immunofluorescence experiments reveal that MyoA and actin co-localize in the apical tip of the parasite whereas MyoB displays a punctate pattern of expression around the entire cell periphery. Following treatment with jasplakinolide, the apparent level of detectable actin appears to substantially increase and becomes concentrated in a discrete area in the basal pole of the ookinete.