Role of cell adhesion molecules during net formation in Hydorodictyon reticulatum

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A monoclonal antibody that was raised against a homogenate of zoospores recognized cell adhesion molecules localized to the contact sites between zoospores. The molecule was constantly expressed in zoospores during net formation and decreased after the completion of the adhesion between zoospores. The subcellular localization of the molecule during net formation was analyzed by indirect immunofluorescence microscopy. The peripheral regions of the spherical zoospores and the contact regions of the polyhedral zoospores were stained brightly. Immediately after cessation of the movement of polyhedral zoospores, the molecule was located on the closely apposed surfaces of two cells. In order to examine the precise spatial distribution of the molecule, zoospores and net cells were double-stained with this monoclonal antibody and a monoclonal antibody against chicken β-tubulin. The molecule closely colocalized with the microtubule bundles at the contact regions of the polyhedral zoospores and the net cells. Colchicine-treated zoospores could not form hexagonal nets, then the peripheral regions of zoospores stained uniformly. Immunoelectron microscopy revealed the presence of gold particles on a Golgi stack and the plasma membrane in polyhedral zoospores and the contact regions of net cells. I treated isolated zoospores with this monoclonal antibody. Treatment with the antibody inhibited formation of hexagonal nets. These results suggest that localization of the molecule participate in the determination of the contact sites of zoospores and that the microtubule bundles are involved in the localization of the molecule during hexagonal net formation.