Kinetics of adhesion and de-adhesion of Chlamydomonas gametes.

In medium with low nitrogen content, vegetative strains of the unicellular biflagellate alga Chlamydomonas reinhardi form gametes. Mating type plus (mt+) and mating type minus (mt-) gametes adhere via their flagella to give aggregates in which the gametes eventually fuse to form zygotes. A quantitative assay has now been developed which measures aggregation and fusion by use of a Coulter electronic particle counter to determine loss of single gametes as they form aggregates in suspension. Determination of the rate and extent of cell fusion by microscopy agrees with the results obtained with the more rapid and convenient Coulter counter assay. By use of the assay it was found that aggregation and fusion occur at the same rate and to the same extent at 12 degrees C and 25 degrees C. Flagella from one of the mating types can specifically substitute for the corresponding live gametes; more than 70% of the gametes were aggregated and the extent of aggregation was proportional to the number of flagella added, until the ratio of cells to flagella exceeded 2. At 22 degrees C, in the flagella/gamete mixtures, adhesion was complete in less than 5 min, but at 5 to 10 min, gametes began to de-adhere from the clusters and, depending on the number of flagella added, essentially all of the gametes detached from the aggregates in 10 to 50 min. The gametes in such mixtures were fully competent to aggregate again, whereas the flagella recovered from such mixtures were shown by use of a radioactive flagella-binding assay to be inactive with fresh gametes. Inactivation of the flagella was temperature-dependent, was not catalyzed by soluble factors, and required adhesion of flagella to gametes of the opposite mating type. The potential physiological functions of the de-adhesion process are discussed.