Factors controlling prespore cell differentiation in Dictyostelium discoideum: minute amounts of...

Amoebae of strain V12M2 differentiate efficiently into prespore cells without cell contact in a salt solution containing cAMP, if the pH of the medium is maintained suitably acidic using a restricted buffer [31]. Under such conditions, most cells differentiate into prespore cells at pH 5.2. Using this developmental system, the elements controlling prespore-cell differentiation were analyzed. First, the dependence on cell density was examined. At a very low density (10(2) cells/cm2), most cells did not differentiate. As the density was increased the proportion of prespore cells differentiating increased, reaching a maximum at 5 x 10(3) cells/cm2. Conditioned medium could mimic the effects of cell density on cell differentiation. These findings suggest the presence of factor(s) released into the medium which are involved in inducing prespore-cell differentiation. The conditioned medium was found to contain at least two prespore-inducing components; one is a novel factor(s) and the other is DIF, which has previous only been considered to repress prespore-cell differentiation. These findings were supported by experiments using a DIF-deficient mutant, HM44.