Localization of glycogen synthetase during differentiation of presumptive cell types in Dictyostelium discoideum

Abstract

Ultramicrochemical techniques were utilized to assay glycogen synthetase (EC 2.4.1.11) activity in cell samples of Dictyostelium discoideum as small as 0.01 mug (dry weight) in reaction volumes of 0.1 mul. The activity was assayed by an amplification procedure employing the enzymatic cycling of pyridine nucleotides. These techniques were used to determine the extent of localization of glycogen synthetase in the two cell types during differentiation of D. discoideum. Localization studies in developing spore cells revealed decreasing enzyme activity to the culmination stage. During this phase of development, the enzyme required the presence of soluble glycogen for activity. From culmination to sorocarp stage, enzyme activity increased and was independent of the soluble glycogen. In developing stalk cells, synthetase showed a decreasing gradient of activity. In sorocarps, the cells in the stalk apex showed synthetase activity similar to that of the spores. The cells at the bottom of the stalk had no detectable activity.