Clonal analysis of the blastoderm anlage of the Malpighian tubules in Drosophila melanogaster.

Genetically marked maroon-like (mal) clones were induced by mitotic recombination with X-rays at the blastoderm stage in mal/mal heterozygotes and were analysed in differentiated Malpighian tubules (MT). Marked cells were not confined to single anterior (MA) or posterior (MP) tubules, but were distributed among the four tubules. About 70% of the clones with two or more cells were fragmented, i.e. mal cells were separated by wild-type cells. Since the clones contain, on average, 6 cells and the differentiated MT consist of 484 cells (2 × 136 MA cells, 2 × 106 MP cells), we estimate that there are about 80 cells in the blastoderm anlage which on average pass through two to three mitoses. With increasing radiation doses (254 R, 635 R, 1270 R) a linear increase in clone frequency is observed. The mean sizes and size distributions of clones, however, remain unchanged. Since the increasing radiation dose also results in fewer differentiated Malpighi cells, we assume that regeneration does not occur. Therefore, size distributions of marked clones presumably represent real mitotic patterns in normogenesis. We suggest that essentially three successive mitoses take place, with a decreasing fraction of cells showing mitotic activity. Only a small fraction of cells goes through a fourth or even a fifth mitosis. Marked non-Minute clones induced in Minute heterozygotes are more frequent, but are not larger than non-Minute clones in wild-type background. Therefore, compartment boundaries cannot be recognized by this method. However, frequencies of marked cells found simultaneously in MA and MP pairs or in several single tubules of the same individuals are significantly higher than frequencies of multiple recombination events predicted by the Poisson distribution. From this, we conclude that neither the MA pair nor the MP pair nor single tubules represent compartments of the MT anlage.