The cell cycle and its relation to growth during pattern regulation in wing discs of Drosophila

Abstract

In this paper we have examined the timing of growth-rate changes, of alterations in the cell cycle, and of the reestablishment of epithelial continuity during the early stages of regulative growth of imaginal wing discs cultured in vivo. Tissue organization was disrupted prior to culture, by dissociation, and centrifugally reaggregated pellets of cells were cultured in adult female flies. Significant increases in cell numbers were detectable during the first day of culture. Flow cytometric analysis of the DNA content of cells in reaggregates during culture indicated that during the first half-day of culture, a significant transient increase in the proportions of S-phase and post-S-phase cells occurred. Scanning electron microscopic examination of dissociated cells during culture confirmed that tissue reorganization began during the first day and was nearly complete by the end of the second day. During the second day of culture, when the growth rate was maximal, the proportions of pre-( G 1 ) and post-S-phase and mitotic ( G 2 M) cells resembled those of intact premetamorphic wing discs. In contrast to disrupted tissue, intact or minimally wounded wing discs showed practically no change in cell number during culture. However, with both disrupted and intact tissue, the proportion of G 1 cells increased significantly during the culture period so that by the fourth day of culture G 1 cells predominated. Under our conditions all implants grew extensively, but regeneration by cultured reaggregates formed from pure presumptive notum and pure presumptive distal wing fragments was observed in those implants which grew the most. Our results suggest that both DNA synthesis and transient increased proportions of 4C cells may occur as an early response to injury and that these may precede the onset of cell divisions even under conditions where growth is initiated early. Our observations also suggest that the onset of cell divisions, though it occurs well before the end of the first day of culture, may nonetheless be preceded by cellular contacts and intercellular communication.