Damage to the Drosophila follicle cell epithelium produces “false clones” with apparent polarity phenotypes

The Drosophila follicular epithelium, which surrounds developing egg chambers, is a well-established model for studying epithelial polarity because it is continuously generated from adult stem cells, making it easy to generate homozygous mutant clones in a heterozygous background. Mutant clones are usually marked by the loss of Green Fluorescent Protein (GFP) expression, which distinguishes them from their green, wild-type neighbours. Here we report that damage to the epithelium during dissection can produce groups of GFP-negative cells that resemble mutant clones. Furthermore, several polarity factors, such as aPKC and Discs large, are not localised in these damage-induced false clones. This phenotype is identical to that reported for several mutants, including ampk and Dystroglycan mutant clones under conditions of energetic stress. Using more reliable systems to mark ampk and Dystroglycan null clones such as the MARCM system, we found that neither protein is required for epithelial polarity under low energy conditions. Thus, our previous report of a specific low energy polarity pathway is an artefact of the increased damage caused by dissecting the small ovaries of starved flies. However, ampk mutant cells are larger than normal under both starvation and well-fed conditions, indicating that AMPK restricts follicle cell growth even when dietary sugar is not limiting. We suspect that several other reports of mutants that disrupt follicle cell polarity may also be based on the phenotype of damage-induced false clones, and recommend the use of positively marked clones to avoid this potential artefact.