The intestine is a self-renewing organ under constant stresses. Its homeostasis (controlled regulation) plays a critical role in organ function and prevention of tumor generation. Similar to humans, drosophila possess intestinal stem cells that differentiate to produce a self-renewing gut epithelium. This is an excellent model to study how cellular interactions and infection regulate tissue renewal. When drosophila midgut enterocytes (ECs) are damaged as a result of aging or stress, the ECs undergo apoptosis. This results in stimulation of midgut epithelial stem cells to divide and regenerate ECs via a poorly understood signaling pathway. In order to elucidate additional factors that may regulate this process, RNA sequencing of the drosophila midgut post stress treatment was performed. Stress induction was produced via JNK activation, apoptosis, heat shock, and treatment with paraquot and pseudomonas entomophila (P.e.). 44 genes have been shown to be up-regulated in all stress conditions when compared to a control. Among them, the eyes shut (EYS) gene is required for proper eye development in humans and flies, but no function for EYS in the intestine has been described in any model organism. In order to confirm upregulation of EYS, RNA extractions from the ECs of the midgut and qPCR experiments were performed after flies were subjected to stressors for 20 hours. Stress on EYS drosophila, including bacterial infection and oxidative stress, and subsequent dissection and RNA show an up-regulation of EYS in the midgut. Next, to test the requirement of EYS, overexpression of RNAis against EYS in drosophila ECs and subsequent quantification of stem cell proliferation via phospho-histone3 positive scoring has shown reduced stem cell proliferation in EYS depleted drosophila midgut compared to control drosophila. Thus, EYS is required in drosophila for proper regeneration and constitutes a new regulator of intestinal homeostasis.

Analysis of eyes shut (EYS) function during intestinal regeneration in Drosophila melanogaster

255th National Meeting (2018)

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