The TOR and Insulin/IGF signalling (IIS) network controls growth, metabolism and ageing. Although reducing TOR or insulin signalling can be beneficial for ageing, it can be detrimental for wound healing, but the reasons for this difference are unknown. Here we show that IIS is activated in the cells surrounding an epidermal wound in Drosophila melanogaster larvae, resulting in PI3K activation and redistribution of the transcription factor FOXO. Insulin and TOR signalling are independently necessary for normal wound healing, with FOXO and S6K as their respective effectors. IIS is specifically required in cells surrounding the wound, and the effect is independent of glycogen metabolism. Insulin signalling is needed for the efficient assembly of an actomyosin cable around the wound, and constitutively active myosin II regulatory light chain suppresses the effects of reduced IIS. These findings may have implications for the role of insulin signalling and FOXO activation in diabetic wound healing.

Cell–cell adhesion and cell shape are regulated at adherens junctions during embryonic morphogenesis. Beati et al. show that the Drosophila LIM domain protein Smallish interacts with Bazooka, Canoe, and Src42A at adherens junctions. Loss-of-function and gain-of-function phenotypes reveal a function for Smallish in regulation of actomyosin contractility and cell shape.

The Drosophila larva has been used to investigate many processes in cell biology, including morphogenesis, physiology, responses to drugs and new therapeutic compounds. Despite its enormous potential as a model system, it has technical limitations in cases where longer-term live imaging is necessary, because of the lack of efficient methods for immobilising larvae for extended periods. We describe here a simple procedure for anaesthetisation and long-term in vivo imaging of the epidermis and other larval organs including gut, imaginal discs, neurons, fat body, tracheae and haemocytes, and show a procedure for probing cell properties by laser ablation. We include a survey of different anaesthetics, showing that short exposure to diethyl ether is the most effective for long-term immobilisation of larvae. This method does not require specific expertise beyond basic Drosophila genetics and husbandry, and confocal microscopy. It enables high-resolution studies of many systemic and subcellular processes in larvae.

Epithelial wound healing in Drosophila involves the formation of multinucleate cells surrounding the wound. We show that autophagy, a cellular degradation process often deployed in stress responses, is required for the formation of a multinucleated syncytium during wound healing, and that autophagosomes that appear near the wound edge acquire plasma membrane markers. In addition, uncontrolled autophagy in the unwounded epidermis leads to the degradation of endo‐membranes and the lateral plasma membrane, while apical and basal membranes and epithelial barrier function remain intact. Proper functioning of TORC1 is needed to prevent destruction of the larval epidermis by autophagy, in a process that depends on phagophore initiation and expansion but does not require autophagosomes fusion with lysosomes. Autophagy induction can also affect other sub‐cellular membranes, as shown by its suppression of experimentally induced laminopathy‐like nuclear defects. Our findings reveal a function for TORC1‐mediated regulation of autophagy in maintaining membrane integrity and homeostasis in the epidermis and during wound healing.

No abstract

Mutations in the hedgehog pathway drive the formation of tumors in many different organs, including the development of basal cell carcinoma in the skin. However, little is known about the role of epidermal Indian hedgehog (Ihh) in skin physiology. Using mouse genetics, we identified overlapping and distinct functions of Ihh in different models of epidermal tumorigenesis. Epidermal deletion of Ihh resulted in increased formation of benign squamous papilloma. Strikingly, Ihh-deficient mice showed an increase in malignant squamous cell carcinoma and developed lung and lymph node metastases. In a sebaceous gland tumor model, Ihh deficiency inhibited tumor cell differentiation. More mechanistically, IHH stimulated cell proliferation by activating the transcription factor GLI2 in human keratinocytes and human tumors. Thus, our results uncover important functions for Ihh signaling in controlling proliferation, differentiation, malignant progression, and metastasis of epithelial cancer, establishing Ihh as a gatekeeper for controlling the grade of tumor malignancy.

The Drosophila larva has been used to investigate many processes in cell biology, including morphogenesis, physiology, responses to drugs and new therapeutic compounds. Despite its enormous potential as a model system, it has technical limitations in cases where longer-term live imaging is necessary, because of the lack of efficient methods for immobilising larvae for extended periods. We describe here a simple procedure for anaesthetisation and long-term in vivo imaging of the epidermis and other larval organs including gut, imaginal discs, neurons, fat body, tracheae and haemocytes, and show a procedure for probing cell properties by laser ablation. We include a survey of different anaesthetics, showing that short exposure to diethyl ether is the most effective for long-term immobilisation of larvae. This method does not require specific expertise beyond basic Drosophila genetics and husbandry, and confocal microscopy. It enables high-resolution studies of many systemic and subcellular processes in larvae.

Epithelial wound healing in Drosophila involves the formation of multinucleate cells surrounding the wound. We show that autophagy, a cellular degradation process often deployed in stress responses, is required for the formation of a multinucleated syncytium during wound healing. In addition, uncontrolled autophagy in the unwounded epidermis leads to the degradation of endo-membranes and the lateral plasma membrane, while the apical and basal membranes and the epithelial barrier function remain intact. Proper functioning of TORC1 is needed to prevent autophagy from destroying the larval epidermis, which depends on membrane isolation and phagophore expansion, but does not require the fusion of autophagosomes to lysosomes. Our findings reveal a function for TORC1-mediated regulation of autophagy in maintaining membrane integrity and homeostasis in the epidermis and during wound healing. Finally, autophagy can counteract experimentally induced nuclear defects resembling laminopathies.