The discovery of nitric oxide as a regulator of blood pressure, which was awarded the Nobel prize in 1995, launched intense investigation into its function in human health. Recognizing the diversity, complexity and conservation of nitric oxide actions, studies in model organisms appear relevant and needed. In our studies of Drosophila, nitric oxide induced behavioral and physiological changes consistent with a conserved role in adaptation to low oxygen (hypoxia). Seeking other parallels to its function in mammals, we found that nitric oxide activates innate immune responses in Drosophila. We developed robust assays in which tagged transgenes report immune induction in larvae or cultured cells (S2 cells) in response to nitric oxide, or to bacteria, or to hypoxia. The responses of S2 cells can be blocked by inactivation of specific genes by RNA interference (RNAi). To exploit this powerful avenue for genetic dissection, we constructed a library of 7,200 RNAs representing the conserved genes of Drosophila. We propose high-throughput RNAi screens for genes contributing to immune induction. In preliminary work, we identified the genes involved in the response to bacterial components and will do the same for genes involved in the responses to nitric oxide and hypoxia. We will further exploit our assays to define the sequence of gene action, thereby delineating distinctions and commonalities in the pathways transducing these signals. Using in vivo genetics and tests in culture, we will place the signaling pathways in their biological context. Localization of function will position gene action in a cascade that conveys immune responses from the site of infection to distant tissues. These studies will provide new models for the action of signals central to human physiology and health. ? ? ?
