Title: A shotgun approach to identify mechanical nociception genes Short title: Screen for mechanical nociception genes

CC-BY-NC-ND 4.0 International license peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was not. Abstract: The molecular mechanisms of sensing noxious mechanical force by nociceptive sensory neurons remain poorly understood. Traditional methods for probing mechanical nociception behavioral responses are labor intensive and involve the testing of one animal at a time. This time consuming process of behavioral testing has largely precluded large scale analyses. Indeed, large scale genetic screens that have been performed thus far have been largely restricted to the investigation of ion channel genes [1]. Here we describe a new behavioral assay for mechanical nociception in which tens of animals can be stimulated simultaneously. In this assay, third instar larvae of the genetically tractable organism Drosophila melanogaster are mechanically stimulated with tungsten particles that are fired from a gun. We have used the new assay to carry out a genetic screen in which we investigated the function of 231 nociceptor enriched genes with tissue-specific RNA interference. Targeting of 21 genes resulted in mechanically insensitive phenotypes and targeting of a single gene resulted in a hypersensitive mechanical nociception phenotype. 16 of the 22 mechanical nociception genes have clearly identified mammalian homologs and 10 of these homologs show enriched expression in mammalian nociceptors. Six of the identified genes were previously uncharacterized and these were named after famed Roman gladiators