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  • Background: In Drosophila, male flies require the expression of the male-specific Fruitless protein (FRU[superscript M]) within the developing pupal and adult nervous system in order to produce male courtship and copulation behaviors. Recent evidence has shown that specific subsets of FRU[superscript M] neurons are necessary for particular steps of courtship and copulation. In these neurons, FRU[superscript M] function has been shown to be important for determining sex-specific neuronal characteristics, such as neurotransmitter profile and morphology. Results: We identified a small cohort of FRU[superscript M] interneurons in the brain and ventral nerve cord by their co-expression with the transcription factor Engrailed (En). We used an En-GAL4 driver to express a fru[superscript M] RNAi construct in order to selectively deplete FRU[superscript M] in these En/FRU[superscript M] co-expressing neurons. In courtship and copulation tests, these males performed male courtship at wild-type levels but were frequently sterile. Sterility was a behavioral phenotype as these En-fru[superscript M]RNAi males were less able to convert a copulation attempt into a stable copulation, or did not maintain copulation for long enough to transfer sperm and/or seminal fluid. Conclusions: We have identified a population of interneurons necessary for successful copulation in Drosophila. These data confirm a model in which subsets of FRU[superscript M] neurons participate in independent neuronal circuits necessary for individual steps of male behavior. In addition, we have determined that these neurons in wild-type males have homologues in females and fru mutants, with similar placement, projection patterns, and neurochemical profiles.
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