Isoform-specific control of male neuronal differentiation and behavior in Drosophila by the fruitless gene.

BACKGROUND: How the central nervous system (CNS) develops to implement innate behaviors remains largely unknown. Drosophila male sexual behavior has long been used as a model to address this question. The male-specific products of fruitless (fru) are pivotal to the emergence of this behavior. These putative transcription factors, containing one of three alternative DNA binding domains, determine the neuronal substrates for sexual behavior in male CNS. RESULTS: We isolated the first fru coding mutation, resulting in complete loss of one isoform. At the neuronal level, this isoform alone controls differentiation of a male-specific muscle and its associated motorneuron. Conversely, a combination of isoforms is required for development of serotonergic neurons implicated in male copulatory behavior. Full development of these neurons requires the male-specific product of doublesex, a gene previously thought to act independently of fru. At the behavioral level, missing one isoform leads to diminished courtship behavior and infertility. We achieved the first rescue of a distinct fru behavioral phenotype, expressing a wild-type isoform in a defined subset of its normal expression pattern. CONCLUSION: This study exemplifies how complex behaviors can be controlled by a single locus through multiple isoforms regulating both developmental and physiological pathways in different neuronal substrates.

Functional conservation of the fruitless male sex-determination gene across 250 Myr of insect evolution.

Male sexual behavior in the fruit fly Drosophila melanogaster is regulated by fruitless (fru), a sex-determination gene specifying the synthesis of BTB-Zn finger proteins that likely function as male-specific transcriptional regulators. Expression of fru in the nervous system specifies male sexual behavior and the muscle of Lawrence (MOL), an abdominal muscle that develops in males but not in females. We have isolated the fru ortholog from the malaria mosquito Anopheles gambiae and show the gene's conserved genomic structure. We demonstrate that male-specific mosquito fru protein isoforms arise by conserved mechanisms of sex-specifically activated and alternative exon splicing. A male-determining function of mosquito fru is revealed by ectopic expression of the male mosquito isoform FRUMC in fruit flies; this results in MOL development in both fru-mutant males and fru+ females who otherwise develop no MOL. In parallel, we provide evidence of a unique feature of muscle differentiation within the fifth abdominal segment of male mosquitoes that strongly resembles the fruit fly MOL. Given these conserved features within the context of 250 Myr of evolutionary divergence between Drosophila and Anopheles, we hypothesize that fru is the prototypic gene of male sexual behavior among dipteran insects.

GAL4 enhancer trap targeting of the Drosophila sex determination gene fruitless.

The fru4 allele of the sex determination gene fruitless is induced by insertion of a P[lacZ,ry+] enhancer trap element. This insert also acts to disrupt expression of the fru P1 promoter derived male-specific proteins, consequently impairing male courtship behavior. fru4 maps less than 2 kb upstream of the fru P3 promoter, whose function is essential for viability. We replaced this insert with a GAL4 element, P[GAL4,w+], recovering two lines with insertions in opposite orientations at the locus, one of which demonstrated fru-specific mutant phenotypes. Reporter expression of these lines recapitulated that of P3- and P4-derived proteins which, when correlated with a developmental and tissue specific survey of fru promoters' activities, uncovered a previously unsuspected complexity of fru regulation. These novel fru alleles provide the tools for manipulation of fru-expressing cells, allowing the consequent effects to be related back to specific fru functions and the regulatory units controlling these activities.