Female receptivity phenotype of icebox mutants caused by a mutation in the L1-type cell adhesion molecule neuroglian.

Relatively little is known about the genes and brain structures that enable virgin female Drosophila to make the decision to mate or not. Classical genetic approaches have identified several mutant females that have a reluctance-to-mate phenotype, but most of these have additional behavioral defects. However, the icebox (ibx) mutation was previously reported to lower the sexual receptivity of females, without apparently affecting any other aspect of female behavior. We have shown that the ibx mutation maps to the 7F region of the Drosophila X chromosome to form a complex complementation group with both lethal and viable alleles of neuroglian (nrg). The L1-type cell adhesion molecule encoded by nrg consists of six immunoglobulin-like domains, five fibronectin-like domains, one transmembrane domain and one alternatively spliced intracellular domain. The ibx strain has a missense mutation causing a glycine-to-arginine change at amino acid 92 in the first immunoglobulin domain of nrg. Defects in the central brain of ibx mutants are similar to those observed in another nrg mutant, central brain deranged(1) (ceb(1)). However, both ceb(1) homozygous and ceb(1)/ibx heterozygous females are receptive. The expression of a transgene containing the non-neural isoform of nrg rescues both the receptivity and the brain structure phenotypes of ibx females.

Technical knockout, a Drosophila model of mitochondrial deafness.

Mutations in mtDNA-encoded components of the mitochondrial translational apparatus are associated with diverse pathological states in humans, notably sensorineural deafness. To develop animal models of such disorders, we have manipulated the nuclear gene for mitochondrial ribosomal protein S12 in Drosophila (technical knockout, tko). The prototypic mutant tko(25t) exhibits developmental delay, bang sensitivity, impaired male courtship, and defective response to sound. On the basis of a transgenic reversion test, these phenotypes are attributable to a single substitution (L85H) at a conserved residue of the tko protein. The mutant is hypersensitive to doxycyclin, an antibiotic that selectively inhibits mitochondrial protein synthesis, and mutant larvae have greatly diminished activities of mitochondrial redox enzymes and decreased levels of mitochondrial small-subunit rRNA. A second mutation in the tko gene, Q116K, which is predicted to impair the accuracy of mitochondrial translation, results in the completely different phenotype of recessive female sterility, based on three independent transgenic insertions. We infer that the tko(25t) mutant provides a model of mitochondrial hearing impairment resulting from a quantitative deficiency of mitochondrial translational capacity.

The effects of ectopic white and transformer expression on Drosophila courtship behavior.

The sex determining genes of Drosophila males and females function to establish the potential for sex-specific behaviors. Previous studies suggest that ectopic GAL4-directed misexpression of the female-specific isoform of the sex-determining gene transformer (tra) in specific sub-domains of an otherwise male brain can lead to bisexual courtship behavior, thus identifying brain domains that may mediate sex-specific behavior. However, expression of mini-white, the marker gene used in both P[GAL4] and P[UAS(G)] constructs, also induces males to court other males, questioning whether GAL4-mediated tra expression alone can induce bisexual behavior. Here we demonstrate the consequences of inducing mutations in the mini-white genes within P[GAL4] and P[UAS(G)] constructs to generate flies in which a white mutant phenotype is revealed. In these mini-white mutant strains, P[GAL4]-mediated transformer expression alone is both sufficient and necessary to generate bisexual behavior. In addition, using RT-PCR, we reveal the presence of female transcripts of doublesex and fruitless in the brains of otherwise male (XY) flies exhibiting P[GAL4]-directed tra-expression, demonstrating that P[GAL4]-directed tra is functional at the molecular level. We conclude that P[GAL4]-directed misexpression of tra is responsible for the bisexual behavior previously described and that this is mediated via sex-specific splicing of dsx and fru. Our results support the validity of such strategies for identifying regions of the fly brain that underlie sex-specific behaviors.

Abnormal courtship conditioning in males mutant for the RI regulatory subunit of Drosophila protein kinase A.

The previously described site-selected P-element mutagenesis of a Drosophila gene encoding the regulatory subunit of cAMP-dependent protein kinase generates mutants that have defective behavior in the olfactory learning test. Here we describe the effect of the same mutations in a courtship conditioning assay. Wild-type males can distinguish between virgin females (which they court vigorously), and fertilized females (which they court less vigorously). After exposure to fertilized females, wild-type males modify their behavior by decreasing courtship to subsequent target virgins, an effect that may last for many hours. Like wild-type males, PKA-RI mutant males are also able to distinguish between virgin and fertilized females. PKA-RI males also modify their behavior towards virgin females after prior exposure to a fertilized female, but such an effect is short-lived, suggesting a defect in memory rather than learning. We also show that under these conditions the behavior of PKA-RI males is similar to that of amnesiac, dunce and rutabaga males.

Sexual behaviour: secrets and flies.

Recent studies of Drosophila courtship mutants provide a molecular foundation for sexual orientation and behaviour.

Metazoan nuclear genes for mitoribosomal protein S12.

We have characterized nuclear genes for mitoribosomal protein S12 (mt-rps12) a major component of the ribosomal accuracy centre, in human, mouse and Drosophila melanogaster. In human and Drosophila, and probably also in mouse, there is a single intron within the coding region, located in the mitochondrial targeting pre-sequence. In humans, the mRNA structure is highly suggestive of translational regulation. In all three species, there is an amino-acid substitution with respect to eubacterial homologues in a residue implicated in aminoglycoside resistance. The only viable mutant allele of the Drosophila gene, associated with a bang-sensitive phenotype (paralysis upon mechanical vibration, arising from a mechanoreceptor cell defect) also has a novel substitution in a conserved region implicated in translational fidelity. Given the involvement of the mitoribosomal accuracy centre in human sensorineural deafness by virtue of rRNA mutations, our results indicate that this fly mutant may be a useful animal model of this disorder, and earmark the gene for mt-rps12 as a candidate in human hearing impairment.

Functional dissection of the Drosophila mushroom bodies by selective feminization of genetically defined subcompartments.

Relatively little is known about the neural circuitry underlying sex-specific behaviors. We have expressed the feminizing gene transformer in genetically defined subregions of the brain of male Drosophila, and in particular within different domains of the mushroom bodies. Mushroom bodies are phylogenetically conserved insect brain centers implicated in associative learning and various other aspects of behavior. Expression of transformer in lines that mark certain subsets of mushroom body intrinsic neurons, and in a line that marks a component of the antennal lobe, causes males to exhibit nondiscriminatory sexual behavior: they court mature males in addition to females. Expression of transformer in other mushroom body domains, and in control lines, has no such effect. Our data support the view that genetically defined subsets of mushroom body intrinsic neurons perform different functional roles.

The inactive mutation leads to abnormal experience-dependent courtship modification in male Drosophila melanogaster.

Flies carrying the inactive mutation of Drosophila melanogaster have only 15% wild-type titers of the putative neurotransmitter octopamine. With a view to discovering whether the inactive mutation impairs learning, I describe the effect of the inactive mutation on experience-dependent courtship modification (EDCM). Wild-type males rapidly condition to immature males and modify their behaviour toward subsequent target flies. The weaker EDCM phenotype of inactive males is similar to that of dunce males. Using time-sampling, habituation is most rapid for wild-type males and slowest for dunce males, with inactive males showing an intermediate phenotype. These results support the notion that octopamine plays a significant role in the manifestation of Drosophila learning behavior.

The effect of the inactive mutation on longevity, sex, rhythm and resistance to p-cresol in Drosophila melanogaster.

Flies carrying the inactive (iav) mutation exhibit low locomotor activity and poor mating success, both of which are associated with a deficiency in the putative neurotransmitter, octopamine. Several other aspects of the iav mutant phenotype are described here. Male and female iav mutants show a small reduction in longevity but it is not clear whether this is a consequence of the iav mutation or their inactive phenotype. Young iav males show extended attractiveness to older courting males, which supports the notion that the iav gene has a role in post-eclosional maturation. The eclosion rhythm of iav mutants is normal, discounting the possibility of a role for octopamine in the maintenance of circadian rhythm. Flies carrying the iav mutation are highly susceptible to the octopamine analogue p-Cresol. Other phenotypically inactive flies show wild type levels of p-Cresol resistance. This is attributed to the deficiency of octopamine in iav mutants because low octopamine levels may be unable to out-compete the toxic effect of p-Cresol. Some inferences on the possible mode of action of the iav gene product are discussed.