Fruitless RNAi knockdown in males interferes with copulation success in Schistocerca gregaria.
ABSTRACT In Drosophila melanogaster, the male-specific splice isoform of the fruitless gene (Fru(M)) codes for a set of transcription factors that are involved in the regulation of male courtship and copulation. Fru(M) is expressed in an interconnected neuronal circuit containing central and sensory neurons as well as motor neurons. A partial sequence from the Schistocerca gregaria fru-gene from an EST database allowed quantitative real time analysis of fru-expression in adult locusts, and revealed the highest expression in the testes, accessory glands as well as the brain (and optic lobes). Starting fru specific RNAi knockdown in the third and fourth nymphal stage resulted in a significantly lower cumulative copulation frequency of the RNAi-treated animals compared to controls after 3 h of observation. In addition, the testes of RNAi-treated males weigh less. Analysis of the egg pods resulting from a successful copulation event revealed that egg pods from females that mated with an RNAi-treated male were smaller and contained less fertilized eggs compared to egg pods from females who mated with control males. Starting injections in the fifth nymphal stage showed the complete opposite for the cumulative copulation frequency and testes weight. We conclude that already in the early nymphal phases of male desert locusts, fruitless starts to play an important role in the regulation of successful copulation in the adult. The RNAi treatment in the male has also its effects on fertility and fecundity. It remains unknown whether this effect is coming from aberrant courtship behaviour or from an altered composition of the sperm or seminal fluids.
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ABSTRACT: There has been much debate concerning whether cis-regulatory or coding changes are more likely to produce evolutionary innovation or adaptation in gene function, but an additional complication is that some genes can dramatically diverge through alternative splicing, increasing the diversity of gene function within a locus. The fruitless gene is a major transcription factor with a wide range of pleiotropic functions, including a fundamental conserved role in sexual differentiation, species-specific morphology and an important influence on male sexual behaviour. Here, we examine the structure of fruitless in multiple species of Drosophila, and determine the patterns of selective constraint acting across the coding region. We found that the pattern of selection, estimated from the ratio of non-synonymous to synonymous substitutions, varied considerably across the gene, with most regions of the gene evolutionarily conserved but with several regions showing evidence of divergence as a result of positive selection. The regions that showed evidence of positive selection were found to be localised to relatively consistent regions across multiple speciation events, and are associated with alternative splicing. Alternative splicing may thus provide a route to gene diversification in key regulatory loci.Heredity advance online publication, 23 October 2013; doi:10.1038/hdy.2013.106.Heredity 10/2013; DOI:10.1038/hdy.2013.106 · 3.80 Impact Factor
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ABSTRACT: One of the core genes in the circadian regulation network is clock (clk). By forming a heterodimer with CYCLE (CYC) that binds on an E-box in the promoter region, it induces the transcription of other elements in the circadian transcriptional feedback loops and different clock output genes. In contrast to other insects, a clk double-stranded RNA (dsRNA) treatment is lethal in adults and fifth instar nymphs of the desert locust, Schistocerca gregaria, in a dose-dependent manner. Clk knock down fifth instar nymphs are able to undergo their imaginal moult but, depending on the amount of dsRNA, it takes them longer than the controls to reach adulthood. As adults, clk knock down animals do not develop their fat body and ovaries like the control animals. Therefore, we tested the expression of different genes involved in energy metabolism and reproduction to see the effect of the clk RNA interference knock down. Surprisingly, the expression of the vitellogenin gene was up-regulated in the clk knock down females who did not appear to invest their energy in egg development. Taken together, our results point out that the clk gene in the desert locust has an additional function in development besides its established role in maintaining the circadian rhythms in the brain.Insect Molecular Biology 03/2012; 21(3):369-81. DOI:10.1111/j.1365-2583.2012.01143.x · 2.98 Impact Factor
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ABSTRACT: In all living organisms, behavior, metabolism and physiology are under the regulation of a circadian clock. The molecular machinery of this clock has been conserved throughout the animal kingdom. Besides regulating the circadian timing of a variety of processes through a central oscillating mechanism in the brain, these circadian clock genes were found to have a function in peripheral tissues in different insects. Here, we provide evidence that the circadian clock genes period (per) and timeless (tim) have a role in the male locust reproduction. A knockdown of either of the two genes has no effect on male sexual maturation or behavior, but progeny output in their untreated female copulation partners is affected. Indeed, the fertilization rates of the eggs are lower for females with a per or tim RNAi copulation partner as compared to the eggs deposited by females that mated with a control male. As the sperm content of the seminal vesicles is higher in per or tim knockdown males, we suggest that this phenotype could be caused by a disturbance of the circadian regulated sperm transfer in the male reproductive organs, or an insufficient maturation of the sperm after release from the testes.Insect biochemistry and molecular biology 12/2011; 42(2):109-15. DOI:10.1016/j.ibmb.2011.11.003 · 3.42 Impact Factor