Mutants in Drosophila TRPC Channels Reduce Olfactory Sensitivity to Carbon Dioxide

Background

Members of the canonical Transient Receptor Potential (TRPC) class of cationic channels function downstream of Gαq and PLCβ in Drosophila photoreceptors for transducing visual stimuli. Gαq has recently been implicated in olfactory sensing of carbon dioxide (CO₂) and other odorants. Here we investigated the role of PLCβ and TRPC channels for sensing CO₂ in Drosophila.

Methodology/Principal Findings

Through behavioral assays it was demonstrated that Drosophila mutants for plc21c, trp and trpl have a reduced sensitivity for CO₂. Immuno-histochemical staining for TRP, TRPL and TRPγ indicates that all three channels are expressed in Drosophila antennae including the sensory neurons that express CO₂ receptors. Electrophysiological recordings obtained from the antennae of protein null alleles of TRP (trp³⁴³) and TRPL (trpl³⁰²), showed that the sensory response to multiple concentrations of CO₂ was reduced. However, trpl³⁰²; trp³⁴³ double mutants still have a residual response to CO₂. Down-regulation of TRPC channels specifically in CO₂ sensing olfactory neurons reduced the response to CO₂ and this reduction was obtained even upon down-regulation of the TRPCs in adult olfactory sensory neurons. Thus the reduced response to CO₂ obtained from the antennae of TRPC RNAi strains is not due to a developmental defect.

Conclusion

These observations show that reduction in TRPC channel function significantly reduces the sensitivity of the olfactory response to CO₂ concentrations of 5% or less in adult Drosophila. It is possible that the CO₂ receptors Gr63a and Gr21a activate the TRPC channels through Gαq and PLC21C.     

Extrachromosomal Inheritance of Carbon Dioxide Sensitivity in the Mosquito CULEX QUINQUEFASCIATUS

Mosquitoes from a laboratory colony of Culex quinquefasciatus from Matsu Island, China, develop irreversible paralytic symptoms after exposure to carbon dioxide at 1 degree. This CO2 sensitivity is caused by an inherited infectious agent, probably a virus. Crossing studies between CO2-sensitive and -resistant mosquitoes showed that the sensitivity trait is inherited extrachromosomally in a fashion strictly analogous to the hereditary transmission of sigma virus in Drosophila melanogaster. Sensitivity could be maintained through maternal transmission alone, despite nine generations of backcrossing of "stabilized" CO2-sensitive females to males from a resistant strain. CO2-sensitive males crossed to resistant females transmitted sensitivity to a portion of their F1 progeny, and only the female F1 sensitives were capable of further hereditary transmission.--Matsu, or a very similar hereditary infectious agent, is common in natural populations of Cx. quinquefasciatus on Oahu, Hawaii. Fifty-nine percent of the families reared from field-collected egg rafts contained CO2-sensitive mosquitoes, and some families contained only sensitive mosquitoes.     

Interactions of Carbon Dioxide and Food Odours in Drosophila: Olfactory Hedonics and Sensory Neuron Properties

Behavioural responses of animals to volatiles in their environment are generally dependent on context. Most natural odours are mixtures of components that can each induce different behaviours when presented on their own. We have investigated how a complex of two olfactory stimuli is evaluated by Drosophila flies in a free-flying two-trap choice assay and how these stimuli are encoded in olfactory receptor neurons. We first observed that volatiles from apple cider vinegar attracted flies while carbon dioxide (CO₂) was avoided, confirming their inherent positive and negative values. In contradiction with previous results obtained from walking flies in a four-field olfactometer, in the present assay the addition of CO₂ to vinegar increased rather than decreased the attractiveness of vinegar. This effect was female-specific even though males and females responded similarly to CO₂ and vinegar on their own. To test whether the female-specific behavioural response to the mixture correlated with a sexual dimorphism at the peripheral level we recorded from olfactory receptor neurons stimulated with vinegar, CO₂ and their combination. Responses to vinegar were obtained from three neuron classes, two of them housed with the CO₂-responsive neuron in ab1 sensilla. Sensitivity of these neurons to both CO₂ and vinegar per se did not differ between males and females and responses from female neurons did not change when CO₂ and vinegar were presented simultaneously. We also found that CO₂-sensitive neurons are particularly well adapted to respond rapidly to small concentration changes irrespective of background CO₂ levels. The ability to encode temporal properties of stimulations differs considerably between CO₂- and vinegar-sensitive neurons. These properties may have important implications for in-flight navigation when rapid responses to fragmented odour plumes are crucial to locate odour sources. However, the flies’ sex-specific response to the CO₂-vinegar combination and the context-dependent hedonics most likely originate from central rather than peripheral processing.     

Selection on X-Linked Genes during Speciation in the Drosophila Athabasca Complex

We present the results of a restriction site survey of variation at five loci in Drosophila athabasca, complimenting a previous study of the period locus. There is considerably greater differentiation between the three semispecies of D. athabasca at the period locus and two other X-linked genes (no-on-transient-A and E74A) than at three autosomal genes (Xdh, Adh and RC98). Using a modification of the HKA test, which uses fixed differences between the semispecies and a test based on differences in Fst among loci, we show that the greater differentiation of the X-linked loci compared with the autosomal loci is inconsistent with a neutral model of molecular evolution. We explore several evolutionary scenarios by computer simulation, including differential migration of X and autosomal genes, very low levels of migration among the semispecies, selective sweeps, and background selection, and conclude that X-linked selective sweeps in at least two of the semispecies are the best explanation for the data. This evidence that natural selection acted on the X-chromosome suggests that another X-linked trait, mating song differences among the semispecies, may have been the target of selection.     

Carbon Dioxide and Fruiting in Sphaerobolus

In cultures bathed in air deprived of its CO² fruiting in Sphaerobolusis greatly reduced or fails completely. However, increasingthe concentration of CO² in the air to 1or 5 per cent tendsto reduce fruiting as compared with normal air. The stimulatoryeffect of CO² in normal air appears to be exerted at the stageof development when sporophore primordia are being produced.The CO² does not seem to require the immediate presence of lightto exert its effect. It has not been found possible to replacethe effect of CO² by substituting Krebs-cycle organic acids.Using ¹⁴CO² it is shown that ¹⁴C is incorporated in the myceliumboth in light and in darkness.     

Short- and Long-Term Inhibition of Respiratory Carbon Dioxide Efflux by Elevated Carbon Dioxide

Dark carbon dioxide efflux rates of recently fully expandedleaves and whole plants of Amaranthus hypochondriacus L., Glycinemax (L.) Merr., and Lycopersicon esculentum Mill. grown in controlledenvironments at 35 and 70 Pa carbon dioxide pressure were measuredat 35 and 70 Pa carbon dioxide pressure. Harvest data and whole-plant24-h carbon dioxide exchange were used to determine relativegrowth rates, net assimilation rates, leaf area ratios, andthe ratio of respiration to photosynthesis under the growthconditions. Biomass at a given time after planting was greaterat the higher carbon dioxide pressure in G. max and L. esculentum,but not the C₄ species, A. hypochondriacus. Relative growthrates for the same range of masses were not different betweencarbon dioxide treatments in the two C₃ species, because highernet assimilation rates at the higher carbon dioxide pressurewere offset by lower leaf area ratios. Whole plant carbon dioxideefflux rates per unit of mass were lower in plants grown andmeasured at the higher carbon dioxide pressure in both G. maxand L. esculentum, and were also smaller in relation to daytimenet carbon dioxide influx. Short-term responses of respirationrate to carbon dioxide pressure were found in all species, withcarbon dioxide efflux rates of leaves and whole plants lowerwhen measured at higher carbon dioxide pressure in almost allcases. Amaranthus hypochondriacus L., Glycine max L. Merr., Lycopersicon esculentum Mill., soybean, tomato, carbon dioxide, respiration, growth