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.     

Dendritic Integration in Olfactory Sensory Neurons: A Steady-State Analysis of How the Neuron Structure and Neuron Environment Influence the Coding of Odor Intensity

Response properties of the receptor potential at steady state were analyzed in a biophysical model of an olfactory sensory neuron embedded in a multicell environment. The neuron structure was described as a set of several identical dendrites (or cilia) bearing the transduction mechanisms, joined to a nonsensory part—dendritic knob, soma, and axon. The different ionic compositions of the media surrounding the neuron sensory and nonsensory parts and the extraneuronal voltage sources, which both result from the presence of auxiliary cells, were also taken into account. Analytical solutions were found to describe how the receptor potential at the nonsensory part responds to a uniform change in the odorant-dependent conductance resulting from odorant stimulation of the sensory dendrites. We investigated the influence of various geometrical and electrical parameters on the receptor-potential response in the classical model neuron within a homogeneous environment and in the model neuron surrounded with auxiliary cells. First, it was found that the maximum amplitude of the receptor potential is independent of the neuron structure in the absence of auxiliary cells but not in their presence. In the latter case, the amplitude decreases with the length and number of sensory dendrites and with the input resistance of the nonsensory part. Second, the sensitivity (as measured by the increase in membrane conductance at half-maximum response) of the neuron model in the absence of auxiliary cells is higher, but its dynamic range is narrower than in their presence. The dynamic range is wide and the sensitivity low when the input resistance of the nonsensory part is small and the sensory dendrite is unbranched. Both sensitivity and dynamic range are higher for a longer dendrite. These results help understand the morphology of insect olfactory sensilla and can be generalized to other neuron types.     

Embryonic Origin of Olfactory Circuitry in Drosophila: Contact and Activity-Mediated Interactions Pattern Connectivity in the Antennal Lobe

Olfactory neuropiles across different phyla organize into glomerular structures where afferents from a single olfactory receptor class synapse with uniglomerular projecting interneurons. In adult Drosophila, olfactory projection interneurons, partially instructed by the larval olfactory system laid down during embryogenesis, pattern the developing antennal lobe prior to the ingrowth of afferents. In vertebrates it is the afferents that initiate and regulate the development of the first olfactory neuropile. Here we investigate for the first time the embryonic assembly of the Drosophila olfactory network. We use dye injection and genetic labelling to show that during embryogenesis, afferent ingrowth pioneers the development of the olfactory lobe. With a combination of laser ablation experiments and electrophysiological recording from living embryos, we show that olfactory lobe development depends sequentially on contact-mediated and activity-dependent interactions and reveal an unpredicted degree of similarity between the olfactory system development of vertebrates and that of the Drosophila embryo. Our electrophysiological investigation is also the first systematic study of the onset and developmental maturation of normal patterns of spontaneous activity in olfactory sensory neurons, and we uncover some of the mechanisms regulating its dynamics. We find that as development proceeds, activity patterns change, in a way that favours information transfer, and that this change is in part driven by the expression of olfactory receptors. Our findings show an unexpected similarity between the early development of olfactory networks in Drosophila and vertebrates and demonstrate developmental mechanisms that can lead to an improved coding capacity in olfactory neurons.     

Formation of an Olfactory Search Image for Explosives Odours in Sniffer Dogs*

The term ‘search image’ refers to an animal's heightened ability to detect a specific cryptic prey, after experience with that prey. Formulated in 1960 by Tinbergen (Archives Neerlandaises de zoologie, vol. 13, pp. 265–343), the search image concept has been the subject of much research, albeit almost entirely based on visual search cues in birds of prey. Given the theoretical and practical importance of this concept for foraging or searching in general, we set out to investigate whether dogs could form olfactory search images for explosive odours. Seven experienced explosives detector dogs were first tested for their ability to detect 30 g of the explosives 2,4,6‐trinitrotoluene (TNT), RDX + polyisobutylene + di(2‐ethylhexyl)sebacate + fuel oil (C4) or pentaerythritol tetranitrate (PETN) placed in various containers randomly distributed along a 300‐m limestone track. In consecutive stages of the experiment, we altered the relative percentages of the three explosives. The results showed that the percentage of TNT containers detected by the dogs increased in trials that followed placing of a high percentage of TNT relative to other explosives and decreased in trials that follwed placing of a relatively low percentage of TNT containers. The percentage detection of C4 and PETN was not influenced by their relative abundance at any stage. Overall, these results suggest that dogs can form an olfactory search image that might aid in prey/target detection.     

Interactions Between Transcutaneous Ion Transfer Processes and Carbon Dioxide Excretion in Amphibians

SYNOPSIS. The amphibian skin possesses a wide variety of physiologicalfunctions in that it constitutes not only the major organ forrespiratory CO₂ exchange but also plays important roles forionic as well as osmotic balance. Apart from the simple transcutaneousdiffusion of CO₂ down its partial pressure gradient, acid-baserelevant ion exchange mechanisms in the skin may also be importantin overall pH regulation in these animals. The skin of somefrogs, for example, contains mechanisms for the exchange ofNa^$/H^$ and HCO₃^–/Cl^– in which NaCl is actively transportedinto the animal in exchange for H^$ and HCO₃^–. While suchexchange mechanisms have often been studied in the context ofosmoregulation in freshwater environments, their potential importancein acid-base regulation have been largely unexplored. The presentpaper reviews the evidence for participation of cutaneous iontransfer mechanisms in the overall regulation of CO₂ excretionand acid-base balance in amphibians.     

Notch Is Required in Adult Drosophila Sensory Neurons for Morphological and Functional Plasticity of the Olfactory Circuit

Olfactory receptor neurons (ORNs) convey odor information to the central brain, but like other sensory neurons were thought to play a passive role in memory formation and storage. Here we show that Notch, part of an evolutionarily conserved intercellular signaling pathway, is required in adult Drosophila ORNs for the structural and functional plasticity of olfactory glomeruli that is induced by chronic odor exposure. Specifically, we show that Notch activity in ORNs is necessary for the odor specific increase in the volume of glomeruli that occurs as a consequence of prolonged odor exposure. Calcium imaging experiments indicate that Notch in ORNs is also required for the chronic odor induced changes in the physiology of ORNs and the ensuing changes in the physiological response of their second order projection neurons (PNs). We further show that Notch in ORNs acts by both canonical cleavage-dependent and non-canonical cleavage-independent pathways. The Notch ligand Delta (Dl) in PNs switches the balance between the pathways. These data define a circuit whereby, in conjunction with odor, N activity in the periphery regulates the activity of neurons in the central brain and Dl in the central brain regulates N activity in the periphery. Our work highlights the importance of experience dependent plasticity at the first olfactory synapse.     

CO2与养分交互作用对番茄幼苗根生长的影响

在营养液栽培条件下,以番茄（合作906）为研究材料,设计不同的CO2及养分浓度处理,采用定期取样的方法研究番茄定植后根重齄的动态变化及CO2与不同养分供应强度的交互作用对根中碳氮含量与碳氮比的影响。结果表明：番茄苗期根干物质在生长前期积累速率较慢,中后期积累速率较快,在育苗后期CO2对根干物质积累的影响大于前期,根干重对CO2的响应随营养液离子浓度的改变而变化,表明对天番茄幼苗根的生长发育,CO2施肥结合高养分浓度的营养液,才能达到最佳效果。定量分析番茄根干鲜重与生长时间的关系,结果表明：生长条件的改变,会改变番茄根系的生长,对于根鲜重．在较高的CO2条件下,1／2山崎番茄营养液与1／4山崎番茄营养液里生长的根系鲜重在拟合方程中以幂函数拟合得到的相关系数最大,其余处理以二次曲线方程拟合得到的相关系数最大;对于根干重,在较高的CO2条件下,1／2山崎番茄营养液里生长的根系干重在所拟合的方程中以幂函数拟合的相关系数最大,其余的处理以二次曲线方程拟合的相关系数最大。CO2降低了在1／2山崎番茄营养液中生长的根系中的N含量,升高其它营养液处理中的根的N含量,降低了在1／2、1／4山崎番茄营养液中生长的番茄根系中Ｃ的百分含量,增加在1／8、1／16山崎番茄营养液中生长的番茄根系中Ｃ的百分含量,增加所有营养液浓度条件下的Ｃ、N总量,降低根系中的Ｃ／N比,在同一CO2条件下C／N比随营养液浓度的降低而升高。     

Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites,Tracheae, and Muscles in the Drosophila Larva Body Wall

To understand how differences in complex cell shapes are achieved, it is important to accurately follow microtubule organization. The Drosophila larval body wall contains several cell types that are models to study cell and tissue morphogenesis. For example tracheae are used to examine tube morphogenesis¹, and the dendritic arborization (DA) sensory neurons of the Drosophila larva have become a primary system for the elucidation of general and neuron-class-specific mechanisms of dendritic differentiation^2-5 and degeneration⁶.The shape of dendrite branches can vary significantly between neuron classes, and even among different branches of a single neuron^7,8. Genetic studies in DA neurons suggest that differential cytoskeletal organization can underlie morphological differences in dendritic branch shape^4,9-11. We provide a robust immunological labeling method to assay in vivo microtubule organization in DA sensory neuron dendrite arbor (Figures 1, 2, Movie 1). This protocol illustrates the dissection and immunostaining of first instar larva, a stage when active sensory neuron dendrite outgrowth and branching organization is occurring ^12,13.In addition to staining sensory neurons, this method achieves robust labeling of microtubule organization in muscles (Movies 2, 3), trachea (Figure 3, Movie 3), and other body wall tissues. It is valuable for investigators wishing to analyze microtubule organization in situ in the body wall when investigating mechanisms that control tissue and cell shape.Download video file.^{(47M, mov)}     

Food Preference and Appetite after Switching between Sweet and Savoury Odours in Women

Background

Exposure to food odours increases the appetite for congruent foods and decreases the appetite for incongruent foods. However, the effect of exposure to a variety of food odours, as often occurs in daily life, is unknown.

Objective

Investigate how switching between sweet and savoury odours affects the appetite for sweet and savoury products.

Design

Thirty women (age: 18-45y; BMI: 18.5-25kg/m²) intensely smelled the contents of cups filled with banana, meat or water (no-odour) in a within-subject design with four combinations: no-odour/banana, no-odour/meat, meat/banana and banana/meat. Participants received one combination per test day. In each combination, two cups with different fillings were smelled for five minutes after each other. Treatment order was balanced as much as possible. The effects of previous exposure and current odour on the appetite for (in)congruent sweet and savoury products, and odour pleasantness were analysed. A change from meat to banana odour or banana to meat odour was referred to as switch, whereas a change from no-odour to meat odour or no-odour to banana odour was no-switch.

Results

The current odour (P<0.001), as opposed to the previous exposure (P = 0.71), determined the appetite for (in)congruent sweet and savoury products, already one minute after a switch between sweet and savoury odours. The pleasantness of the odour decreased during odour exposure (P = 0.005).

Conclusions

After a switch, the appetite for specific products quickly adjusted to the new odour and followed the typical pattern as found during odour exposure in previous studies. Interestingly, the appetite for the smelled food remained elevated during odour exposure, known as sensory-specific appetite, whereas the pleasantness of the odour decreased over time, previously termed olfactory sensory-specific satiety. This seeming contradiction may result from different mechanisms underlying the odour-induced anticipation of food intake versus the decrease in hedonic value during prolonged sensory stimulation.     

Impulse Origin and Propagation in a Bipolar Sensory Neuron

Intracellular recording techniques were used to study electrical activity in bipolar sensory cells associated with crayfish tactile receptors. Several lines of evidence indicate that spikes evoked by natural stimulation of the receptor originate at a dendritic locus. Although overshooting spikes are recorded in the soma in response to both natural and antidromic stimulation receptor potentials are observed only rarely, and, when present, their amplitude is less than 5 mv. Impulses propagating centrifugally into the soma following antidromic stimulation always exhibit an inflection in the rising phase of the spike; however, orthodromic spikes are usually uninflected. Occasionally, orthodromic responses (in the soma) exhibit rather unusual wave forms. Such spikes evoked by natural stimuli are indistinguishable from those elicited electrically in the dendrite, but they do not resemble antidromic impulses. Because the axonal and dendritic boundaries of the soma have a low safety factor for spike transmission, at high frequencies invasion of the soma by dendritic spikes is impeded and often blocked. The soma region can thus act as a low-pass filter. The significance of this self-limiting mechanism for the behavior of the animal is not known; it is suggested, however, that this impediment is a potentially critical one, and may, in other situations, have encouraged the evolution of alternative arrangements.     

Olfactory response and resource utilization in Drosophila: interspecific comparisons

Olfactory response and resource utilization in Drosophila were compared among three domestic (D. melanogaster, D. simulans, D. immigrans) and one Australian endemic (D. lativittata) species. Olfactory response was measured in a choice type olfactometer (Fuyama, 1976). The following chemicals common in Drosophila resources were used as odourants: acetaldehyde, acetic acid, propionic acid, methanol, ethanol, n-propanol, isopropanol, n-butanol. Resource status of these chemicals was determined either from the literature or by adult longevity tests.
All species were attracted by acetaldehyde, while methanol, isopropanol and n-butanol were unattractive. Ethanol attracted all species except D. immigrans , while only D. lativittata and D. melanogaster were attracted to n-propanol, propionic acid and acetic acid
Methanol and isopropanol were not utilized as resources by any of the species, while D. melanogaster and D. lativittata showed greater utilization/tolerance of the other chemicals. Some correlation between resource utilization and olfactory response was found at the interspecific level, although not all chemicals utilized as resources are attractants. The adaptive significance of the interspecific variation in olfactory response is discussed, especially in relation to habitats selected. The results provide suggestions for habitat selection studies at the intraspecific level.     

Impact of Different Carbon Dioxide Concentrations in the Olfactory Response of Sipha flava (Hemiptera: Aphididae) and its Predators

The increasing level CO₂ may altered host plant physiology and hence affect the foraging behavior of herbivore insects and predator. Hence, the aim of this study was provides evidence that host plants grown at different levels of CO₂ can alter the choice behavior of aphid, Sipha flava and their natural enemies, Cycloneda sanguinea and Diomus seminulus. The plant used was Pennisetum purpureum, cultivar Cameron Piracicaba growing in greenhouse (mean value of CO₂?=?440 ppm), climatic chamber with constant value of CO₂?=?500 ppm and climatic chamber with fluctuating CO₂ (mean value?=?368 ppm). A glass Y-shape olfactometer was used to verify the insects responses towards elephant grass plants cultivated under different conditions. The aphids were statistically more attracted by plants grown with constant CO₂ level (500 ppm) than by plants grown with fluctuating CO₂ level or plants grown in greenhouse. There was no difference in S. flava preference to non-infested versus infested plants by conspecifics. The predator C. sanguinea did not show difference between plants grown with constant CO₂ level and infested or not with S. flava. However, the predator D. seminulus showed higher preference to plants grown with constant CO₂ level and infested with S. flava. This study showed that the response of S. flava and its predators were affected by plants grown under different levels of CO₂.