Olfactory Output Research Articles

Engineering Saccharomyces cerevisiae‐based biosensors for copper detection

Heavy metals, that is Cu(II), are harmful to the environment. There is an increasing demand to develop inexpensive detection methods for heavy metals. Here, we developed a yeast biosensor with reduced-noise and improved signal output for potential on-site copper ion detection. The copper-sensing circuit was achieved by employing a secondary genetic layer to control the galactose-inducible (GAL) system in Saccharomyces cerevisiae. The reciprocal control of the Gal4 activator and Gal80 repressor under copper-responsive promoters resulted in a low-noise and sensitive yeast biosensor for copper ion detection. Furthermore, we developed a betaxanthin-based colorimetric assay, as well as 2-phenylethanol and styrene-based olfactory outputs for the copper ion detection. Notably, our engineered yeast sensor confers a narrow range switch-like behaviour, which can give a 'yes/no' response when coupled with a betaxanthin-based visual phenotype. Taken together, we envision that the design principle established here might be applicable to develop other sensing systems for various chemical detections.

Urban spontaneous vegetation helps create unique landsenses

ABSTRACT Non-cultivated, spontaneous vegetation in urban environments plays a key role in supporting urban and rural disturbed ecosystems because of the advantages of low input and low energy consumption. There is a contradiction between such plants and conventional cultivated species in terms of sensory characteristics that could affect people’s acceptance of such vegetated landscapes. Taking the most developed city of China Mainland, Shanghai, as a case study, this work aims to better understand how the public perceive these spontaneous plants in terms of physical and psychological senses based on the recently developed framework of landsenses ecology. Overall, these species activate people with positive senses, such as naturalism and nostalgia. Most people support the importance of their visual aspect, such as colourful fruits/flowers are more important for prompting such species. Our study indirectly supports the idea that the public seem not concern whether plants have soft or hard, smooth or rough tactility in parks. Similarly, people do not make high demands on the plants’ olfactory output. These results may provide a wide range of space for design and management strategies since there are abundant choices. The key influencing factor in our study was ‘childhood living environment’ whereas the commonly related factor ‘level of education’ showed no connection. Based on the framework of landsenses ecology that specifically emphasizes incorporating sensory experience into places, this study may help practitioners connect people’s multi-senses to non-cultivated vegetation and help accommodate a wider range of specific ecological attributes, constraints, priorities and people’s concerns in a more human-centric way.

A unique olfactory bulb microcircuit driven by neurons expressing the precursor to glucagon-like peptide 1

The presence of large numbers of local interneurons in the olfactory bulb has demonstrated an extensive local signaling process, yet the identification and purpose of olfactory microcircuits is poorly explored. Because the discrimination of odors in a complex environment is highly dependent on the tuning of information by local interneurons, we studied for the first time the role of preproglucagon (PPG) neurons in the granule cell layer of the olfactory bulb. Combining electrophysiological recordings and confocal microscopy, we discovered that the PPG neurons are a population of cells expressing the precursor of glucagon-like peptide 1 and are glutamatergic; able to modulate the firing pattern of the mitral cells (M/TCs). Optogenetic activation of PPG neurons resulted in a mixed excitation and inhibition that created a multiphasic response shaping the M/TCs firing pattern. This suggests that PPG neurons could drive neuromodulation of the olfactory output and change the synaptic map regulating olfactory coding.

Paracrine and Synaptic Serotonin Have Opposite Effects on Olfactory Processing in Drosophila Via Distinct Signaling Pathways

Virtually all nervous systems are subject to neuromodulation that alters how signals are processed within and between neurons. Deciphering how modulatory neurons integrate within circuits is complicated because their transmitters can be released either synaptically or in a paracrine fashion to activate a multitude of receptors. Moreover, the functional consequences that different sources of a neuromodulator can have on a neuronal circuit is difficult to establish. We used whole-cell recordings, pharmacology, and genetic manipulations in female Drosophila melanogaster to show that serotonin (5-HT) can reach the antennal lobe (AL), an olfactory neuropil, in a paracrine fashion. The DA1 glomerulus in flies is nearly void of direct serotonergic innervation yet is highly sensitive to 5-HT pharmacology. This sensitivity remains even when the only 5-HT immunoreactive neurons in the AL are genetically ablated, suggesting that 5-HT must enter the AL from outside sources. This paracrine 5-HT establishes a basal tone of the modulator that suppresses olfactory responses and is detected by local interneurons (LN) expressing only the excitatory 5-HT7R. This population of LNs postsynaptically inhibits the output neurons of the AL. Conversely, activation of serotonergic neurons directly innervating the AL boosts olfactory output. This modulation is not sensitive to 5-HT7R manipulation. Our results demonstrate that 5-HT can influence distant circuits and have opposing effects on the same network depending on the source and postsynaptic neurons that it targets.

Sensing by Smell: Nanoparticle-Enzyme Sensors for Rapid and Sensitive Detection of Bacteria with Olfactory Output.

We present here a highly efficient sensor for bacteria that provides an olfactory output, allowing detection without the use of instrumentation and with a modality that does not require visual identification. The sensor platform uses nanoparticles to reversibly complex and inhibits lipase. These complexes are disrupted in the presence of bacteria, restoring enzyme activity and generating scent from odorless pro-fragrance substrate molecules. This system provides rapid (15 min) sensing and very high sensitivity (102 cfu/mL) detection of bacteria using the human sense of smell as an output.

Octopamine modulates activity of neural networks in the honey bee antennal lobe

Neuronal plasticity allows an animal to respond to environmental changes by modulating its response to stimuli. In the honey bee (Apis mellifera), the biogenic amine octopamine plays a crucial role in appetitive odor learning, but little is known about how octopamine affects the brain. We investigated its effect in the antennal lobe, the first olfactory center in the brain, using calcium imaging to record background activity and odor responses before and after octopamine application. We show that octopamine increases background activity in olfactory output neurons, while reducing average calcium levels. Odor responses were modulated both upwards and downwards, with more odor response increases in glomeruli with negative or weak odor responses. Importantly, the octopamine effect was variable across glomeruli, odorants, odorant concentrations and animals, suggesting that the octopaminergic network is shaped by plasticity depending on an individual animal’s history and possibly other factors. Using RNA interference, we show that the octopamine receptor AmOA1 (homolog of the Drosophila OAMB receptor) is involved in the octopamine effect. We propose a network model in which octopamine receptors are plastic in their density and located on a subpopulation of inhibitory neurons in a disinhibitory pathway. This would improve odor-coding of behaviorally relevant, previously experienced odors.Electronic supplementary materialThe online version of this article (doi:10.1007/s00359-013-0805-y) contains supplementary material, which is available to authorized users.

Dye fills reveal additional olfactory tracts in the protocerebrum of wild‐type Drosophila

The antennal lobe (AL) is the primary olfactory center in insect brains. It receives sensory input from the olfactory sensory neurons (OSNs) and sends, through its projection neurons (PNs), reformatted output to secondary olfactory centers, including the mushroom body (MB) calyx and the lateral horn (LH) in the protocerebrum. By injecting dye into the AL of wild-type Drosophila, we identified previously unknown direct pathways between the AL and the ventrolateral, superior medial, and posterior lateral protocerebra. We found that most of these areas in the protocerebrum are connected with the AL through multiple tracts, suggesting that these areas are sites of convergence for olfactory information. Furthermore, areas such as the superior medial protocerebrum now appear to receive olfactory output both directly from the AL and indirectly from lobes of the MB and the LH, suggesting a degree of functional interaction among these areas. We also analyzed the length and number of fibers in each tract. We compare our results obtained from wild-type flies with recent results from transgenic strains and discuss how information about odorants is distributed to multiple protocerebral areas.

Olfactory modulation by dopamine in the context of aversive learning

The need to detect and process sensory cues varies in different behavioral contexts. Plasticity in sensory coding can be achieved by the context-specific release of neuromodulators in restricted brain areas. The context of aversion triggers the release of dopamine in the insect brain, yet the effects of dopamine on sensory coding are unknown. In this study, we characterize the morphology of dopaminergic neurons that innervate each of the antennal lobes (ALs; the first synaptic neuropils of the olfactory system) of the moth Manduca sexta and demonstrate with electrophysiology that dopamine enhances odor-evoked responses of the majority of AL neurons while reducing the responses of a small minority. Because dopamine release in higher brain areas mediates aversive learning we developed a naturalistic, ecologically inspired aversive learning paradigm in which an innately appetitive host plant floral odor is paired with a mimic of the aversive nectar of herbivorized host plants. This pairing resulted in a decrease in feeding behavior that was blocked when dopamine receptor antagonists were injected directly into the ALs. These results suggest that a transient dopaminergic enhancement of sensory output from the AL contributes to the formation of aversive memories. We propose a model of olfactory modulation in which specific contexts trigger the release of different neuromodulators in the AL to increase olfactory output to downstream areas of processing.

Reconstructing the population activity of olfactory output neurons that innervate identifiable processing units

We investigated the functional organization of the moth antennal lobe (AL), the primary olfactory network, by integrating single-cell electrophysiological recording data with geometrical information. The moth AL contains about 60 processing units called glomeruli that are identifiable from one animal to another. We were able to monitor the output information of the AL by recording the activity of a population of output neurons, each of which innervated a single glomerulus. Using compiled in vivo intracellular recordings and staining data from different animals, we mapped the odor-evoked dynamics on a digital atlas of the AL and geometrically reconstructed the population activity. We examined the quantitative relationship between the similarity of olfactory responses and the anatomical distance between glomeruli. Globally, the olfactory response profile was independent of the anatomical distance, although some local features were present. Olfactory response profiles of superficial glomeruli were approximately similar, whereas those of deep glomeruli were different with each other, suggesting network architectures are different in superficial and deep glomerular networks during olfactory processing.

Changes in the connections of the main olfactory bulb after mitral cell selective neurodegeneration

The connections of the main olfactory bulb (OB) of the mouse were studied with iontophoretic injections of biotinylated dextran amine. To sort efferences from mitral cells and tufted cells, the Purkinje cell degeneration (PCD) mouse was used. This mutant animal undergoes a specific neurodegeneration of mitral cells, whereas tufted cells do not degenerate. The unilateral tracer injections used were small and confined largely to the OB of both PCD and control mice at P120. Seven days after tracer injection, the efferences from the OB and the centrifugal afferences from secondary olfactory structures to it were studied. Although there is a large overlap of their target fields, mitral cell axons innervated more caudal regions of the olfactory cortex than tufted cell axons, thus providing definitive evidence of the differential projections of olfactory output neurons. Additionally, an important increase in retrogradely-labeled neurons was detected in the ipsilateral anterior olfactory nucleus of the mutant animals. This was not observed in any other secondary olfactory structure, suggesting a strengthening of the centrifugal input to the OB from that central area after mitral cell loss. Moreover, we recorded a complete loss of bilaterality in the olfactory connections of the PCD mice due to degeneration of the anterior commissure. These results point to an important reorganization of this essential olfactory circuit between the anterior olfactory nucleus and the OB, and hint at a transsynaptic level of plasticity not considered previously in literature.

Air Movement Evokes Electro-Olfactogram Oscillations in the Olfactory Epithelium and Modulates Olfactory Processing in a Slug

In many animals, neurons in the olfactory system have been shown to respond not only to odorants but also to air movements. However, the manner in which the mechanical dynamics of odor stimulation affect olfactory processing remains poorly understood. Using a series of flow rates and odor concentrations from clean air to high-concentration vapors, we systematically analyzed the effects of air movement and odor concentration on olfactory processing. We extracellularly recorded local field potentials and spike units from the olfactory epithelium (OE) and tentacular nerve (TN), which connects the first and second relay centers of olfactory information, in the terrestrial slug Limax marginatus. We found that clean air puffs at a flow rate of 0.18 ml/s (gentle wind), but not high-concentration odor puffs at lower flow rates, induced electro-olfactogram (EOG) oscillations in the OE with a constant frequency (2.5 Hz), regardless of the odor. Surgically isolated OE preparations also showed these EOG oscillations, indicating that the oscillations arose from the OE independently of the downstream circuits. The EOG oscillations entrained the slower spontaneous TN oscillations (1-2 Hz) to the fixed rhythm (2.5 Hz). Spontaneous and odor-evoked units were phase-locked to the TN oscillation peaks. This TN oscillation entrainment by the EOG oscillations caused stronger phase-locking, specifically TN oscillation peaks and EOG oscillation troughs. Taken together, these results suggest that when odors are carried by a gentle wind, the air movement induces EOG oscillations and modulates rhythmic spike patterning of olfactory outputs to the second olfactory relay center in Limax.

Role of inhibition for temporal and spatial odor representation in olfactory output neurons: a calcium imaging study.

The primary olfactory brain center, the antennal lobe (AL) in insects or the olfactory bulb in vertebrates, is a notable example of a neural network for sensory processing. While physiological properties of the input, the olfactory receptor neurons, have become clearer, the operation of the network itself remains cryptic. Therefore we measured spatio-temporal odor-response patterns in the output neurons of the olfactory glomeruli using optical imaging in the honeybee Apis mellifera. We mapped these responses to identified glomeruli, which are the structural and functional units of the AL. Each odor evoked a complex spatio-temporal activity pattern of excited and inhibited glomeruli. These properties were odor- and glomerulus-specific and were conserved across individuals. We compared the spatial pattern of excited glomeruli to previously published signals, which derived mainly from the receptor neurons, and found that they appeared more confined, showing that inhibitory connections enhance the contrast between glomeruli in the AL. To investigate the underlying mechanisms, we applied GABA and the GABA-receptor antagonist picrotoxin (PTX). The results show the presence of two separate inhibitory networks: one is GABAergic and modulates overall AL activity, the other is PTX-insensitive and glomerulus-specific. Inhibitory connections of the latter network selectively inhibit glomeruli with overlapping response profiles, in a way akin to "lateral" inhibition in other sensory systems. Selectively inhibited glomeruli need not be spatial neighbors. The net result is a globally modulated, contrast-enhanced and predictable representation of odors in the olfactory output neurons.