Name: single sensillum recordings for locust palp sensilla basiconica
Uploaded: 2018-06-23T15:00:00.000+00:00
Duration: 7 min 16 s
Description: Watch this Scientific Journal Video about Single Sensillum Recordings for Locust Palp Sensilla Basiconica at JoVE.com

Ce travail est soutenu par une subvention de la Fondation nationale des sciences naturelles de Chine (No.31472037). Toute mention de noms commerciaux ou des produits commerciaux dans cet article est uniquement dans le but de fournir des informations précises et n’implique pas une recommandation.

1. préparation des Instruments et des insectes
<ol><li>
Préparation de tungstène électrodes et des stimuli solutions
	<ol>
<li>Fixer un nouveau fil de tungstène (diamètre de 0,125 mm, longueur 75 mm) dans un micromanipulateur et aiguiser dans une solution de nitrite (NaNO2) sodium 10 % (p/v) dans une seringue à 10 V fourni par une source d’alimentation pendant environ 1 min avec un stéréomicroscope (grossissement de 40 X).</li>
		<li>Tremper le fil de tungstène affûté à plusieurs reprises dans la solution2 NaNO de 10 %, environ 4 mm 5 V < 1 min (Figure 1 a).</li>
		<li>Examiner le diamètre de la pointe de tungstène affûté fréquemment sous le stéréomicroscope jusqu'à ce qu’il fait assez beau pour pénètrent la cuticule d’une sensille olfactive palpe du criquet pèlerin (Figure 1 b).</li>
		<li>Préparer les solutions de stimulation. Diluer chacun de la substance chimique stimulus dans l’huile minérale. Diluer 1-nonanol et acide nonanoïque à des dilutions de 10 %. Diluer le E-2-hexénal et hexanal à 10-2, 10-3, 10-4et 10-5.</li>
		<li>Préparer les tubes transportant les stimuli de Pasteur : insérer des bandes de papier filtre (longueur de 2 cm, largeur de 0,5 cm) dans les tubes de Pasteur, ajouter les solutions de stimulation dilué (chaque 10 µl) pour les bandes de papier filtre et puis branchez les tubes de Pasteur avec pointes de pipette (1 ml).</li>
	</ol>
</li>
	<li>
Préparer l’insecte
	<ol>
<li>Arrière criquets (Locusta migratoria) avec des plantules de blé frais dans des conditions surpeuplées à une humidité relative de 60 %, une température de 28-30 ° C et une photopériode de 18:6 h (lumière : obscurité). Choisissez 1 à 3-jour-âgé de 5ème stade larvaire des larves de Criquet pèlerin et retirer les antennes avec des ciseaux fins pour éviter toute interférence lors de l’enregistrement.</li>
	</ol>
</li>
	<li>
Préparer le titulaire du palpe maxillaire locust
	<ol>
<li>Utiliser une lame de verre (25 mm x 75 mm) comme la base du palpe maxillaire porte (MPH). Attacher un morceau de plastique (1 mm de hauteur, 10 mm de largeur et 35 mm de longueur) à un coin de la lame de verre avec un ruban adhésif double-face et enfin fixer une lamelle couvre-objet (18 x 18 mm) sur le dessus de la pièce en plastique avec adhésif double-face. Placer un petit morceau de ruban de caoutchouc rouge sur la lamelle couvre-objet comme une couche anti-dérapante. La pièce en plastique et le couvercle en verre constituent la plateforme pour le palp locust. La hauteur de la plate-forme est d’environ 1,5 mm.</li>
		<li>Installer un fil de tungstène (de 0,125 mm de diamètre, longueur de 36 mm) à une distance de 1,5 mm parallèle à l’intérieur bord du quai. Fixer les deux extrémités du fil sur la plateforme avec un ruban adhésif double-face.</li>
	</ol>
</li>
</ol>2. préparation des palpes maxillaires Locust
<ol><li>Couper un tube à centrifuger (1,5 ml) verticalement en deux et couper le bas. Placer le pèlerin dans le tube de préparation. Quitter la région ventrale et la tête du pèlerin exposé. Fixer l’ensemble à la lame de verre avec un ruban adhésif double-face (Figure 2 a).</li>
	<li>Tirez le palpe maxillaire droite sur la plate-forme.</li>
	<li>Mettre le fil de tungstène au quatrième segment de la palpe. Placez mastic adhésif de chaque côté du fil de tungstène, environ 2 mm de la palpe maxillaire (Fig. 2 a et 2 b).</li>
</ol>3. seul sensille Recordings
<ol><li>Placer la préparation du palpe maxillaire locust sous un microscope à un faible grossissement (X 100). Ajustez la position de la préparation jusqu'à ce que le palpe est perpendiculaire à l’électrode d’enregistrement (Figure 3 a).</li>
	<li>Insérer l’électrode de référence (électrode de tungstène) dans le œil de Criquet pèlerin à l’aide d’un micromanipulateur. Déplacer l’électrode d’enregistrement (électrode de tungstène) proche du palpe maxillaire avec le micromanipulateur (Figure 3 b et 3C).</li>
	<li>Ajuster le dispositif d’administration odeur à environ 1 cm de la palpe maxillaire (Figure 3 b).</li>
	<li>Ouvrez le logiciel d’enregistrement Auto Spike 32. Définissez les paramètres d’enregistrement comme suit : l’échelle de l’enregistrement sur 500 µV ; le niveau de coupure haute du filtre sur 300 Hz, le faible niveau de coupure sur 200 Hz ; et le pretrigger sur 10 s.</li>
	<li>Connecter l’électrode d’enregistrement pour un 10 x amplificateur AC/DC universel.</li>
	<li>Passer au microscope à un fort grossissement (X 500). Insérez l’électrode d’enregistrement dans la base d’une sensille basiconiques sur le palpe maxillaire et réglez délicatement l’électrode d’enregistrement afin d’obtenir de bonnes pointes spontanées (Figure 3D).</li>
	<li>Ouvrir le contrôleur de stimulation afin d’offrir un flux d’air continu à 20 ml/s. régler l’heure de la stimulation à 1 s. signaux d’enregistrement pour 10 s, à partir de 10 s avant le début de l’impulsion de stimulus.</li>
	<li>Un amplificateur d’AC/DC universel 10 x permet d’amplifier les signaux. Nourrir les signaux dans les 4 IDAC. Analyser les signaux avec le logiciel Auto Spike 32. Des signaux AC sont passe-bande filtré entre 200 à 300 Hz. utilisation Auto Spike 32 pour distinguer les amplitudes de crête à creux de bruits. Calculer les réponses des neurones comme l’augmentation des fréquences de potentiel d’action (pointes par seconde) sur la fréquence spontanée. Effectuer une analyse statistique en utilisant GraphPad Prism 7.</li>
</ol>

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Insectes s’appuient sur les palpes pour détecter les odeurs de nourriture, et leurs palpes sont censées jouer un rôle important dans la spéciation13,27. Les palpes sont de simples organes olfactifs et suscitent un intérêt croissant comme un modèle attrayant pour l’exploration de le neuromoléculaire réseaux sous-jacents chémosensation28.
Insecte labellar et palpe SSRs ont été réalisées avec succ...

Animaux ont développé une gamme d’organes chemosensoriels qui détectent les signaux chimiques exogènes. Chez les insectes, les plus importants organes chemosensoriels sont les antennes et les palpes. Sur ces organes, plusieurs types de poils chemosensoriels, appelés chemosensoriels sensilles, sont innervés par les neurones chemosensoriels (SCSN) dans les poils. Notes de sensilles chemosensoriels reconnaissent les signaux chimiques spécifiques par le biais de transduction d’un signal de stimulus chimiques à des potentiels électriques qui sont ensuite transférées jusqu’au système nerveux central1,,2,3 .
SCSN exprimant divers récepteurs chemosensoriels [p. ex., récepteurs odorants (ORs)], ionotropiques récepteurs (IRs) et les récepteurs gustatifs (GRs) sur leurs membranes, qui codent des signaux chimiques exogènes liés à différents types de chémosensation 4,5,6. La caractérisation des notes de stratégie est essentielle pour l’élucidation des mécanismes moléculaires et cellulaires d’insecte chemoreception. Maintenant seule sensille enregistrement (SSR) est une technique largement utilisée pour la caractérisation d’insecte notes dans les sensilles de nombreux insectes, y compris les mouches7,8de papillons, coléoptères9, pucerons,10,11de criquets, et 12de fourmis. Cependant, peu d’études ont appliqué un SSR à insectes palpes13,14,15,16,17, parce que les structures particulières de leurs sensilles font une enregistrement électrophysiologique difficile18.
Des essaims de criquets pèlerins (Orthoptera) causent souvent des dommages aux cultures grave et une perte économique19. Les palpes sont censées jouer un rôle important dans le choix des aliments des criquets20,21,22,23,24. Deux types de sensilles chemosensoriels sont étudiés par un microscope électronique à balayage (SEM). En général, 350 sensilles chétiformes et 7-8 sensilles basiconiques sont observées sur chaque dôme du criquet pèlerin palpes18. Sensilles chétiformes sont des sensilles gustatives qui détectent les signaux chimiques non volatile, tandis que sensilles basiconiques ont une fonction olfactive, détection de signaux chimiques volatils.
Palpes de Criquet pèlerin, les diamètres des sockets cheveux des sensilles basiconiques (env. 12 µm), sont beaucoup plus élevées que celles des sensilles chétiformes (ca. 8 µm)18,25. La paroi cuticulaire de le basiconica de sensilles sur les palpes est beaucoup plus épaisse que celle des sensilles18. En outre, le dôme de la palpe a fluide contenu dans une cuticule très flexible. Ces caractéristiques font qu’une pénétration avec une microélectrode et une acquisition de bons signaux électrophysiologiques est plus difficile que pour les sensilles. Dans cet article, un protocole SSR détaillé et très efficace pour locust palp sensilles basiconiques est présente avec une vidéo.

<table><tbody><tr><td>Tungsten wire</td><td>ADVENT</td><td>W559504</td><td>Used for making the electrode and fixing the palp</td></tr><tr><td>NaNO&lt;sub&gt;2&lt;/sub&gt;</td><td>Sigma-aldrich</td><td>563218-25G</td><td>Used for sharpening the tungsten wire</td></tr><tr><td>AC Power Supply</td><td>Syntech</td><td>A2-70</td><td>Providing the voltage in sharpening the tungsten wire</td></tr><tr><td>Stereoscope</td><td>Motic</td><td>SMZ-163</td><td>Used for observing the sharpening of tungsten wire</td></tr><tr><td>Microscope</td><td>Olympus</td><td>W-51</td><td>Used for observing the sensilla on locust maxillary palp</td></tr><tr><td>Intelligent Data Acquisition Controller</td><td>Syntech</td><td>IDAC-4</td><td>Real-time on screen display of all signals before and during recording</td></tr><tr><td>Stimulus controller</td><td>Syntech</td><td>CS-55</td><td>Used for controlling the stimulus application</td></tr><tr><td>Electronic micromanipulator</td><td>C.M.D.T</td><td>CFT-8301D</td><td>Used for minor movement of the recording electrode</td></tr><tr><td>Micromanipulator</td><td>Narishige</td><td>MN-151</td><td>Used for minor movement of the reference electrode</td></tr><tr><td>Speaker</td><td>EDIFIER</td><td>R101T06</td><td>Connected with IDAC-4 and providing sound for the signal</td></tr><tr><td>Magnetic base</td><td>PDOK</td><td>PD-101</td><td>Used to hold the electrode, and stimulus delivery tube</td></tr><tr><td>Vibration Isolation Table</td><td>TianHe</td><td>HAP-100-1208</td><td>Used for isolating the vibration from the equipment</td></tr><tr><td>Glass slide</td><td>CITOGLAS</td><td>ZBP-407</td><td>Used for making the base for the MPH</td></tr><tr><td>Blu-tack</td><td>Bostik</td><td>Blu-tack-45g</td><td>Fixing the tungsten wire</td></tr><tr><td>Pasteur tube</td><td>YARE</td><td>WITEG</td><td>Placing the filter paper containing stimuli stimulus solutions</td></tr></tbody></table>

single sensillum recordings for locust palp sensilla basiconica

Les palpes de pièces buccales de Criquet pèlerin sont considérés comme des organes gustatifs classiques qui jouent un rôle important dans le choix des aliments d’un criquet, en particulier pour la détection des signaux chimiques non volatile par l’intermédiaire de sensilles chétiformes (précédemment nommé sensilles terminales (ou sensilles huppés). Il y a maintenant de plus en plus évident que ces palpes ont également une fonction olfactive. Un récepteur odorant (LmigOR2) et une protéine de liaison odorant (LmigOBP1) ont été localisés dans les neurones et les cellules accessoires, respectivement, dans le basiconica de sensilles des palpes. Sensille unique enregistrement (SSR) est utilisé pour enregistrer les réponses des neurones de récepteur odorant, qui est une méthode efficace de dépistage actifs ligands sur les récepteurs spécifiques odorant. SSR est utilisé dans les études fonctionnelles des récepteurs odorants dans des sensilles palpe. La structure de la basiconica de sensilles située sur le dôme des palpes diffère quelque peu de la structure de celles sur les antennes. Donc, lorsque vous effectuez une SSR provoquée par les substances odorantes, quelques conseils spécifiques peut être utile pour l’obtention de résultats optimaux. Dans cet article, un protocole détaillé et très efficace pour un SSR de palp insectes sensilles basiconiques est introduit.

Deux sous-types de sensilles (pb1 et pb2) sur le palpe maxillaire de Criquet pèlerin sont identifiées basées sur la dynamique de réponse différente à des substances chimiques odorantes (10 % 1-nonanol et 10 % d’acide nonanoïque). Les neurones en pb1 produisent significativement plus de pics pour 1-nonanol qu’à l’acide nonanoïque tandis que les neurones en pb2 sont significativement que moins activés par 1-nonanol par rapport à l’acide nonanoïque (F...

Watch this Scientific Journal Video about Single Sensillum Recordings for Locust Palp Sensilla Basiconica at JoVE.com

Single Sensillum Recordings for Locust Palp Sensilla Basiconica

Cet article décrit un protocole détaillé et très efficace pour les enregistrements de sensille unique de la basiconica de sensilles sur les palpes des insectes pièces buccales.

Sensille unique Recordings pour Locust Palp sensilles basiconiques

The palps of locust mouthparts are considered to be conventional gustatory organs that play an important role in a locust&#39;s food selection, especially ...

single-sensillum-recordings-for-locust-palp-sensilla-basiconica

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

8.1K vues.  China Agricultural University. Cet article décrit un protocole détaillé et très efficace pour les enregistrements de sensille unique de la basiconica de sensilles sur les palpes des insectes pièces buccales.

Vidéo: Single Sensillum Recordings for Locust Palp Sensilla Basiconica

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed in sensory hairs called sensilla, which cover the surface of olfactory organs. The surface of each sensillum is covered with tiny pores, through which odorants pass and dissolve in a fluid called sensillum lymph, which bathes the sensory dendrites of the OSNs housed in a given sensillum. The OSN dendrites express odorant receptor (OR) proteins, which in insects function as odor-gated ion channels4, 5. The interaction of odorants with ORs either increases or decreases the basal firing rate of the OSN. This neuronal activity in the form of action potentials embodies the first representation of the quality, intensity, and temporal characteristics of the odorant6, 7.
Given the easy access to these sensory hairs, it is possible to perform extracellular recordings from single OSNs by introducing a recording electrode into the sensillum lymph, while the reference electrode is placed in the lymph of the eye or body of the insect. In Drosophila, sensilla house between one and four OSNs, but each OSN typically displays a characteristic spike amplitude. Spike sorting techniques make it possible to assign spiking responses to individual OSNs. This single sensillum recording (SSR) technique monitors the difference in potential between the sensillum lymph and the reference electrode as electrical spikes that are generated by the receptor activity on OSNs1, 2, 8. Changes in the number of spikes in response to the odorant represent the cellular basis of odor coding in insects. Here, we describe the preparation method currently used in our lab to perform SSR on Drosophila melanogaster and Anopheles gambiae, and show representative traces induced by the odorants in a sensillum-specific manner.

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Electrophysiological responses of olfactory sensory neurons to odorants can be measured in insects using single sensillum recordings. In this video article we will demonstrate how to perform single sensillum recordings in the antennae of the vinegar fly (Drosophila melanogaster) and the maxillary palps of the malaria mosquito (Anopheles gambiae).

Enregistrements Sensillum simple chez les insectes Drosophila melanogaster Et Anopheles gambiae</em

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

Recherche

Neurosciences

Electrophysiological Measurements from a Moth Olfactory System

Insect olfactory systems provide unique opportunities for recording odorant-induced responses in the forms of electroantennograms (EAG) and single sensillum recordings (SSR), which are summed responses from all odorant receptor neurons (ORNs) located on the antenna and from those housed in individual sensilla, respectively. These approaches have been exploited for getting a better understanding of insect chemical communication. The identified stimuli can then be used as either attractants or repellents in management strategies for insect pests.

Insect olfactory systems provide unique opportunities for recording odorant-induced responses in the forms of electroantennograms (EAG) and single sensillum recordings (SSR), which are summed responses from all odorant receptor neurons (ORNs) located on the antenna and from those housed in individual sensilla, respectively.

Mesures électrophysiologiques à partir d&#39;un système de Moth olfactive

Insect olfactory systems provide unique opportunities for recording odorant-induced responses in the forms of electroantennograms (EAG) and single ...

Electrophysiology of Scorpion Peg Sensilla

We describe a modification of an existing tip-recording technique1,2 for electrophysiologically investigating short, peg-like sensory sensilla3,4. On the mid-ventral surface of all scorpions are two appendages called pectines, which have dense fields of mechano- and chemosensory peg sensilla5,6. One method for assessing chemoresponsiveness of these sensilla uses a tungsten electrode for extracellularly recording neural activity within a sensillum as a volatile odorant is introduced to the sensory field5,7. The limitations of this method include slow data collection and uncontrolled stimulant introduction to, and removal from, the peg field. To overcome these limitations, we developed a new tip-recording technique that uses nonpolar mineral oil as a medium through which to deliver water-based tastants to individual peg sensilla8,9. We have successfully applied this method to obtain sensillar chemoresponses to citric acid, ethanol, and salt. Here we describe the experimental protocol for such a study9. We think this new method may be useful for studying the response properties of other arthropod chemosensory systems, including those of insects10, 11 and crustaceans12.

This article describes an electrophysiological method for isolating chemical stimulation to individual sensilla via extracellular, tip-recordings under mineral oil.

Électrophysiologie de Scorpion Peg Sensilles

We describe a modification of an existing tip-recording technique1,2 for electrophysiologically investigating short, peg-like sensory sensilla3,4. On the ...

Sample Preparation Method of Scanning and Transmission Electron Microscope for the Appendages of Woodboring Beetle

This report described sample preparation methods that scanning and transmission electron microscope observations, demonstrated by preparing appendages of the woodboring beetle, Chlorophorus caragana Xie &#38; Wang (2012), for both types of electron microscopy. The scanning electron microscopy (SEM) sample preparation protocol was based on sample chemical fixation, dehydration in a series of ethanol baths, drying, and sputter-coating. By adding Tween 20 (Polyoxyethylene sorbitan laurate) to the fixative and the wash solution, the insect body surface of woodboring beetle was washed more cleanly in SEM. This study&#39;s transmission electron microscopy (TEM) sample preparation involved a series of steps including fixation, ethanol dehydration, embedding in resin, positioning using fluorescence microscopy, sectioning, and staining. Fixative with Tween 20 enabled penetrate the insect body wall of woodboring beetle more easily than it would had been without Tween 20, and subsequently better fixed tissues and organs in the body, thus yielded clear transmission electron microscope observations of insect sensilla ultrastructures. The next step of this preparation was determining the positions of insect sensilla in the sample embedded in the resin block by using fluorescence microscopy to increase the precision of target sensilla positioning. This improved slicing accuracy.

In order to observe ultrastructure of insect sensilla, scanning and transmission electron microscopy (SEM and TEM, respectively) sample preparation protocol were presented in the study. Tween 20 was added into the fixative to avoid sample deformation in SEM. Fluorescence microscopy was helpful for improving slicing accuracy in TEM.

Méthode de préparation d'échantillons de microscope électronique de balayage et de transmission pour les appendices du bec de bois

This report described sample preparation methods that scanning and transmission electron microscope observations, demonstrated by preparing appendages of ...

Environnement

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

Méthodes d&#39;enregistrement multi-unités pour caractériser l&#39;activité neuronale dans le Criquet (<em&gt; Schistocerca americana</em&gt; Circuits olfactifs)

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Electrophysiological Recording From Drosophila Labellar Taste Sensilla

The peripheral taste response of insects can be powerfully investigated with electrophysiological techniques. The method described here allows the researcher to measure gustatory responses directly and quantitatively, reflecting the sensory input that the insect nervous system receives from taste stimuli in its environment. This protocol outlines all key steps in performing this technique. The critical steps in assembling an electrophysiology rig, such as selection of necessary equipment and a suitable environment for recording, are delineated. We also describe how to prepare for recording by making appropriate reference and recording electrodes, and tastant solutions. We describe in detail the method used for preparing the insect by insertion of a glass reference electrode into the fly in order to immobilize the proboscis. We show traces of the electrical impulses fired by taste neurons in response to a sugar and a bitter compound. Aspects of the protocol are technically challenging and we include an extensive description of some common technical challenges that may be encountered, such as lack of signal or excessive noise in the system, and potential solutions. The technique has limitations, such as the inability to deliver temporally complex stimuli, observe background firing immediately prior to stimulus delivery, or use water-insoluble taste compounds conveniently. Despite these limitations, this technique (including minor variations referenced in the protocol) is a standard, broadly accepted procedure for recording Drosophila neuronal responses to taste compounds.

Electrophysiological Recording From Drosophila Labellar Taste Sensilla

This protocol describes extracellular recording of the action potential responses fired by labellar taste neurons in Drosophila.

Enregistrement électrophysiologique De<em&gt; Drosophila</em&gt; Labellar Goût Sensilles

The peripheral taste response of insects can be powerfully investigated with electrophysiological techniques. The method described here allows the ...

Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals

The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house single or multiple neurons inside, are considered to make the major contribution in responding to the chemical stimuli. By directly recording action potential in the olfactory sensillum after exposure to stimuli, single sensillum recording (SSR) technique provides a powerful approach for investigating the neural responses of insects to chemical stimuli. For the bed bug, which is a notorious human parasite, multiple types of olfactory sensillum have been characterized. In this study, we demonstrated neural responses of bed bug olfactory sensilla to two chemical stimuli and the dose-dependent responses to one of them using the SSR method. This approach enables researchers to conduct early screening for individual chemical stimuli on the bed bug olfactory sensilla, which would provide valuable information for the development of new bed bug attractants or repellents and benefits the bed bug control efforts.

Bed bugs rely on olfactory receptor neurons housed in their antennal olfactory sensilla to detect semiochemicals in the environment. Utilizing single sensillum recording, we demonstrate a method to evaluate bed bug response to semiochemicals and explore the coding process involved.

Utilisation Sensillum simple enregistrement pour détecter les réponses olfactive Neuron des punaises de lit à écomones

The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house ...

Localization of Odorant Receptor Genes in Locust Antennae by RNA In Situ Hybridization

Insects have evolved sophisticated olfactory reception systems to sense exogenous chemical signals. These chemical signals are transduced by Olfactory Receptor Neurons (ORNs) housed in hair-like structures, called chemosensilla, of the antennae. On the ORNs&#39; membranes, Odorant Receptors (ORs) are believed to be involved in odor coding. Thus, being able to identify genes localized to the ORNs is necessary to recognize OR genes, and provides a fundamental basis for further functional in situ studies. The RNA expression levels of specific ORs in insect antennae are very low, and preserving insect tissue for histology is challenging. Thus, it is difficult to localize an OR to a specific type of sensilla using RNA in situ hybridization. In this paper, a detailed and highly effective RNA in situ hybridization protocol particularly for lowly expressed OR genes of insects, is introduced. In addition, a specific OR gene was identified by conducting double-color fluorescent in situ hybridization experiments using a co-expressing receptor gene, Orco, as a marker.

Localization of Odorant Receptor Genes in Locust Antennae by RNA In Situ Hybridization

This paper describes a detailed and highly effective RNA in situ hybridization protocol particularly for low-level expressed Odorant Receptor (OR) genes, as well as other genes, in insect antennae using digoxigenin (DIG)-labeled or biotin-labeled probes.

Localisation des gènes des récepteurs odorants dans les antennes acridiennes par ARN<em&gt; In Situ</em&gt; Hybridation

Insects have evolved sophisticated olfactory reception systems to sense exogenous chemical signals. These chemical signals are transduced by Olfactory ...

Electrophysiological Recording from Drosophila Trichoid Sensilla in Response to Odorants of Low Volatility

Insects rely on their sense of smell to guide a wide range of behaviors that are critical for their survival, such as food-seeking, predator avoidance, oviposition, and mating. Myriad chemicals of varying volatilities have been identified as natural odorants that activate insect Olfactory Receptor Neurons (ORNs). However, studying the olfactory responses to low-volatility odorants has been hampered by an inability to effectively present such stimuli using conventional odor-delivery methods. Here, we describe a procedure that permits the effective presentation of low-volatility odorants for in vivo Single-Sensillum Recording (SSR). By minimizing the distance between the odor source and the target tissue, this method allows for the application of biologically salient but hitherto inaccessible odorants, including palmitoleic acid, a stimulatory pheromone with a demonstrated effect on ORNs involved in courtship and mating behavior1. Our procedure thus affords a new avenue to assay a host of low-volatility odorants for the study of insect olfaction and pheromone communication.

Electrophysiological Recording from Drosophila Trichoid Sensilla in Response to Odorants of Low Volatility

The overall goal of this protocol is to demonstrate how to present odorants of low volatility for single-sensillum recording from Drosophila olfactory receptor neurons that respond to long-chain cuticular pheromones.

Enregistrement électrophysiologique de<em&gt; Drosophila</em&gt; Sensilla Trichoid en Réponse aux Odorants de Faible Volatilité

Insects rely on their sense of smell to guide a wide range of behaviors that are critical for their survival, such as food-seeking, predator avoidance, ...

Une nouvelle méthode pour une analyse améliorée de l’activité des neurones gustatifs des insectes

Base Recording: A Technique for Analyzing Responses of Taste Neurons in Drosophila

Insects taste the external world through taste hairs, or sensilla, that have pores at their tips. When a sensillum comes into contact with a potential food source, compounds from the food source enter through the pore and activate neurons within. For over 50 years, these responses have been recorded using a technique called tip recording. However, this method has major limitations, including the inability to measure neural activity before or after stimulus contact and the requirement for tastants to be soluble in aqueous solutions. We describe here a technique that we call base recording, which overcomes these limitations. Base recording allows the measurement of taste neuron activity before, during, and after the stimulus. Thus, it allows extensive analysis of OFF responses that occur after a taste stimulus. It can be used to study hydrophobic compounds such as long-chain pheromones that have very low solubility in water. In summary, base recording offers the advantages of single-sensillum electrophysiology as a means of measuring neuronal activity - high spatial and temporal resolution, without the need for genetic tools - and overcomes key limitations of the traditional tip recording technique.

Pleins feux sur l’auteur : Progrès en chimioréception – des récepteurs d’odeurs d’insectes aux ARN non codants

A rarely used method of electrophysiological recording, base recording, allows analysis of features of taste coding that cannot be examined by conventional recording methods. Base recording also allows the analysis of taste responses to hydrophobic stimuli that cannot be studied using traditional electrophysiological methods.

Enregistrement de base : une technique d’analyse des réponses des neurones gustatifs chez la drosophile

Insects taste the external world through taste hairs, or sensilla, that have pores at their tips. When a sensillum comes into contact with a potential ...

Sensille unique Recordings pour Locust Palp sensilles basiconiques

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Chapitres dans cette vidéo

Enregistrements Sensillum simple chez les insectes Drosophila melanogaster Et Anopheles gambiae

Mesures électrophysiologiques à partir d'un système de Moth olfactive

Électrophysiologie de Scorpion Peg Sensilles

Méthode de préparation d'échantillons de microscope électronique de balayage et de transmission pour les appendices du bec de bois

Méthodes d'enregistrement multi-unités pour caractériser l'activité neuronale dans le Criquet (

Enregistrement électrophysiologique De

Utilisation Sensillum simple enregistrement pour détecter les réponses olfactive Neuron des punaises de lit à écomones

Localisation des gènes des récepteurs odorants dans les antennes acridiennes par ARN

Enregistrement électrophysiologique de

Enregistrement de base : une technique d’analyse des réponses des neurones gustatifs chez la drosophile