Name: Single Sensillum Recordings for Locust Palp Sensilla Basiconica (Video) | JoVE Journal
Uploaded: 2018-06-23T15:00:00.000+00:00
Duration: 7 min 16 s
Description: Learn single sensillum recordings from locust palp sensilla basiconica for studying olfactory neuron responses to chemical stimuli. This electrophysiology method reveals dose-dependent firing patterns in insect chemoreception.

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5 minutes

This work is supported by a grant from the National Natural Science Foundation of China (No.31472037). Any mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply a recommendation.

1. Preparation of Instruments and Insect
<ol>
	<li>Preparing tungsten electrodes and stimuli solutions
	<ol>
		<li>Fix a new tungsten wire (diameter of 0.125 mm, length of 75 mm) into a micromanipulator and sharpen it in a 10% (w/v) sodium nitrite (NaNO2) solution in a syringe at 10 V provided by a power supply for about 1 min under a stereomicroscope (40X magnification).</li>
		<li>Dip the sharpened tungsten wire repeatedly into the 10% NaNO2 solution, about 4 mm at 5 V in &#60; 1 min (Figure 1A).</li>
		<li>Examine the diameter of the sharpened tungsten tip frequently under the stereomicroscope until it is fine enough to penetrate the cuticle of a locust palp olfactory sensillum (Figure 1B).</li>
		<li>Prepare the stimulus solutions. Dilute each of the chemical stimulus substance in mineral oil. Dilute 1-nonanol and nonanoic acid at 10% dilutions. Dilute E-2-hexenal and hexanal at 10-2, 10-3, 10-4, and 10-5.</li>
		<li>Prepare Pasteur tubes carrying the stimuli: insert filter paper strips (length of 2 cm, width of 0.5 cm) into the Pasteur tubes, add the diluted stimulus solutions (each 10 &#181;l) to the filter paper strips, and then plug the Pasteur tubes with pipette tips (1 ml).</li>
	</ol>
	</li>
	<li>Prepare the insect
	<ol>
		<li>Rear locusts (Locusta migratoria) with fresh wheat seedlings under crowded conditions at a relative humidity of 60%, a temperature of 28 - 30 &#176;C, and a photoperiod of 18:6 h (light:dark). Choose 1- to 3-day-old 5th instar locust nymphs and remove the antennae with fine scissors to avoid any interference when recording.</li>
	</ol>
	</li>
	<li>Preparing the locust maxillary palp holder
	<ol>
		<li>Use a glass slide (25 mm x 75 mm) as the base of the maxillary palp holder (MPH). Attach a plastic piece (1 mm in height, 10 mm in width, 35 mm in length) to a corner of the glass slide with double-sided adhesive tape, and finally fix a cover glass (18 mm x 18 mm) on top of the plastic piece with double-sided adhesive tape. Place a small piece of red rubber tape onto the cover glass as a non-slip layer. The plastic piece and the cover glass constitute the platform for the locust palp. The height of the platform is approximately 1.5 mm.</li>
		<li>Install a tungsten wire (diameter of 0.125 mm, length of 36 mm) at a distance of 1.5 mm parallel to the inside edge of the platform. Fix the two ends of the wire onto the platform with double-sided adhesive tape.</li>
	</ol>
	</li>
</ol>
2. Preparation of Locust Maxillary Palps
<ol>
	<li>Cut a centrifuge tube (1.5 ml) vertically in half and cut off the bottom. Place the locust into the prepared tube. Leave the ventral region and the head of the locust exposed. Fix the assembly to the glass slide with double-sided adhesive tape (Figure 2A).</li>
	<li>Pull the right maxillary palp onto the platform.</li>
	<li>Put the tungsten wire at the fourth segment of the palp. Place adhesive putty on each side of the tungsten wire, about 2 mm from the maxillary palp (Figure 2A and 2B).</li>
</ol>
3. Single Sensillum Recordings
<ol>
	<li>Place the locust maxillary palp preparation under a microscope at a low magnification (100X). Adjust the position of the preparation until the palp is perpendicular to the recording electrode (Figure 3A).</li>
	<li>Insert the reference electrode (tungsten electrode) into the locust eye using a micromanipulator. Move the recording electrode (tungsten electrode) close to the maxillary palp with the micromanipulator (Figure 3B and 3C).</li>
	<li>Adjust the odor delivery device to about 1 cm from the maxillary palp (Figure 3B).</li>
	<li>Open the recording software Auto Spike 32. Set the recording parameters as follows: the recording scale on 500 &#181;V; the high cutoff of the filter on 300 Hz, the low cutoff on 200 Hz; and the pretrigger on 10 s.</li>
	<li>Connect the recording electrode to a 10x universal AC/DC amplifier.</li>
	<li>Switch the microscope to a high magnification (500X). Insert the recording electrode into the base of a basiconic sensillum on the maxillary palp and delicately adjust the recording electrode to obtain good spontaneous spikes (Figure 3D).</li>
	<li>Open the stimulus controller to deliver a continuous air stream at 20 ml/s. Set the stimulation time to 1 s. Record signals for 10 s, starting 10 s before the onset of the stimulus pulse.</li>
	<li>Use a 10x universal AC/DC amplifier to amplify the signals. Feed the signals into the IDAC 4. Analyze the signals with the Auto Spike 32 software. AC signals are band-pass filtered between 200 to 300 Hz. Use Auto Spike 32 to distinguish peak-to-trough amplitudes from noises. Calculate the responses of the neurons as the increases in action potential frequencies (spikes per second) over the spontaneous frequencies. Perform a statistical analysis using GraphPad Prism 7.</li>
</ol>

<ol><li>Carey, A. F., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Insect+olfaction+from+model+systems+to+disease+control%5BTitle%5D)+AND+%22Proceedings+of+the+National+Academy+of+Sciences+of+the+United+States+of+America%22%5BJournal%5D)">Insect olfaction from model systems to disease control.</a> Proceedings of the National Academy of Sciences of the United States of America. 108 (32), 12987-12995 (2011).</li>
<li>Leal, W. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Odorant+reception+in+insects%3A+roles+of+receptors%2C+binding+proteins%2C+and+degrading+enzymes%5BTitle%5D)+AND+%22Annual+Review+of+Entomology%22%5BJournal%5D)">Odorant reception in insects: roles of receptors, binding proteins, and degrading enzymes.</a> Annual Review of Entomology. 58, 373-391 (2013).</li>
<li>Zhang, J., Walker, W. B., Wang, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pheromone+reception+in+moths%3A+from+molecules+to+behaviors%5BTitle%5D)+AND+%22Progress+in+Molecular+Biology+and+Translational+Science%22%5BJournal%5D)">Pheromone reception in moths: from molecules to behaviors.</a> Progress in Molecular Biology and Translational Science. 130, 109-128 (2015).</li>
<li>Vosshall, L. B., Amrein, H., Morozov, P. S., Rzhetsky, A., Axel, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+spatial+map+of+olfactory+receptor+expression+in+the+Drosophila+antenna%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">A spatial map of olfactory receptor expression in the Drosophila antenna.</a> Cell. 96 (5), 725-736 (1999).</li>
<li>Benton, R., Vannice, K. S., Gomez-Diaz, C., Vosshall, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Variant+ionotropic+glutamate+receptors+as+chemosensory+receptors+in+Drosophila%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">Variant ionotropic glutamate receptors as chemosensory receptors in Drosophila.</a> Cell. 136 (1), 149-162 (2009).</li>
<li>Vosshall, L. B., Stocker, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Molecular+architecture+of+smell+and+taste+in+Drosophila%5BTitle%5D)+AND+%22Annual+Review+of+Neuroscience%22%5BJournal%5D)">Molecular architecture of smell and taste in Drosophila.</a> Annual Review of Neuroscience. 30, 505-533 (2007).</li>
<li>de Bruyne, M., Foster, K., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Odor+coding+in+the+Drosophila+antenna%5BTitle%5D)+AND+%22Neuron%22%5BJournal%5D)">Odor coding in the Drosophila antenna.</a> Neuron. 30, 537-552 (2001).</li>
<li>Roelofs, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Sex+pheromone+production+and+perception+in+European+corn+borer+moths+is+determined+by+both+autosomal+and+sex-linked+genes%5BTitle%5D)+AND+%22Proceedings+of+the+National+Academy+of+Sciences+of+the+United+States+of+America%22%5BJournal%5D)">Sex pheromone production and perception in European corn borer moths is determined by both autosomal and sex-linked genes.</a> Proceedings of the National Academy of Sciences of the United States of America. 84 (21), 7585-7589 (1987).</li>
<li>Larsson, M. C., Leal, W. S., Hansson, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Olfactory+receptor+neurons+detecting+plant+odours+and+male+volatiles+in+Anomala+cuprea+beetles+%28Coleoptera%3A+Scarabaeidae%29%5BTitle%5D)+AND+%22Journal+of+Insect+Physiology%22%5BJournal%5D)">Olfactory receptor neurons detecting plant odours and male volatiles in Anomala cuprea beetles (Coleoptera: Scarabaeidae).</a> Journal of Insect Physiology. 47 (9), 1065-1076 (2001).</li>
<li>Zhang, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Molecular+basis+of+alarm+pheromone+detection+in+aphids%5BTitle%5D)+AND+%22Current+Biology%22%5BJournal%5D)">Molecular basis of alarm pheromone detection in aphids.</a> Current Biology. 27 (1), 55-61 (2017).</li>
<li>Cui, X., Wu, C., Zhang, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electrophysiological+response+patterns+of+16+olfactory+neurons+from+the+trichoid+sensilla+to+odorant+from+fecal+volatiles+in+the+locust%2C+Locusta+migratoria+manilensis%5BTitle%5D)+AND+%22Archives+of+Insect+Biochemistry+and+Physiology%22%5BJournal%5D)">Electrophysiological response patterns of 16 olfactory neurons from the trichoid sensilla to odorant from fecal volatiles in the locust, Locusta migratoria manilensis.</a> Archives of Insect Biochemistry and Physiology. 77 (2), 45-57 (2011).</li>
<li>Sharma, K. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cuticular+hydrocarbon+pheromones+for+social+behavior+and+their+coding+in+the+ant+antenna%5BTitle%5D)+AND+%22Cell+Reports%22%5BJournal%5D)">Cuticular hydrocarbon pheromones for social behavior and their coding in the ant antenna.</a> Cell Reports. 12 (8), 1261-1271 (2015).</li>
<li>de Bruyne, M., Clyne, P. J., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Odor+coding+in+a+model+olfactory+organ%3A+the+Drosophila+maxillary+palp%5BTitle%5D)+AND+%22Journal+of+Neuroscience%22%5BJournal%5D)">Odor coding in a model olfactory organ: the Drosophila maxillary palp.</a> Journal of Neuroscience. 19 (11), 4520-4532 (1999).</li>
<li>Syed, Z., Leal, W. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Maxillary+palps+are+broad+spectrum+odorant+detectors+in+Culex+quinquefasciatus%5BTitle%5D)+AND+%22Chemical+Senses%22%5BJournal%5D)">Maxillary palps are broad spectrum odorant detectors in Culex quinquefasciatus.</a> Chemical Senses. 32 (8), 727-738 (2007).</li>
<li>Lu, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Odor+coding+in+the+maxillary+palp+of+the+malaria+vector+mosquito+Anopheles+gambiae%5BTitle%5D)+AND+%22Current+Biology%22%5BJournal%5D)">Odor coding in the maxillary palp of the malaria vector mosquito Anopheles gambiae.</a> Current Biology. 17 (18), 1533-1544 (2007).</li>
<li>Pellegrino, M., Nakagawa, T., Vosshall, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Single+sensillum+recordings+in+the+insects+Drosophila+melanogaster+and+Anopheles+gambiae%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Single sensillum recordings in the insects Drosophila melanogaster and Anopheles gambiae.</a> Journal of Visualized Experiments. 36, e1725(2010).</li>
<li>Grant, A. J., Wigton, B. E., Aghajanian, J. G., O'Connell, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electrophysiological+responses+of+receptor+neurons+in+mosquito+maxillary+palp+sensilla+to+carbon+dioxide%5BTitle%5D)+AND+%22Journal+of+Comparative+Physiology+A%22%5BJournal%5D)">Electrophysiological responses of receptor neurons in mosquito maxillary palp sensilla to carbon dioxide.</a> Journal of Comparative Physiology A. 177 (4), 389-396 (1995).</li>
<li>Blaney, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+ultrastructure+of+an+olfactory+sensillum+on+the+maxillary+palps+of+Locusta+migratoria+%28L.%29%5BTitle%5D)+AND+%22Cell+and+Tissue+Research%22%5BJournal%5D)">The ultrastructure of an olfactory sensillum on the maxillary palps of Locusta migratoria (L.).</a> Cell and Tissue Research. 184 (3), 397-409 (1977).</li>
<li>Hassanali, A., Njagi, P. G. N., Bashir, M. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Chemical+ecology+of+locusts+and+related+acridids%5BTitle%5D)+AND+%22Annual+Review+of+Entomology%22%5BJournal%5D)">Chemical ecology of locusts and related acridids.</a> Annual Review of Entomology. 50, 223-245 (2005).</li>
<li>Chapman, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Contact+chemoreception+in+feeding+by+phytophagous+insects%5BTitle%5D)+AND+%22Annual+Review+of+Entomology%22%5BJournal%5D)">Contact chemoreception in feeding by phytophagous insects.</a> Annual Review of Entomology. 48, 455-484 (2003).</li>
<li>Chapman, R. F., Sword, G. <a target="_blank" href="https://link.springer.com/article/10.1007/BF01049149">The importance of palpation in food selection by a polyphagous grasshopper (Orthoptera: Acrididae).</a> Journal of Insect Behavior. 6, 79-91 (1993).</li>
<li>Winstanley, C., Blaney, W. <a target="_blank" href="http://onlinelibrary.wiley.com/doi/10.1111/j.1570-7458.1978.tb02839.x/abstract">Chemosensory mechanisms of locusts in relation to feeding.</a> Entomologia Experimentalis et Applicata. 24, 750-758 (1978).</li>
<li>Blaney, W., Duckett, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+significance+of+palpation+by+the+maxillary+palps+of+Locusta+migratoria+%28L.%29%3A+an+electrophysiological+and+behavioural+study%5BTitle%5D)+AND+%22Journal+of+Experimental+Biology%22%5BJournal%5D)">The significance of palpation by the maxillary palps of Locusta migratoria (L.): an electrophysiological and behavioural study.</a> Journal of Experimental Biology. 63, 701-712 (1975).</li>
<li>Blaney, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electrophysiological+responses+of+the+terminal+sensilla+on+the+maxillary+palps+of+Locusta+migratoria+%28L.%29+to+some+electrolytes+and+non-electrolytes%5BTitle%5D)+AND+%22Journal+of+Experimental+Biology%22%5BJournal%5D)">Electrophysiological responses of the terminal sensilla on the maxillary palps of Locusta migratoria (L.) to some electrolytes and non-electrolytes.</a> Journal of Experimental Biology. 60, 275-293 (1974).</li>
<li>Jin, X., Zhang, S., Zhang, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Expression+of+odorant-binding+and+chemosensory+proteins+and+spatial+map+of+chemosensilla+on+labial+palps+of+Locusta+migratoria+%28Orthoptera%3A+Acrididae%29%5BTitle%5D)+AND+%22Anthropod+Structure+%26+Development%22%5BJournal%5D)">Expression of odorant-binding and chemosensory proteins and spatial map of chemosensilla on labial palps of Locusta migratoria (Orthoptera: Acrididae).</a> Anthropod Structure & Development. 35 (1), 47-56 (2006).</li>
<li>Zhang, L., Li, H., Zhang, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Two+olfactory+pathways+to+detect+aldehydes+on+locust+mouthpart%5BTitle%5D)+AND+%22International+Journal+of+Biological+Sciences%22%5BJournal%5D)">Two olfactory pathways to detect aldehydes on locust mouthpart.</a> International Journal of Biological Sciences. 13 (6), 759-771 (2017).</li>
<li>Dweck, H. K. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Olfactory+channels+associated+with+the+Drosophila+maxillary+palp+mediate+short-+and+long-range+attraction%5BTitle%5D)+AND+%22eLife%22%5BJournal%5D)">Olfactory channels associated with the Drosophila maxillary palp mediate short- and long-range attraction.</a> eLife. 5, e14925(2016).</li>
<li>Bohbot, J. D., Sparks, J. T., Dickens, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+maxillary+palp+of+Aedes+aegypti%2C+a+model+of+multisensory+Integration%5BTitle%5D)+AND+%22Insect+Biochemistry+and+Molecular+Biology%22%5BJournal%5D)">The maxillary palp of Aedes aegypti, a model of multisensory Integration.</a> Insect Biochemistry and Molecular Biology. 48, 29-39 (2014).</li>
<li>Delventhal, R., Kiely, A., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electrophysiological+recording+from+Drosophila+labellar+taste+sensilla%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Electrophysiological recording from Drosophila labellar taste sensilla.</a> Journal of Visualized Experiments. 84, e51355(2014).</li>
<li>Ng, R., Lin, H. H., Wang, J. W., Su, C. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electrophysiological+recording+from+Drosophila+trichoid+sensilla+in+response+to+odorants+of+low+volatility%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Electrophysiological recording from Drosophila trichoid sensilla in response to odorants of low volatility.</a> Journal of Visualized Experiments. 125, e56147(2017).</li>
<li>Syed, Z., Leal, W. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electrophysiological+measurements+from+a+moth+olfactory+system%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Electrophysiological measurements from a moth olfactory system.</a> Journal of Visualized Experiments. 49, e2489(2011).</li>
<li>Saha, D., Leong, K., Katta, N., Raman, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Multi-unit+recording+methods+to+characterize+neural+activity+in+the+locust+%28Schistocerca+Americana%29+olfactory+circuits%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Multi-unit recording methods to characterize neural activity in the locust (Schistocerca Americana) olfactory circuits.</a> Journal of Visualized Experiments. 71, e50139(2013).</li>
<li>Liu, F., Liu, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Using+single+sensillum+recording+to+detect+olfactory+neuron+responses+of+bed+bugs+to+semiochemicals%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Using single sensillum recording to detect olfactory neuron responses of bed bugs to semiochemicals.</a> Journal of Visualized Experiments. 107, e53337(2016).</li>
</ol>

The authors have nothing to disclose.

Insects rely on palps to detect food odors, and their palps are believed to play an important role in speciation13,27. The palps are simple olfactory organs and are receiving increasing attention as an attractive model for the exploration of the neuromolecular networks underlying chemosensation28.
Insect labellar and palp SSRs have been successfully performed on Drosophila melanogaster, Anopheles ga...

Animals have evolved a range of chemosensory organs that sense exogenous chemical cues. In insects, the most important chemosensory organs are the antennae and the palps. On these organs, several types of chemosensory hairs, called chemosensory sensilla, are innervated by chemosensory neurons (CSNs) within the hairs. CSNs in chemosensory sensilla recognize specific chemical cues through signal transduction from chemical stimuli to electrical potentials that are subsequently transferred up to the central nervous systems1,2,3.
CSNs express various chemosensory receptors [e.g., odorant receptors (ORs)], ionotropic receptors (IRs), and gustatory receptors (GRs) on their membranes, which encode exogenous chemical cues associated with different types of chemosensation4,5,6. The characterization of CSNs is key to the elucidation of cellular and molecular mechanisms of insect chemoreception. Now single sensillum recording (SSR) is a widely-used technique for the characterization of insect CSNs in the antennal sensilla of many insects, including flies7, moths8, beetles9, aphids10, locusts11, and ants12. However, few studies have applied an SSR to insect palps13,14,15,16,17, because the particular structures of their sensilla make an electrophysiological recording difficult18.
Swarms of locusts (Orthoptera) often cause serious crop damage and economic loss19. The palps are believed to play an important role in the food selection of locusts20,21,22,23,24. Two types of chemosensory sensilla are investigated by a scanning electron microscope (SEM). Usually, 350 sensilla chaetica and 7 - 8 sensilla basiconica are observed on each dome of the locust palps18. Sensilla chaetica are gustatory sensilla that sense non-volatile chemical cues, whereas sensilla basiconica have an olfactory function, sensing volatile chemical cues.
On locust palps, the diameters of the hair sockets of the sensilla basiconica (ca. 12 &#181;m), are much greater than those of sensilla chaetica (ca. 8 &#181;m)18,25. The cuticular wall of the sensilla basiconica on the palps is much thicker than that of antennal sensilla18. In addition, the dome of the palp has fluid contents within a highly flexible cuticle. These characteristics mean that a penetration with a microelectrode and an acquisition of good electrophysiological signals is more difficult than for antennal sensilla. In this paper, a detailed and highly effective SSR protocol for locust palp sensilla basiconica is presented with a video.

<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

Single Sensillum Recordings for Locust Palp Sensilla Basiconica

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 for the detection of non-volatile chemical cues through sensilla chaetica (previously named terminal sensilla or crested sensilla). There is now increasing evidence that these palps also have an olfactory function. An odorant receptor (LmigOR2) and an odorant-binding protein (LmigOBP1) have been localized in the neurons and accessory cells, respectively, in the sensilla basiconica of the palps. Single sensillum recording (SSR) is used for recording the responses of odorant receptor neurons, which is an effective method for screening active ligands on specific odorant receptors. SSR is used in functional studies of odorant receptors in palp sensilla. The structure of the sensilla basiconica located on the dome of the palps differs somewhat from the structure of those on the antennae. Therefore, when performing an SSR elicited by odorants, some specific advice may be helpful for obtaining optimum results. In this paper, a detailed and highly effective protocol for an SSR from insect palp sensilla basiconica is introduced.

Two sensilla subtypes (pb1 and pb2) on the locust maxillary palp are identified based on different response dynamics to chemical odorants (10% 1-nonanol and 10% nonanoic acid). The neurons in pb1 produce significantly more spikes to 1-nonanol than to nonanoic acid while the neurons in pb2 are significantly less activated by 1-nonanol compared with nonanoic acid (Figure 4). Hexanal and E-2-Hexenal can evoke a locust palp opening response (POR)<sup class="xref"...

This paper describes a detailed and highly effective protocol for single sensillum recordings from the sensilla basiconica on the palps of insect mouthparts.

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

Research

JoVE Journal

Neuroscience

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).

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

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.

Electrophysiological Measurements from a Moth Olfactory System

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

Electrophysiology of Scorpion Peg Sensilla

In this video, a live Drosophila fly is mounted in a tube for electrophysiological studies. Electrodes are positioned at the base of the sensillum to record specific neuronal responses to pheromones with the fly's olfactory receptor neurons.

Electrophysiological Recordings from Drosophila Sensilla in Response to Volatile Odorants

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.

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

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

Electrophysiological Recording From Drosophila Labellar Taste Sensilla

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.

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

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.

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

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.

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

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.

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

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.

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

Learn single sensillum recordings from locust palp sensilla basiconica for studying olfactory neuron responses to chemical stimuli. This electrophysiology method reveals dose-dependent firing patterns in insect chemoreception.

Single Sensillum Recordings for Locust Palp Sensilla Basiconica

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