Name: a simple way to measure alterations in reward-seeking behavior using drosophila melanogaster
Uploaded: 2016-12-15T13:00:00
Description: Watch this Scientific Journal Video about A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster at JoVE.com

Ringraziamo U. Heberlein e A. Devineni per le discussioni di lunga durata e consulenza tecnica. Ringraziamo anche i membri del laboratorio Shohat-Ophir, A. Benzur, L. Kazaz, e O. Shalom, per l&#39;aiuto di dimostrare il metodo. apprezzamento speciale va a Eliezer Costi per stabilire i sistemi di mosca in laboratorio. Questo lavoro è stato sostenuto dalla Israel Science Foundation (384/14) e le Marie Curie Career Integration Grants (CIG 631.127).

Nota: panoramica generale del disegno sperimentale: Il disegno sperimentale prevede un protocollo adattato per la soppressione corteggiamento 7-9 in cui le mosche di sesso maschile sono esposti a esperienze gratificanti e nonrewarding in 3 allenamenti consecutivi nel corso di 4 d. Al termine della fase di esperienza, le mosche sono testati in un saggio di consumo di etanolo volontaria due scelta per 3 - 4 d. Il protocollo include qui diverse fasi preparatorie, alcune delle quali possono essere fatte in anticipo per essere utilizz...

<ol>
	<li>Robinson, G. E., Fernald, R. D., Clayton, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genes+and+social+behavior.">Genes and social behavior.</a> Science. 322, 896-900 (2008).</li><li>Koob, G. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurobiological+substrates+for+the+dark+side+of+compulsivity+in+addiction.">Neurobiological substrates for the dark side of compulsivity in addiction.</a> Neuropharmacol. 56, 18-31 (2009).</li><li>Kaun, K. R., Devineni, A. V., Heberlein, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+melanogaster+as+a+model+to+study+drug+addiction.">Drosophila melanogaster as a model to study drug addiction.</a> Hum Genet. 131, 959-975 (2012).</li><li>Devineni, A. V., Heberlein, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Addiction-like+behavior+in+Drosophila.">Addiction-like behavior in Drosophila.</a> Commun Integr Biol. 3, 357-359 (2010).</li><li>Shohat-Ophir, G., Kaun, K. R., Azanchi, R., Mohammed, H., Heberlein, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+deprivation+increases+ethanol+intake+in+Drosophila.">Sexual deprivation increases ethanol intake in Drosophila.</a> Science. 335, 1351-1355 (2012).</li><li>Devineni, A. V., Heberlein, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preferential+ethanol+consumption+in+Drosophila+models+features+of+addiction.">Preferential ethanol consumption in Drosophila models features of addiction.</a> Curr Biol : CB. 19, 2126-2132 (2009).</li><li>McBride, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mushroom+body+ablation+impairs+short-term+memory+and+long-term+memory+of+courtship+conditioning+in+Drosophila+melanogaster.">Mushroom body ablation impairs short-term memory and long-term memory of courtship conditioning in Drosophila melanogaster.</a> Neuron. 24, 967-977 (1999).</li><li>Ejima, A., Smith, B. P., Lucas, C., Levine, J. D., Griffith, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sequential+learning+of+pheromonal+cues+modulates+memory+consolidation+in+trainer-specific+associative+courtship+conditioning.">Sequential learning of pheromonal cues modulates memory consolidation in trainer-specific associative courtship conditioning.</a> Curr Biol. 15, 194-206 (2005).</li><li>Ejima, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generalization+of+courtship+learning+in+Drosophila+is+mediated+by+cis-vaccenyl+acetate.">Generalization of courtship learning in Drosophila is mediated by cis-vaccenyl acetate.</a> Curr Biol. 17, 599-605 (2007).</li><li>Keleman, K., Kruttner, S., Alenius, M., Dickson, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Function+of+the+Drosophila+CPEB+protein+Orb2+in+long-term+courtship+memory.">Function of the Drosophila CPEB protein Orb2 in long-term courtship memory.</a> Nat Neurosci. 10, 1587-1593 (2007).</li><li>Kaun, K. R., Azanchi, R., Maung, Z., Hirsh, J., Heberlein, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Drosophila+model+for+alcohol+reward.">A Drosophila model for alcohol reward.</a> Nat Neurosci. 14, 612-619 (2011).</li><li>Xu, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+propensity+for+consuming+ethanol+in+Drosophila+requires+rutabaga+adenylyl+cyclase+expression+within+mushroom+body+neurons.">The propensity for consuming ethanol in Drosophila requires rutabaga adenylyl cyclase expression within mushroom body neurons.</a> Genes Brain Behav. 11, 727-739 (2012).</li><li>Devineni, A. V., Heberlein, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+evolution+of+Drosophila+melanogaster+as+a+model+for+alcohol+research.">The evolution of Drosophila melanogaster as a model for alcohol research.</a> Annu Rev Neurosci. 36, 121-138 (2013).</li><li>Ro, J., Harvanek, Z. M., Pletcher, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FLIC:+high-throughput,+continuous+analysis+of+feeding+behaviors+in+Drosophila.">FLIC: high-throughput, continuous analysis of feeding behaviors in Drosophila.</a> PloS one. 9, 101107 (2014).</li><li>Itskov, P. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+monitoring+and+quantitative+analysis+of+feeding+behaviour+in+Drosophila.">Automated monitoring and quantitative analysis of feeding behaviour in Drosophila.</a> Nat Commun. 5, 4560 (2014).</li><li>Yapici, N., Cohn, R., Schusterreiter, C., Ruta, V., Vosshall, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Taste+Circuit+that+Regulates+Ingestion+by+Integrating+Food+and+Hunger+Signals.">A Taste Circuit that Regulates Ingestion by Integrating Food and Hunger Signals.</a> Cell. 165, 715-729 (2016).</li><li>Peru, Y. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-lasting,+experience-dependent+alcohol+preference+in+Drosophila.">Long-lasting, experience-dependent alcohol preference in Drosophila.</a> Addict Biol. 19, 392-401 (2014).</li><li>Dus, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nutrient+Sensor+in+the+Brain+Directs+the+Action+of+the+Brain-Gut+Axis+in+Drosophila.">Nutrient Sensor in the Brain Directs the Action of the Brain-Gut Axis in Drosophila.</a> Neuron. 87, 139-151 (2015).</li><li>Anderson, D. J., Adolphs, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+framework+for+studying+emotions+across+species.">A framework for studying emotions across species.</a> Cell. 157, 187-200 (2014).</li><li>Gibson, W. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+responses+to+a+repetitive+visual+threat+stimulus+express+a+persistent+state+of+defensive+arousal+in+Drosophila.">Behavioral responses to a repetitive visual threat stimulus express a persistent state of defensive arousal in Drosophila.</a> Curr Biol. 25, 1401-1415 (2015).</li></ol>

Here, we illustrate the details of an integrated approach to measure alterations in reward-seeking behavior, based on previous work described by Devineni et al.6 and Shohat-Ophir et al5. The first section of the protocol uses different types of sexual interactions as the experience input, and the second section uses a two-choice feeding assay to assess the effect of experience on the preference to consume ethanol.
As shown by Devineni et al.<sup...

Modifica del comportamento in risposta alle esperienze permette animali di regolare il loro comportamento ai cambiamenti nel loro ambiente 1. Durante questo processo, gli animali integrano il loro stato fisiologico interno con mutevoli condizioni dell&#39;ambiente esterno e successivamente scegliere un&#39;azione piuttosto che un altro per aumentare le loro possibilità di sopravvivenza e la riproduzione. Sistemi di ricompensa si sono evoluti per motivare i comportamenti che sono necessari per la sopravvivenza di individui e di specie rafforzando i comportamenti che aumentano la sopravvivenza immediata, come mangiare o bere, o quelli che garantiscono la sopravvivenza a lungo termine, come ad esempio il comportamento sessuale o la cura per la prole 2. Composti artificiali, come l&#39;abuso di droghe interessano anche sistemi di ricompensa per vie neurali cooptazione che mediano ricompense naturali 2. Nel corso degli ultimi due decenni, la mosca della frutta Drosophila melanogaster è stato stabilito come un modello promettente per lo studio della Moleclare e dei meccanismi neuronali che stanno plasmando gli effetti di etanolo sul comportamento 3,4. In precedenza, abbiamo identificato un sottogruppo di neuroni peptidergiche nei moscerini (recettore NPF / NPF (R) neuroni) che premia naturali coppia, come esperienza sessuale, alla motivazione di ottenere ricompense droga 5. espressione NPF è sensibile ad entrambe le esperienze sessuali e alle ricompense di droga, come l&#39;etanolo intossicazione. I cambiamenti nei livelli di espressione NPF vengono convertiti in etanolo alterazioni autosomministrazione 5, dove l&#39;alta NPF riduce e bassa NPF aumenta la preferenza a consumare etanolo. L&#39;attivazione dei neuroni NPF è gratificante per le mosche, in quanto mostrano una forte preferenza per un odore in coppia con l&#39;attivazione, che si riflette anche dalla riduzione del consumo di etanolo. Ancora più importante, l&#39;attivazione dei neuroni NPF interferisce con la capacità di mosche formare un&#39;associazione positiva tra etanolo intossicazione e un cue odore. Il nesso causale tra la NPF / Rsistema, la memoria di ricompensa, e il consumo di etanolo suggerisce che si può usare l&#39;etanolo autosomministrazione come misura per le modifiche negli stati ricompensa 5. In questa pubblicazione abbiamo dimostrato un approccio integrato per attingere il sistema di ricompensa naturale mosca e modifiche del test negli stati di ricompensa. L&#39;approccio è costituito da due parti separate, un protocollo di formazione per la manipolazione di esperienze naturali ricompensa legati, seguita da una a due scelta saggio alimentatore capillare (CAFE) per valutare l&#39;etanolo autosomministrazione come una stima per i cambiamenti negli stati di ricompensa. Il saggio CAFE è analogo ai saggi scelta due bottiglie utilizzate in studi su roditori drug autosomministrazione ed è stato dimostrato per riflettere alcune proprietà di comportamento dipendenza simile mosche 6.

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a simple way to measure alterations in reward-seeking behavior using drosophila melanogaster

Descriviamo un protocollo per misurare l&#39;etanolo autosomministrazione nei moscerini della frutta (Drosophila melanogaster) come proxy per i cambiamenti negli stati di ricompensa. Dimostriamo un modo semplice per sfruttare il sistema di ricompensa volo, modificare le esperienze legate alla ricompensa naturale, e utilizzare consumo di etanolo volontaria come misura per le modifiche negli stati di ricompensa. L&#39;approccio è uno strumento rilevante per studiare i neuroni e geni che svolgono un ruolo nella esperienza cambia-mediata di stato interno. Il protocollo si compone di due parti distinte: esporre le mosche di esperienze gratificanti e nonrewarding, e analizzando il consumo di etanolo volontariato come una misura della motivazione per ottenere un premio di droga. Le due parti possono essere usati indipendentemente per indurre la modulazione di esperienza come primo passo per ulteriori analisi a valle o come dosaggio alimentazione due scelta indipendenti, rispettivamente. Il protocollo non richiede una configurazione complessa e quindi può essere applicato in qualsiasi laboratory con strumenti di base della cultura mosca.

FPreviously, Devineni et al. hanno dimostrato che quando i moscerini della frutta sono dati la scelta di consumare cibo contenente etanolo, mostrano una forte preferenza per il cibo contenente etanolo sopra nonethanol contenente cibo 6. Indicato qui ci sono alcuni risultati rappresentativi si ottengono quando saggiare la preferenza etanolo innata di mosche maschio ingenua che non ha subito il protocollo di allenamento. <p class="jove_content" fo:keep-together.with...

Watch this Scientific Journal Video about A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster at JoVE.com

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

Descriviamo un protocollo per indurre gratificante e nonrewarding esperienze nei moscerini della frutta (Drosophila melanogaster) utilizzando il consumo di etanolo volontaria come misura per le modifiche negli stati di ricompensa.

Un modo semplice per misurare alterazioni in ricompensa il comportamento di ricerca Utilizzando<em&gt; Drosophila melanogaster</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

The overall goal of this procedure is to measure ethanol self-administration in fruit flies as a proxy for changes in rewards states. This method can help answer key questions in neuroscience, such as, how experience is encoded in the brain, and leads to modification of behavior. The main advantage of this technique is that it is relatively simple, and does not require a complicated setup.The protocol is composed of two discrete parts. The first is exposing the flies to rewarding and non-rewarding experiences, and the second is assaying voluntary ethanol consumption as a measure of the motivation to obtain a drug reward. The two parts of the protocol can be used independently.To induce the modulation of experience as an initial step for further downstream assays, or, as an independent, two-choice feeding assay. Demonstrating the procedure will be Shir Zer, a master's student, and Julia Ryvkin, a graduate student, both from my lab. To begin the construction of the capillary feeder, first take a standard fly vial with small holes along the side of the vial.Use a razor blade to cut the plug of the vial in half, crosswise. Mark four points onto the surface of the plug, creating a square shape to position the capillary holders. With an 18-gauge needle, make four holes through the plug at the position of the marks.And make the holes wider using a micropipette. Using a razor blade, cut four micropipettes so that they firmly hold five microliter glass capillaries. Insert these into the holes in the prepared plug.Mark the position on the plug of the two capillaries that will contain ethanol. Next, arrange glass vials in microcentrifuge racks. Melt fly food, and then pour two milliliters into each of the vials.Allow the food to harden, and then cover the vials with plastic wrap. Begin by taking male flies that have been housed separately in smaller vials in seclusion. Assign these to two groups randomly.Those who will undergo rewarding, versus non-rewarding mating experiences. Beginning with the non-rewarding group, add a previously mated female to each vial. Moving on to the reward group, add a virgin female to each vial.Watch the mating encounters to ensure that males interacting with virgin females mate successfully within the first hour. And that males interacting with previously mated females do not mate. It is essential to observe carefully, to make sure that the mated cohort do indeed undergo mating, and that the rejected cohort do not manage to mate.After one hour, remove from the experiment any males from the rejected cohort that managed to mate, and males from the mated cohort that did not end up mating by the end of the first training session. Conclude the conditioning session by aspirating out the previously mated females from the non-rewarding condition, keeping these individuals for later trials. Finally, remove and discard the females from the mated male group.Let the flies rest for one hour, and then repeat the hour long conditioning assay two further times with an hour rest in between. These training sessions should be repeated again each day for three more days. After the final conditioning session on day four, aspirate the single males and place them in groups of six to eight into the capillary feeder vials, keeping flies in the same reward group together.Use a soft plug to keep the males in place during transfer. Once all flies are inserted, replace the soft plug with the capillary feeder plug. Wet the plugs by gently adding five milliliters of water to the top of each plug, avoiding spillage into the vial.Prepare the ethanol and water food solutions, and then place five microliter drops of these onto a strip of laboratory film. Allow the solution to fill the capillary until it reaches the five microliter mark. And then tap them gently on the strip to position the solution exactly at the premarked line.Seal the ends of the capillaries by dipping them into small droplets of mineral oil. Finally, insert the two ethanol containing and non-ethanol containing capillaries into the adapters in the plug. The exposed sides should protrude one millimeter or less from the plug surface.Keeping the opening for the capillaries close to the wet plug reduces the evaporation and allows easy access to the food, as flies normally sit on the plugs and climb down on the exposed capillary to feed. Additionally, prepare one mock vial without flies, which will be used as a control to assess natural evaporation. Finally, record the relative position of the liquid in each of the capillaries in vials with respect to the black line in millimeters.Set the vials into the experimental incubator for 24 hours. Measure and record the levels of the liquid in each removed capillary with respect to the black line. Once the experiment is in progress, replace the capillary at a set time, each day.Re-dampen the plugs with two milliliters of water, and then set the vials back into the incubator. Wild type Canton S strain male fruit flies collected upon eclosion and aged until four days old, were assayed over the course of four days for their innate preference to consume alcohol. Analysis of volumes consumed from ethanol and non-ethanol containing solutions, show that the flies exhibited a significant preference for the 15%ethanol food source.The CAFE experiment was repeated with mated versus rejected males. Both groups had been previously housed singly, and exposed to females that either accepted or rejected mating. Upon introduction in groups to the CAFE vials, the rejected group consumed more from the ethanol capillaries.This suggested that perceived lack of reward led to an increase in reward-seeking behavior. Conversely, successful mating increases internal reward levels, which in turn lowers ethanol consumption. While attempting this procedure, it's important to maintain a well-humidified environment for the entire experiment, with extra emphasis on the consumption part, watering the plugs every day, and keeping the capillary ends in close proximity to the wet plugs.

Research

JoVE Journal

Neuroscience

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

Magnetic Resonance Spectroscopy of live Drosophila melanogaster using Magic Angle Spinning

Magnetic Resonance Spectroscopy of live Drosophila melanogaster using Magic Angle Spinning

Spettroscopia di Risonanza Magnetica dal vivo Drosophila melanogaster Utilizzando Magic Angle Spinning

High-Resolution Magic Angle Spinning (HRMAS) proton magnetic resonance spectroscopy (1H-MRS) is a novel non-destructive technique that improves spectral ...

Ricerca

Neuroscienze

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Registrazioni Sensillum singolo in Insetti Drosophila melanogaster E 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 ...

A Simple Way to Measure Ethanol Sensitivity in Flies

Un modo semplice per misurare sensibilità etanolo in Flies

Low doses of ethanol cause flies to become hyperactive, while high doses are sedating. The sensitivity to ethanol-induced sedation of a given fly strain ...

Simultaneous Recording of Calcium Signals from Identified Neurons and Feeding Behavior of Drosophila melanogaster

Simultaneous Recording of Calcium Signals from Identified Neurons and Feeding Behavior of Drosophila melanogaster

La registrazione simultanea dei segnali di calcio da parte dei neuroni identificati e comportamento alimentare di Drosophila melanogaster</em

To study neuronal networks in terms of their function in behavior, we must analyze how neurons operate when each behavioral pattern is generated.  Thus, ...

Methods to Assay Drosophila Behavior

Methods to Assay Drosophila Behavior

Metodi per test Drosophila Comportamento

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular ...

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Stimolazione Optogenetic del comportamento in Fuga<em&gt; Drosophila melanogaster</em

A  growing number of genetically encoded tools are becoming available that allow non-invasive  manipulation of the neural activity of specific neurons in ...

Conditions Affecting Social Space in Drosophila melanogaster

Conditions Affecting Social Space in Drosophila melanogaster

Condizioni che influiscono spazio sociale in<em&gt; Drosophila melanogaster</em

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a ...

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

Ad alta risoluzione quantificazione del comportamento Odor-guidata<em&gt; Drosophila melanogaster</em&gt; Utilizzo della<em&gt; Flywalk</em&gt; Paradigm

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To ...

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

Utilizzando canali lineari Agarose per studiare<em&gt; Drosophila</em&gt; Comportamento larvale Crawling

Drosophila larval crawling is emerging as a powerful model to study neural control of sensorimotor behavior. However, larval crawling behavior on flat ...

Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons

Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons

Il comportamento larvale della drosofila in risposta alla stimolazione optogenetica dei neuroni olfattivi di rilevamento

The ability of insects to navigate toward odor sources is based on the activities of their first-order olfactory receptor neurons (ORNs). While a ...