Biz el yazması geliştirmek için kendi iş için 4 anonim yorumcular teşekkür ederim. Biz MBD ve YG için mali destek için Centre National de la Recherche SCIENTIFIQUE teşekkür ve MMS Université de Bourgogne ve Araştırma Bakanlığı Fransız. YG Araştırma laboratuvarında Avrupa Araştırma Konseyi (ERC Grant, GliSFCo-311403 Başlangıç), Agence Nationale de la Recherche (ANR-JCJC, GGCB-2010), Conseil Régional de Bourgogne (Faber) ve CNRS tarafından finanse edilmektedir.

1.. Başlamadan Önce <ol><li> Istikrarlı ve sağlam davranışsal fenotipleri taşıyan bir isogenized referans stok kullanın. Tüm potansiyel kontrolleri heterojen plan alellerini taşıyabilir, çünkü bu stok seçimi için genel bir kural yoktur. </li><li> Sonraki adımlar için gerekli olan her stok geri çaprazlama için bu kontrol suşu kullanın. Bu adım, tipik olarak 2-3 geriye dönük melezleştirme izogenik referans erkeklerde 5 (mümkün çaprazlama homolog kromozomlar arasındaki izin vermek için), tek bir bakire dişi, en az 5 ardışık çarpılar ile temsil edilir. Bu adım, farklı sinek stoklarının arasındaki genetik arka planı homojenize edilmesi önemlidir. </li><li> 25 ° C&#39;de 12 saat ışık / karanlık döngüsünde antibiyotik (% 0.4 metil para-hidroksi-benzoat) ile tamamlanır standart mısır unu, (% 9), maya (% 10) ve agar ortamında (% 1.5) üzerinde Drosophila stokları korumak ° C. </li><li> Sıcaklık-kontrollü bir odaya uzak kırmızı ışık altında (25 ° C) içerisinde kimyasal-duyusal deneyler elde(Görsel ipuçlarının katkısını ortadan kaldırmak için, ve kimyasal duyusal sinyallere odaklanmak). Düzenli olarak her deney arasındaki bölgeyi havalandırmak için odanın havasını yenilemek. </li></ol> Bir Y-labirenti Assay kullanılarak, 2. Koku Response <ol><li> Test edilmeden önce ıslak kağıt havlu ihtiva eden cam tüpler içinde 25 ° C&#39;de 16-18 saat boyunca sinekler açlıktan. </li><li> İki cam şişeler için ve daha küçük plastik bir şişe (şişe yükleme) bir Y-şekilli konektör tarihi. Üç şişelerine konektörü bağlantı ve sıkıca kapatılmış bir Y-labirent elde etmek için köpük stoper geçmesine 1 ml pipet ipuçlarını kullanın. İki &quot;tuzak&quot; şişeleri oluşturmak için, ve bir pipet büyük bir ucu &quot;yükleme&quot; oluşturmak için (onun kararını sonra sinek herhangi bir geri dönüş önlemek için ~ 2 mm çap) iki pipet ipuçları dar uçlarını kesin boru (Şekil 1A). </li><li> Sadece &quot;tuzak&quot; şişeleri (Şekil 1B) bağlamadan önce, bir ~ 6 mm çaplı filtre kağıdı yerleştirinHer flakon. Bir filtre kağıdı üzerinde odorant çözeltisi 40 ul, ikinci filtre kağıdı üzerinde karşılık gelen çözücünün 40 ul ekle. </li><li> &quot;Yükleme&quot; şişenin içine on 4-9 günlük eski sinekler tanıtmak. Bu davranış 10 üzerinde güçlü bir etkiye sahip olduğundan, bu transfer sırasında CO 2 anestezi kullanmayın. Aksine buz üzerinde kısa soğutma kullanın. Anestezi sinekler doğru manipülasyonu mümkün olduğunca konularda baskısını sınırlandırmak için önemlidir. <ol><li> Görsel uyaranlara mümkün olduğunca kaçınmak için (mümkünse ısı kaynağı sınırlamak için LED ampuller kullanarak) uzak-kırmızı ışık altında, 25 ° C&#39;de Y-labirent bir dizi test gerçekleştirin. (; Şekil 1C solda veya sağda Odorant içeren tüp ve yükleme tüp ön veya arka) Y-labirentine yönelimleri alternatif için dikkatli olun. </li><li> Sinekler için birkaç saat odorant ya da çözücüyü içeren tuzak şişede girmek izin verin. Participati geliştirmek için 24 saat sonra, sinekler Toplamıfazla% 80 kadar ve maksimum koku indeks değeri (Simonnet, kişisel iletişim) sağlamak. </li><li> - / Yüklü sineklerin sayısı (solvent tüp numara koku tüp içinde sayı): aşağıdaki formül kullanılarak elde edilen koku indeksi hesaplayın. </li></ol></li><li> Yıkama Y-labirent set-up şöyle: gecede RBS 35 MD sökülmüş set-up ıslatın. Iyice musluk suyu ile yıkayın. Sonunda deiyonize su ile durulayın ve kurumasını. </li></ol> Verilerin 3. İstatistiksel Analiz <ol><li> Veriler ve çeşitli değişkenlere bağlı olarak, bir t-testi, tek-yönlü ANOVA ya da bir iki-yönlü ANOVA gerçekleştirin. </li></ol>

<ol>
	<li>Mombaerts, P. <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+ligands+for+odorant,+vomeronasal+and+taste+receptors.">Genes and ligands for odorant, vomeronasal and taste receptors.</a> Nat. Rev. Neurosci. 5, 263-278 (2004).</li><li>Silbering, A. F., Benton, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ionotropic+and+metabotropic+mechanisms+in+chemoreception:+'chance+or+design.">Ionotropic and metabotropic mechanisms in chemoreception: 'chance or design.</a> EMBO Rep. 11, 173-179 (2010).</li><li>Ziegler, A. B., Berthelot-Grosjean, M., Grosjean, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+smell+of+love+in+Drosophila.">The smell of love in Drosophila.</a> Front. Physiol. 4, (2013).</li><li>Silbering, A. F., 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=Complementary+function+and+integrated+wiring+of+the+evolutionarily+distinct+Drosophila+olfactory+subsystems.">Complementary function and integrated wiring of the evolutionarily distinct Drosophila olfactory subsystems.</a> J. Neurosci. 31, 13357-13375 (2011).</li><li>Grosjean, Y., 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=An+olfactory+receptor+for+food-derived+odours+promotes+male+courtship+in+Drosophila.">An olfactory receptor for food-derived odours promotes male courtship in Drosophila.</a> Nature. 478, 236-240 (2011).</li><li>Woodard, C., Huang, T., Sun, H., Helfand, S. L., Carlson, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+analysis+of+olfactory+behavior+in+Drosophila:+a+new+screen+yields+the+ota+mutants.">Genetic analysis of olfactory behavior in Drosophila: a new screen yields the ota mutants.</a> Genetics. 123, 315-326 (1989).</li><li>Semmelhack, J. L., Wang, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Select+Drosophila+glomeruli+mediate+innate+olfactory+attraction+and+aversion.">Select Drosophila glomeruli mediate innate olfactory attraction and aversion.</a> Nature. 459, 218-223 (2009).</li><li>Budick, S. A., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Free-flight+responses+of+Drosophila+melanogaster+to+attractive+odors.">Free-flight responses of Drosophila melanogaster to attractive odors.</a> The Journal of Experimental Biology. 209, 3001-3017 (2006).</li><li>Martin, F., Charro, M. J., Alcorta, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mutations+affecting+the+cAMP+transduction+pathway+modify+olfaction+in+Drosophila.">Mutations affecting the cAMP transduction pathway modify olfaction in Drosophila.</a> J. Comp. Physiol. A. 187, 359-370 (2001).</li><li>Barron, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anaesthetising+Drosophila+for+behavioural+studies.">Anaesthetising Drosophila for behavioural studies.</a> Journal of Insect Physiology. 46, 439-442 (2000).</li><li>Ai, 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=Acid+sensing+by+the+Drosophila+olfactory+system.">Acid sensing by the Drosophila olfactory system.</a> Nature. 468, 691-695 (2010).</li><li>Becher, P. G., Bengtsson, M., Hansson, B. S., Witzgall, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flying+the+fly:+long-range+flight+behavior+of+Drosophila+melanogaster+to+attractive+odors.">Flying the fly: long-range flight behavior of Drosophila melanogaster to attractive odors.</a> J. Chem. Ecol. 36, 599-607 (2010).</li></ol>

Yazarlar ifşa hiçbir şey yok.

Bizim Y-labirenti protokol, önceden tanımlanmış bir protokole 9 dayanmaktadır. Ancak, iki önemli farklılıklar tanıtmak. İlk olarak, bu çözücü veya çözücü artı odorant ihtiva eden şişeye girmek için karar bir kez geri gelen sinekleri önlemek için dar bir pipet uçları kullanın. Bu dar ipuçları da Y-labirentte odorant difüzyon sınırlamak için yararlıdır. İkinci olarak, Y-labirent girmek için sinekleri zorlamak için daha küçük bir yükleme şişeyi kullanın. Bu sinekler b...

Tat veya koku duyusunun aracılığıyla kemo-alış hayvan hayatta kalmak için önemli bir duyusal yöntemidir. Bu hayati bir tehlike veya gıda kaynaklarının tespiti için gerekli ipuçlarını, yanı sıra sosyal etkileşimleri için verir. Aynı zamanda onların üreme için gerekli bir seks partneri bulmak için hayvanlar olur. 20 yılı aşkın için, Nobel ödülü &quot;odorant reseptörleri ve koku alma sisteminin organizasyonunun buluşları için&quot; 2004 yılında Richard Axel ve Linda Buck tarafından çalışma kazanan dahil olmak üzere yoğun araştırma, moleküler ve hücresel üsleri ortaya çıkarmak için yapılmıştır olfaction 1,2. Koku algısını incelemek için bilim adamları için favori hayvan modellerinden biri olan D. melanogaster. Bu böcek memeliler ile benzer hücresel ve moleküler koku kodlama stratejisi paylaşıyor. Bilimsel topluluk bu meyve sineği koku rolünü incelemek için çeşitli davranış paradigmaları kullanır. Bu testler, çok modlu courtshi gibi tahliller bulunmaktadırolfaction dahil olmak üzere çeşitli duyu modaliteleri, erkek iltifata 3 ortaya çıkarmak için önemli olan p testleri. Diğer deneyler, aynı zamanda daha özel olarak koku rolünü mücadele için geliştirilmiştir; bu T-labirentine, Y-labirentine, tuzak deneyleri, dört alan arenalarda ve rüzgar tünelleri 4,5,6,7,8 içerir. Bu yazıda D. kullanarak sağlam koku yanıtları sağlayan basit bir modifiye Y-labirent testi, mevcut melanogaster. Bizim set-up daha önce açıklanan yönteme 9 aksine son ipuçlarını kullanır. Böylece, Y-labirenti iki avantajı vardır. Sinek kendi seçimini yapmış kez Birincisi, bu sistemde herhangi bir geri dönüş önler. İkinci olarak, Y-labirent tüm alanlarında koku değişimini sınırlar. Drosophila genellikle odorant doymasını önlemek için kullanılan hava akımına karşı çok hassas olduğu için, bu son avantaj önemlidir. Bir hava akışı ile deneysel set-up ayarlamak için zaman ve maliyet alıcı olacaktır. Bu nedenle, Y-labirenti deneyi temsil efşır ve Drosophila koku performansını test etmek için hızlı yolu.

<table border="0" cellpadding="0" cellspacing="0" width="835">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr>
			<td>Drosophila Polystyrene tube</td>
			<td>VWR europe</td>
			<td>734-2255</td>
			<td>30 x 25 mm&#160; 
			Y-maze</td>
		</tr>
		<tr>
			<td>Drosophila Borosilicate tube</td>
			<td>Dijon verre</td>
			<td></td>
			<td>95 X 25 mm 
			Y-maze</td>
		</tr>
		<tr>
			<td>Foam stopper</td>
			<td>Dutscher</td>
			<td>999038</td>
			<td> 
			Y-maze</td>
		</tr>
		<tr>
			<td>Y-shaped connector</td>
			<td>Europrix</td>
			<td>11020605</td>
			<td> 
			Y-maze</td>
		</tr>
		<tr>
			<td>100-1000&#181;l pipet tips</td>
			<td>Corning</td>
			<td>4868</td>
			<td>Join the following pipet tips to the Y-shaped connector. Cut 2 pipet tips at 65 mm from the wide end, and connect the narrow end (with a &#8764;2 mm opening) to 2 test vials. These openings will limit the U-turns once the flies enter the tubes containing the odors. Cut 1 pipet tip at 35 mm from the wide end, and connect it to the loading vial. 
			Y-maze</td>
		</tr>
		<tr>
			<td>Far-Red LED Bulb</td>
			<td>Rubin-Lacaque</td>
			<td>0RB180238</td>
			<td>625-630 nm</td>
		</tr>
		<tr>
			<td>Acetic Acid</td>
			<td>Sigma-Aldrich</td>
			<td>45725</td>
			<td></td>
		</tr>
		<tr>
			<td>Phenylacetic Acid</td>
			<td>Sigma-Aldrich</td>
			<td>P16621</td>
			<td></td>
		</tr>
		<tr>
			<td>Yeast</td>
			<td>Sensient Flavors Strasbourg</td>
			<td>1018880464</td>
			<td></td>
		</tr>
		<tr>
			<td>Cornmeal</td>
			<td>eurogerm</td>
			<td>Farine de ma&#239;s</td>
			<td></td>
		</tr>
		<tr>
			<td>Agar</td>
			<td>Kalys</td>
			<td>HP-697-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Methyl hydroxy 4 benzoate</td>
			<td>VWR international</td>
			<td>25605293</td>
			<td></td>
		</tr>
	</tbody>
</table>

testing drosophila olfaction with a y-maze assay

Hayvanlar onların hayatta kalmasını sağlamak için çevreden sinyalleri tespit esastır. Bu amaçla, onlar doğrudan temas yoluyla veya daha uzun mesafeden etkili bir uçucu moleküle yanıtı temsil olfaction, yoluyla, bu vizyon, mechanoreception, işitme ve tat ile chemoperception gibi çevresel ipuçlarını kullanın. Uçucu kimyasal moleküller tehlike, bir besin kaynağı, ya da bireyler arasında iletişim kurmak için saptanmasında çok hayvanlar için çok önemli sinyallerdir. Drosophila melanogaster olfaction hücresel ve moleküler temeli keşfetmek bilim adamları için en yaygın biyolojik modelleri biridir. Bu küçük böcek koku yeteneklerini vurgulamak amacıyla, klasik fareler ile birlikte kullanılan Y-labirenti testine göre bir tadil edilmiş bir seçim protokol açıklar. Y-labirentine ile elde edilen veriler daha iyi hayvanlar sürekli değişen çevre ile nasıl başa anlamak için değerli bilgiler verir. Biz çalışmak için bir adım-adım protokolü tanıtmakBu Y-labirent testi kullanılarak sinek keşif cevabına koku etkisi.

Şekil 1B, bu Y-labirenti tahlili kullanılarak iki temsili tepkilerini gösterir. Önemli ölçüde% 10 fenilasetik asit kaçının oysa Canton-S erkek güçlü, damıtılmış su içerisinde seyreltilmiş,% 10 asetik asit kaçının. Bu deneyler yükleme şişeler içinde bir araya geldiklerinde her tekrar için 10 erkek dayanmaktadır. Bu protokol, bazen büyük ortalama standart hata yol açabilir. Gerekirse, bunun yerine, sadece 10. Matematiksel olarak, bir bireyin seçimi daha büyük bir numune s...

Watch this Scientific Journal Video about Testing Drosophila Olfaction with a Y-maze Assay at JoVE.com

Testing Drosophila Olfaction with a Y-maze Assay

Biz Y-labirent testi kullanılarak Drosophila davranışı üzerindeki koku etkisini ortaya çıkarmak için bir seçim testi sunuyoruz.

Test<em&gt; Drosophila</emBir Y-labirenti testi ile&gt; koku alma

Detecting signals from the environment is essential for animals to ensure their survival. To this aim, they use environmental cues such as vision, ...

testing-drosophila-olfaction-with-a-y-maze-assay

Research

JoVE Journal

Neuroscience

Testing Drosophila Olfaction with a Y-maze Assay

14.8K Görüntüleme.  UMR-6265 CNRS.  Biz Y-labirent testi kullanılarak Drosophila davranışı üzerindeki koku etkisini ortaya çıkarmak için bir seçim testi sunuyoruz. 

Video: Testing Drosophila Olfaction with a Y-maze Assay

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

Screening compounds for in vivo activity can be used as a first step to identify candidates that may be developed into pharmacological agents1,2. We developed a novel nanoinjection/electrophysiology assay that allows the detection of bioactive modulatory effects of compounds on the function of a neuronal circuit that mediates the escape response in Drosophila melanogaster3,4. Our in vivo assay, which uses the Drosophila Giant Fiber System (GFS, Figure 1) allows screening of different types of compounds, such as small molecules or peptides, and requires only minimal quantities to elicit an effect. In addition, the Drosophila GFS offers a large variety of potential molecular targets on neurons or muscles. The Giant Fibers (GFs) synapse electrically (Gap Junctions) as well as chemically (cholinergic) onto a Peripheral Synapsing Interneuron (PSI) and the Tergo Trochanteral Muscle neuron (TTMn)5. The PSI to DLMn (Dorsal Longitudinal Muscle neuron) connection is dependent on D&alpha;7 nicotinic acetylcholine receptors (nAChRs)6. Finally, the neuromuscular junctions (NMJ) of the TTMn and the DLMn with the jump (TTM) and flight muscles (DLM) are glutamatergic7-12. Here, we demonstrate how to inject nanoliter quantities of a compound, while obtaining electrophysiological intracellular recordings from the Giant Fiber System13 and how to monitor the effects of the compound on the function of this circuit. We show specificity of the assay with methyllycaconitine citrate (MLA), a nAChR antagonist, which disrupts the PSI to DLMn connection but not the GF to TTMn connection or the function of the NMJ at the jump or flight muscles.
Before beginning this video it is critical that you carefully watch and become familiar with the JoVE video titled "Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster " from Augustin et al7, as the video presented here is intended as an expansion to this existing technique. Here we use the electrophysiological recordings method and focus in detail only on the addition of the paired nanoinjections and monitoring technique.

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

A rapid in vivo assay to test for neuromodulatory compounds using the Giant Fiber System (GFS) of Drosophila melanogaster is described. Nanoinjections in the head of the animal along with electrophysiological recordings of the GFS can reveal bioactivity of compounds on neurons or muscles.

Kullanarak Bileşiklerin Biyoaktivite için eşli Nanoinjection ve Ekran için Elektrofizyoloji Testi Drosophila melanogaster Dev Elyaf Sistemi

Screening compounds for in vivo activity can be used as a first step to identify candidates that may be developed into pharmacological agents1,2. We ...

Nörobilim

Methods to Assay Drosophila Behavior

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular disease and various neurological diseases1. We have optimized simple and robust behavioral assays for determining larval locomotion, adult climbing ability (RING assay), and courtship behaviors of Drosophila. These behavioral assays are widely applicable for studying the role of genetic and environmental factors on fly behavior. Larval crawling ability can be reliably used for determining early stage changes in the crawling abilities of Drosophila larvae and also for examining effect of drugs or human disease genes (in transgenic flies) on their locomotion. The larval crawling assay becomes more applicable if expression or abolition of a gene causes lethality in pupal or adult stages, as these flies do not survive to adulthood where they otherwise could be assessed. This basic assay can also be used in conjunction with bright light or stress to examine additional behavioral responses in Drosophila larvae. Courtship behavior has been widely used to investigate genetic basis of sexual behavior, and can also be used to examine activity and coordination, as well as learning and memory. Drosophila courtship behavior involves the exchange of various sensory stimuli including visual, auditory, and chemosensory signals between males and females that lead to a complex series of well characterized motor behaviors culminating in successful copulation. Traditional adult climbing assays (negative geotaxis) are tedious, labor intensive, and time consuming, with significant variation between different trials2-4. The rapid iterative negative geotaxis (RING) assay5 has many advantages over more widely employed protocols, providing a reproducible, sensitive, and high throughput approach to quantify adult locomotor and negative geotaxis behaviors. In the RING assay, several genotypes or drug treatments can be tested simultaneously using large number of animals, with the high-throughput approach making it more amenable for screening experiments.

Methods to Assay Drosophila Behavior

Drosophila melanogaster is a genetically and behaviorally tractable model system that has been used to understand the molecular and cellular basis of many important biological processes for over a century 1. Drosophila has been well exploited to gain insights into the genetic basis of fly behavior.

Test için Yöntemler Drosophila Davranış

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

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

Glucose metabolism is a useful marker for local neural activity, forming the basis of methods such as 2-deoxyglucose and functional magnetic resonance imaging. However, use of such methods in animal models requires anesthesia and hence both alters the brain state and prevents behavioral measures. An alternative method is the use of in vivo microdialysis to take continuous measurement of brain extracellular fluid concentrations of glucose, lactate, and related metabolites in awake, unrestrained animals. This technique is especially useful when combined with tasks designed to rely on specific brain regions and/or acute pharmacological manipulation; for example, hippocampal measurements during a spatial working memory task (spontaneous alternation) show a dip in extracellular glucose and rise in lactate that are suggestive of enhanced glycolysis1-3,4-5, and intrahippocampal insulin administration both improves memory and increases hippocampal glycolysis6. Substances such as insulin can be delivered to the hippocampus via the same microdialysis probe used to measure metabolites. The use of spontaneous alternation as a measure of hippocampal function is designed to avoid any confound from stressful motivators (e.g. footshock), restraint, or rewards (e.g. food), all of which can alter both task performance and metabolism; this task also provides a measure of motor activity that permits control for nonspecific effects of treatment. Combined, these methods permit direct measurement of the neurochemical and metabolic variables regulating behavior.

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

Modulation of hippocampally-dependent spatial working memory by direct intrahippocampal microinjection, accompanied and followed by in vivo microdialysis for metabolites in conscious, behaving animals.

Hipokampal İnsülin Mikroenjeksiyon ve<em&gt; In vivo</emMekansal Bellek Test sırasında&gt; mikrodivaliz

Glucose metabolism is a useful marker for local neural activity, forming the basis of methods such as 2-deoxyglucose and functional magnetic resonance ...

A Single-fly Assay for Foraging Behavior in Drosophila

For many animals, hunger promotes changes in the olfactory system in a manner that facilitates the search for appropriate food sources. In this video article, we describe an automated assay to measure the effect of hunger or satiety on olfactory dependent food search behavior in the adult fruit fly Drosophila melanogaster. In a light-tight box illuminated by red light that is invisible to fruit flies, a camera linked to custom data acquisition software monitors the position of six flies simultaneously. Each fly is confined to walk in individual arenas containing a food odor at the center. The testing arenas rest on a porous floor that functions to prevent odor accumulation. Latency to locate the odor source, a metric that reflects olfactory sensitivity under different physiological states, is determined by software analysis. Here, we discuss the critical mechanics of running this behavioral paradigm and cover specific issues regarding fly loading, odor contamination, assay temperature, data quality, and statistical analysis.

A Single-fly Assay for Foraging Behavior in Drosophila

In this video article, we describe an automated assay to measure the effect of hunger or satiety on olfactory dependent food search behavior in the adult fruit fly Drosophila melanogaster.

Toplayıcılık Davranış için Tek-sinek Deneyi<em&gt; Drosophila</em

For many animals, hunger promotes changes in the olfactory system in a manner that facilitates the search for appropriate food sources. In this video ...

A Simple Behavioral Assay for Testing Visual Function in Xenopus laevis

Measurement of the visual function in the tadpoles of the frog, Xenopus laevis, allows screening for blindness in live animals. The optokinetic response is a vision-based, reflexive behavior that has been observed in all vertebrates tested. Tadpole eyes are small so the tail flip response was used as alternative measure, which requires a trained technician to record the subtle response. We developed an alternative behavior assay based on the fact that tadpoles prefer to swim on the white side of a tank when placed in a tank with both black and white sides. The assay presented here is an inexpensive, simple alternative that creates a response that is easily measured. The setup consists of a tripod, webcam and nested testing tanks, readily available in most Xenopus laboratories. This article includes a movie showing the behavior of tadpoles, before and after severing the optic nerve. In order to test the function of one eye, we also include representative results of a tadpole in which each eye underwent retinal axotomy on consecutive days. Future studies could develop an automated version of this assay for testing the vision of many tadpoles at once.

A Simple Behavioral Assay for Testing Visual Function in Xenopus laevis

Xenopus laevis tadpoles prefer swimming on the white side of a black/white tank. This behavior is guided by their vision. Based on this behavior, we present a simple assay to test the visual function of tadpoles.

Test Görsel Fonksiyonu için Basit Bir Davranış Testi<em&gt; Xenopus laevis</em

Measurement of the visual function in the tadpoles of the frog, Xenopus laevis, allows screening for blindness in live animals. The optokinetic response ...

Live Imaging of Drosophila Larval Neuroblasts

Stem cells divide asymmetrically to generate two progeny cells with unequal fate potential: a self-renewing stem cell and a differentiating cell. Given their relevance to development and disease, understanding the mechanisms that govern asymmetric stem cell division has been a robust area of study. Because they are genetically tractable and undergo successive rounds of cell division about once every hour, the stem cells of the Drosophila central nervous system, or neuroblasts, are indispensable models for the study of stem cell division. About 100 neural stem cells are located near the surface of each of the two larval brain lobes, making this model system particularly useful for live imaging microscopy studies. In this work, we review several approaches widely used to visualize stem cell divisions, and we address the relative advantages and disadvantages of those techniques that employ dissociated versus intact brain tissues. We also detail our simplified protocol used to explant whole brains from third instar larvae for live cell imaging and fixed analysis applications.

Live Imaging of Drosophila Larval Neuroblasts

This protocol details a streamlined method used to conduct live cell imaging in the context of an intact larval brain. Live cell imaging approaches are invaluable for the study of asymmetric neural stem cell divisions as well as other neurogenic and developmental processes, consistently uncovering mechanisms that were previously overlooked.

Canlı Görüntüleme<em&gt; Drosophila</em&gt; Larva nöroblastomun

Stem cells divide asymmetrically to generate two progeny cells with unequal fate potential: a self-renewing stem cell and a differentiating cell. Given ...

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by stressed conspecifics due to an odorant released during stress known as the Drosophila stress odorant (dSO). Through the use of the T-maze apparatus, one can quantify the avoidance of the dSO by responder flies in a very affordable and robust assay. Conditions necessary to obtain a strong performance are presented here. A stressful experience is necessary for the flies to emit dSO, as well as enough emitter flies to cause a robust avoidance response to the presence of dSO. Genetic background, but not their group size, strongly altered the avoidance of the dSO by the responder flies. Canton-S and Elwood display a higher performance in avoiding the dSO than Oregon and Samarkand strains. This behavioral assay will allow identification of mechanisms underlying this social behavior, and the assessment of the influence of genes and environmental conditions on both emission and avoidance of the dSO. Such an assay can be included in batteries of simple diagnostic tests used to identify social deficiencies of mutants or environmental conditions of interest.

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Here, we present a protocol to quantify the avoidance of stressed individuals. This paradigm is powerful yet user-friendly and can be used to assess the influence of genes and environment on one kind of social interaction in Drosophila melanogaster.

Sosyal Kaçınma Niceleme için Basit Testi<em&gt; Drosophila melanogaster</em

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by ...

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense organs provide an excellent model system for the study of mechanosensation. This protocol describes a simple touch behavior in fruit flies which can be used to identify mutations that interfere with mechanosensation. The tactile stimulation of a macrochaete bristle on the thorax of flies elicits a grooming reflex from either the first or third leg. Mutations that interfere with mechanotransduction (such as NOMPC), or with other aspects of the reflex arc, can inhibit the grooming response. A traditional screen of adult behaviors would have missed mutants that have essential roles during development. Instead, this protocol combines the touch screen with mosaic analysis with a repressible cell marker (MARCM) to allow for only limited regions of homozygous mutant cells to be generated and marked by the expression of green fluorescent protein (GFP). By testing MARCM clones for abnormal behavioral responses, it is possible to screen a collection of lethal p-element mutations to search for new genes involved in mechanosensation that would have been missed by more traditional methods.

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

In order to identify novel mutations affecting mechanosensation, we designed an assay that measures the behavioral response to tactile stimulation of fly bristles in mutant clones generated by the MARCM method. The combination of techniques allows for the identification of mechanosensitive mutations that would otherwise be lethal.

Arasında Mechanosensation için bir davranış tahlilinde klonlarda MARCM tabanlı<em&gt; Drosophila melanogaster</em

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense ...

Taste Preference Assay for Adult Drosophila

Olfactory and gustatory perception of the environment is vital for animal survival. The most obvious application of these chemosenses is to be able to distinguish good food sources from potentially dangerous food sources. Gustation requires physical contact with a chemical compound which is able to signal through taste receptors that are expressed on the surface of neurons. In insects, these gustatory neurons can be located across the animal's body allowing taste to play an important role in many different behaviors. Insects typically prefer compounds containing sugars, while compounds that are considered bitter tasting are avoided. Given the basic biological importance of taste, there is intense interest in understanding the molecular mechanisms underlying this sensory modality. We describe an adult Drosophila taste assay which reflects the preference of the animals for a given tastant compound. This assay may be applied to animals of any genetic background to examine the taste preference for a desired soluble compound.

Taste Preference Assay for Adult Drosophila

Taste is an important sensory process which facilitates attraction to beneficial substances and avoidance of toxic substances. This protocol describes a simple ingestion assay for determining Drosophila gustatory preference for a given chemical compound.

Yetişkin için tat Tercihi Testi<em&gt; Drosophila</em

Olfactory and gustatory perception of the environment is vital for animal survival.  The most obvious application of these chemosenses is to be able to ...

Dynamic Quantitative Sensory Testing to Characterize Central Pain Processing

Central facilitation and modulation of incoming nociceptive signals play an important role in the perception of pain. Disruption in central pain processing is present in many chronic pain conditions and can influence responses to specific therapies. Thus, the ability to precisely describe the state of central pain processing has profound clinical significance in both prognosis and prediction. Because it is not practical to record neuronal firings directly in the human spinal cord, surrogate behavior tests become an important tool to assess the state of central pain processing. Dynamic QST is one such test, and can probe both the ascending facilitation and descending modulation of incoming nociceptive signals via TS and CPM, respectively. Due to the large between-individual variability in the sensitivity to noxious signals, standardized TS and CPM tests may not yield any meaningful data in up to 50% of the population due to floor or ceiling effects. We present methodologies to individualize TS and CPM so we can capture these measures in a broader range of individuals than previously possible. We have used these methods successfully in several studies at the lab, and data from one ongoing study will be presented to demonstrate feasibility and potential applications of the methods.

By assessing pain in response to repetitive or different types of standardized stimuli, dynamic quantitative sensory testing (QST) can reveal changes in the central processing of pain. We present methods to optimize and individualize two dynamic QST measures: temporal summation (TS) and conditioned pain modulation (CPM).

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Kullanarak Bileşiklerin Biyoaktivite için eşli Nanoinjection ve Ekran için Elektrofizyoloji Testi Drosophila melanogaster Dev Elyaf Sistemi

Test için Yöntemler Drosophila Davranış

Hipokampal İnsülin Mikroenjeksiyon ve

Toplayıcılık Davranış için Tek-sinek Deneyi

Test Görsel Fonksiyonu için Basit Bir Davranış Testi

Canlı Görüntüleme

Sosyal Kaçınma Niceleme için Basit Testi

Arasında Mechanosensation için bir davranış tahlilinde klonlarda MARCM tabanlı

Yetişkin için tat Tercihi Testi

Dinamik Kantitatif Duyusal Test Merkezi Ağrı İşleme karakterize etmek