Name: paired nanoinjection and electrophysiology assay to screen for bioactivity of compounds using the drosophila melanogaster giant fiber system
Uploaded: 2012-04-15T14:00:00
Description: Watch this Scientific Journal Video about Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System at JoVE.com

हम विशेष Aline Yonezawa में मारी प्रयोगशाला और Godenschwege के प्रयोगशाला, के सदस्यों को स्वीकार करना और टिप्पणी के लिए, इस प्रोटोकॉल के साथ मदद की. इस काम के मस्तिष्क संबंधी विकार के लिए राष्ट्रीय संस्थान और एफएम और टैग करने के लिए स्ट्रोक R21NS06637 अनुदान द्वारा वित्त पोषित किया गया था, अटल बिहारी राष्ट्रीय विज्ञान फाउंडेशन पुरस्कार 082,925 संख्या, URM द्वारा वित्त पोषित किया गया था: भविष्य के शोधकर्ताओं के लिए एकीकृत जीवविज्ञान.

1. इलैक्ट्रोफिजियोलॉजी रिग सेट अप <ol><li> इलेक्ट्रोफिजियोलॉजी रिग सेट अप के लिए आवश्यक उपकरणों के विस्तार में Augustin एट अल द्वारा वर्णित है. इस जर्नल में 14. Electrophysiological जरूरत apparatuses के एक विस्तृत विवरण के लिए ...

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J., Partridge, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+Recordings+from+the+Giant+Fiber+Pathway+of+D.+melanogaster.">Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster.</a> J. Vis. Exp. (47), e2412 (2011).</li><li>Allen, M. J., Godenschwege, T., Zhang, B., Freeman, M. R., Waddell, S. <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 Neurobiology. , 215-224 (2010).</li><li>Blagburn, J. M., Alexopoulos, H., Davies, J. A., Bacon, J. 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यहाँ प्रस्तुत / nanoinjection इलेक्ट्रोफिजियोलॉजी bioassay फल मक्खी के तंत्रिका तंत्र में यौगिकों की एक तेजी से जांच के लिए अनुमति देता है. यह vivo में तकनीक है कि परिसर के छोटे मात्रा की आवश्यकता के लिए एक अच्छी तरह...

<table border="1"><tbody><tr><td> अभिकर्मक का नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td><td> टिप्पणियाँ </td></tr><tr><td> रिकॉर्डिंग कांच इलेक्ट्रोड: borosilicate ग्लास के capillaries </td><td> दुनिया प्रेसिजन उपकरण </td><td> 1B100F - 4 </td><td> 1.0mm आयुध डिपो, 0.58mm आईडी </td></tr><tr><td> उत्तेजक औधधि </td><td> घास उपकरण </td><td> S48 मॉडल </td><td></td></tr><tr><td> प्रवर्धक </td><td> उपकरण इंक हो रही है </td><td> मॉडल 5A </td><td></td></tr><tr><td> डाटा अधिग्रहण सॉफ्टवेयर: Digidata के के </td><td> आणविक डिवाइसेज </td><td> मॉडल 1440A </td><td></td></tr><tr><td> डेटा संग्रह सॉफ्टवेयर: pCLAMP </td><td> आणविक डिवाइसेज </td><td> संस्करण 10 </td><td></td></tr><tr><td> Stereomicroscope डब्ल्यूith फाइबर ऑप्टिक खुर्दबीन अंगूठी प्रकाशक </td><td> AmScope </td><td> एस.एम. 4T HL250-ए.आर. मॉडल </td><td></td></tr><tr><td> बढ़ते के लिए गुंजाइश विदारक </td><td> AmScope </td><td> एस.एम. 2TZ </td><td></td></tr><tr><td> यह पतंग मैनुअल micromanipulator और बेस झुकाना </td><td> दुनिया प्रेसिजन उपकरण </td><td> M3301 # मॉडल पतंग: मॉडल # पतंग - एम 3-L </td><td></td></tr><tr><td> ड्रोसोफिला मेलानोगास्टर जंगली 10E जीनोटाइप (जंगली प्रकार तनाव) </td><td> ब्लूमिंगटन स्टॉक केंद्र </td><td> 3892 # स्टॉक </td><td></td></tr><tr><td> ऊर्ध्वाधर विंदुक डांड़ी </td><td> दाऊद KOPF उपकरण </td><td> 700C मॉडल </td><td></td></tr><tr><td> इंजेक्शन गिलास micropipettes: Borosilicate कांच के capillaries </td><td> दुनिया प्रेसिजन उपकरण </td><td> 4878 # सूचीपत्र </tघ&gt; <td> 1.14mm आयुध डिपो, 0.5mm आईडी </td></tr><tr><td> सिलिकॉन तेल </td><td> मछुआ </td><td> # S159-500 सूचीपत्र </td><td></td></tr><tr><td> Beveler </td><td> Sutter उपकरण कं </td><td> के.टी. ब्राउन प्रकार # BV-10 मॉडल </td><td></td></tr><tr><td> Nanoliter2000 </td><td> दुनिया प्रेसिजन उपकरण </td><td> B203XVY # सूचीपत्र </td><td></td></tr><tr><td> ब्लू रंग भोजन </td><td> McCormick </td><td> N / A </td><td> जल, Propylene Glycol, एफडी और सी 1 ब्लू, और 0.1% Propylparaben (परिरक्षक) सामग्री:. </td></tr><tr><td> Methyllycaconitine साइट्रेट (विधायक) </td><td> बायोसाइंस tocris </td><td> 1029 # सूचीपत्र </td><td></td></tr><tr><td> प्लास्टिक मोम लाठी </td><td> स्वास्थ्यकारी निगम (Akron, ओहियो संयुक्त राज्य अमेरिका) </td><td></td><td></td></tr></tboवि&gt; </table>

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.

Watch this Scientific Journal Video about Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System at JoVE.com

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

एक तेजी से<em&gt; Vivo में</em&gt; Neuromodulatory विशालकाय फाइबर प्रणाली (GFS) का उपयोग यौगिकों के लिए परीक्षण परख<em&gt; ड्रोसोफिला मेलानोगास्टर</em&gt; वर्णित है. पशु के सिर में GFS का electrophysiological रिकॉर्डिंग के साथ nanoinjections न्यूरॉन्स या मांसपेशियों पर यौगिकों की bioactivity प्रकट कर सकते हैं.

बनती है और का उपयोग यौगिकों की bioactivity के लिए इलैक्ट्रोफिजियोलॉजी स्क्रीन करने के लिए परख Nanoinjection ड्रोसोफिला मेलानोगास्टर विशालकाय फाइबर सिस्टम

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 ...

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

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

विशालकाय फाइबर के मार्ग से electrophysiological रिकॉर्डिंग डी. मेलानोगास्टर</em

When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" ...

Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection

Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection

वयस्क के पूरा पर्वत तैयार ड्रोसोफिला Ventral तंत्रिका गर्भनाल विशालकाय फाइबर डाई के इंजेक्शन के लिए

To analyze the axonal and dendritic morphology of neurons, it is essential to obtain accurate labeling of neuronal structures. Preparing well labeled ...

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

केन्द्रीय तंत्रिका तंत्र में एकल कक्षों के लेबल<em&gt; ड्रोसोफिला मेलानोगास्टर</em

In this  article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the ...

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

में सहज हरकत गतिविधि का निर्धारण<em&gt; ड्रोसोफिला मेलानोगास्टर</em

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such ...

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

सामाजिक परिहार की मात्रा में लिए सीधा परख<em&gt; ड्रोसोफिला मेलानोगास्टर</em

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

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

गंध निर्देशित व्यवहार के उच्च संकल्प मात्रा<em&gt; ड्रोसोफिला मेलानोगास्टर</em&gt; का प्रयोग<em&gt; Flywalk</em&gt; प्रतिमान

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

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

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

के Mechanosensation के लिए एक व्यवहार परख क्लोन में MARCM आधारित<em&gt; ड्रोसोफिला मेलानोगास्टर</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 ...

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

उच्च throughput परख की जांच करने के लिए व्यक्ति की अंडे बिछाने प्राथमिकताएं<em&gt; ड्रोसोफिला मेलानोगास्टर</em

Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. ...

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

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

इनाम व्यवहार की मांग में बदलाव को मापने के लिए उपयोग कर एक सरल तरीका<em&gt; ड्रोसोफिला मेलानोगास्टर</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 CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

केशिका फीडर परख में खाद्य सेवन के उपाय<em&gt; ड्रोसोफिला मेलानोगास्टर</em

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of ...