Name: a molecular readout of long-term olfactory adaptation in c. elegans
Uploaded: 2012-12-22T14:00:00
Description: Watch this Scientific Journal Video about A Molecular Readout of Long-term Olfactory Adaptation in C. elegans at JoVE.com

हम इस पांडुलिपि के सावधान पढ़ने के लिए स्कॉट हैमिल्टन, और O&#39;Halloran प्रयोगशाला के सदस्यों का शुक्रिया अदा करना चाहते हैं. हम भी हमारे गुमनाम समीक्षक उत्कृष्ट सुझाव और insightful टिप्पणियों के लिए धन्यवाद.

1. GFP का निर्माण EGL-4 व्यक्त पशु Tagged <ol><li> जीन ओडीआर 3 (2678 बीपी सीधे शुरू कोडोन के अपस्ट्रीम उपयोग करें) के लिए प्रमोटर के तहत translational संलयन :: GFP क्लोन EGL-4: (पी) ओडीआर-3 :: GFP :: EGL-4. अभिव्यक्ति amphid न्यूरॉन जोड़े में ओडीआ?...

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	<li>Colbert, H. A., Bargmann, C. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Odorant-specific+adaptation+pathways+generate+olfactory+plasticity+in+C.+elegans.">Odorant-specific adaptation pathways generate olfactory plasticity in C. elegans.</a> Neuron. 14 (4), 803 (1995).</li><li>Colbert, H. A., Bargmann, C. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Environmental+signals+modulate+olfactory+acuity,+discrimination,+and+memory+in+Caenorhabditis+elegans.">Environmental signals modulate olfactory acuity, discrimination, and memory in Caenorhabditis elegans.</a> Learning and Memory. 4 (2), 179 (1997).</li><li>Bargmann, C. 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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=Efficient+gene+transfer+in+C.elegans:+extrachromosomal+maintenance+and+integration+of+transforming+sequences.">Efficient gene transfer in C.elegans: extrachromosomal maintenance and integration of transforming sequences.</a> EMBO Journal. 10, 3959-39 (1991).</li><li>Miyabayashi, 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=Expression+and+function+of+members+of+a+divergent+nuclear+receptor+family+in+Caenorhabditis+elegans.">Expression and function of members of a divergent nuclear receptor family in Caenorhabditis elegans.</a> Dev. Biol. 215, 314 (1999).</li><li>L'Etoile, N. D., Bargmann, C. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfaction+and+odor+discrimination+are+mediated+by+the+C.+elegans+guanylyl+cyclase+ODR-1.">Olfaction and odor discrimination are mediated by the C. elegans guanylyl cyclase ODR-1.</a> Neuron. 25 (3), 575 (2000).</li><li>Mello, C., Fire, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DNA+transformation.">DNA transformation.</a> Methods Cell Biol. 48, 451 (1995).</li><li>Stiernagle, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maintenance+of+C.+elegans.">Maintenance of C. elegans.</a> Wormbook. , (2006).</li><li>Brenner, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetics+of+Caenorhabditis+elegans.">The genetics of Caenorhabditis elegans.</a> Genetics. 77 (1), 71 (1974).</li><li>Kauffman, A., Parsons, L., Stein, G., Wills, A., Kaletsky, R., Murphy, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=C.+elegans+Positive+Butanone+Learning,+Short-term,+and+Long-term+Associative+Memory+Assays.">C. elegans Positive Butanone Learning, Short-term, and Long-term Associative Memory Assays.</a> J. 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गंध प्रेरित एक GFP टैग EGL चार अणु परमाणु प्रविष्टि यहाँ वर्णित सी. में घ्राण अनुकूलन का एक मजबूत आणविक readout प्रदान करता है एलिगेंस. गंध से प्रेरित परमाणु स्थानान्तरण assays सीधा कर रहे हैं और तैयारी के सम?...

<table border="1"><tbody><tr><td> अभिकर्मक के नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td><td> टिप्पणियां </td></tr><tr><td> Bacto अग्रवाल </td><td> DIFCO </td><td> DF0140-07-4 </td><td> एन जी एम प्लेटों </td></tr><tr><td> सोडियम क्लोराइड </td><td> फिशर रासायनिक </td><td> S671-10 </td><td> एन जी एम प्लेटों </td></tr><tr><td> Bacto peptone </td><td> DIFCO </td><td> DF0118-07-2 </td><td> एन जी एम प्लेटों </td></tr><tr><td> पोटेशियम फॉस्फेट Dibasic </td><td> फिशर रासायनिक </td><td> S375-500 </td><td> एस बेसल बफर और प्लेट एन जी एम </td></tr><tr><td> पोटेशियम फॉस्फेट अकेले आधार का </td><td> फिशर रासायनिक </td><td> P285-500 </td><td> एस बेसल बफर और प्लेट एन जी एम </td></tr><tr><td> छोटे Kimwipes </td><td> Kimberly-क्लार्क </td><td> LS2770 </td><td></td></tr><tr><td> 100% इथेनॉल </td><td> गोल्ड शील्ड रासायनिककं </td><td> 43196-115 </td><td> chemotaxis assays के लिए गिराए odors </td></tr><tr><td> कैल्सियम क्लोराइड </td><td> सिग्मा Aldrich </td><td> C8106-500G </td><td> एन जी एम प्लेटों </td></tr><tr><td> मैग्नीशियम सल्फेट </td><td> सांसद BIOMEDICALS </td><td> +१,५०,१३६-500G </td><td> एन जी एम प्लेटों </td></tr><tr><td> सोडियम azide 99% </td><td> फिशर साइंटिफिक </td><td> ICN10289180 </td><td> संवेदनाहारी </td></tr><tr><td> Agarose - Ultrapure </td><td> Invitrogen </td><td> 16500-500 </td><td> Agarose पैड </td></tr><tr><td> Benzaldehyde </td><td> सिग्मा Aldrich </td><td> B1334-100g </td><td> AWC गंध </td></tr><tr><td> Butanone, ACS ग्रेड </td><td> सिग्मा Aldrich </td><td> 360,473 - 500ml </td><td> AWC गंध </td></tr><tr><td> Microcentrifuge ट्यूबों - 1.5 मिलीलीटर रंगीन </td><td> Denville </td><td> LS8147 </td><td></td></tr><tr><td> पाश्चर pipet डिस्पोजेबल ग्लास 5-3/4 &quot; </td><td> फिशर साइंटिफिक </td&gt; <td> 13-678-20B </td><td></td></tr><tr><td> Stratalinker </td><td> Stratagene </td><td> 2400 Stratalinker </td><td> यूवी एकीकरण </td></tr><tr><td> फ़िल्टर वैक्यूम बोतल - 500 मिलीलीटर </td><td> Nalgene </td><td> 09-740-25B </td><td></td></tr></tbody></table>

a molecular readout of long-term olfactory adaptation in c. elegans

During sustained stimulation most sensory neurons will adapt their response by decreasing their sensitivity to the signal. The adaptation response helps shape attention and also protects cells from over-stimulation. Adaptation within the olfactory circuit of C. elegans was first described by Colbert and Bargmann1,2. Here, the authors defined parameters of the olfactory adaptation paradigm, which they used to design a genetic screen to isolate mutants defective in their ability to adapt to volatile odors sensed by the Amphid Wing cells type C (AWC) sensory neurons. When wildtype C. elegans animals are exposed to an attractive AWC-sensed odor3 for 30 min they will adapt their responsiveness to the odor and will then ignore the adapting odor in a chemotaxis behavioral assay for ~1 hr. When wildtype C. elegans animals are exposed to an attractive AWC-sensed odor for ~1 hr they will then ignore the adapting odor in a chemotaxis behavioral assay for ~3 hr. These two phases of olfactory adaptation in C. elegans were described as short-term olfactory adaptation (induced after 30 min odor exposure), and long-term olfactory adaptation (induced after 60 min odor exposure). Later work from L'Etoile et al.,4 uncovered a Protein Kinase G (PKG) called EGL-4 that is required for both the short-term and long-term olfactory adaptation in AWC neurons. The EGL-4 protein contains a nuclear localization sequence that is necessary for long-term olfactory adaptation responses but dispensable for short-term olfactory adaptation responses in the AWC4. By tagging EGL-4 with a green fluorescent protein, it was possible to visualize the localization of EGL-4 in the AWC during prolonged odor exposure. Using this fully functional GFP-tagged EGL-4 (GFP::EGL-4) molecule we have been able to develop a molecular readout of long-term olfactory adaptation in the AWC5. Using this molecular readout of olfactory adaptation we have been able to perform both forward and reverse genetic screens to identify mutant animals that exhibit defective subcellular localization patterns of GFP::EGL-4 in the AWC6,7. Here we describe: 1) the construction of GFP::EGL-4 expressing animals; 2) the protocol for cultivation of animals for long-term odor-induced nuclear translocation assays; and 3) the scoring of the long-term odor-induced nuclear translocation event and recovery (re-sensitization) from the nuclear GFP::EGL-4 state.

:: GFP EGL 4in स्थानीयकरण AWC पहले पैटर्न और लंबे समय तक गंध के प्रदर्शन के बाद के एक उदाहरण चित्रा 2 में दिखाया गया है. पिछले लंबे समय तक गंध जोखिम, GFP :: EGL-4 AWC (चित्रा 2B) की cytosol स्थानीय है, और के बाद 80 मिनट में ग...

Watch this Scientific Journal Video about A Molecular Readout of Long-term Olfactory Adaptation in C. elegans at JoVE.com

A Molecular Readout of Long-term Olfactory Adaptation in C. elegans

यहाँ हम लंबी अवधि में घ्राण अनुकूलन के एक आणविक readout का वर्णन<em&gt; Caenorhabditis एलिगेंस</em&gt;. प्रोटीन kinase जी, EGL-4, प्राथमिक संवेदी न्यूरॉन AWC बुलाया जोड़ी में स्थिर अनुकूलन प्रतिक्रियाओं के लिए आवश्यक है. लंबे समय तक गंध जोखिम EGL 4 cytosol से AWC के नाभिक को translocates के दौरान.

लंबी अवधि में घ्राण अनुकूलन के एक आणविक Readout<em&gt; सी. एलिगेंस</em

During sustained stimulation most sensory neurons will adapt their response by decreasing their sensitivity to the signal. The adaptation response helps ...

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A Molecular Readout of Long-term Olfactory Adaptation in C. elegans

C. elegans Positive Butanone Learning, Short-term, and Long-term Associative Memory Assays

C. elegans Positive Butanone Learning, Short-term, and Long-term Associative Memory Assays

सी एलिगेंस सकारात्मक Butanone लर्निंग, लघु अवधि, और लंबी अवधि के साहचर्य मेमोरी Assays

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में Synaptic प्रतिदीप्ति स्वचालित में मात्रा<em&gt; सी. एलिगेंस</em

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माउस Cochlear explants की लंबी अवधि के समय चूक इमेजिंग

Here we present a method for long-term time-lapse imaging of live embryonic mouse cochlear explants. The developmental program responsible for building ...

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की स्वचालित विश्लेषण<em&gt; सी। एलिगेंस</em&gt; तैरना व्यवहार Celest सॉफ्टवेयर का उपयोग

Dissecting the neuronal and neuromuscular circuits that regulate behavior remains a major challenge in biology. The nematode Caenorhabditis elegans has ...

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पुरुष C. एलिगेंस सिर ंयूरॉंस में Pheromones के जवाब में ंयूरॉन गतिविधि के इमेजिंग के लिए एक अनुकूलित Microfluidic घ्राण चिप का प्रयोग

The use of calcium indicators has greatly enhanced our understanding of neural dynamics and regulation. The nematode Caenorhabditis elegans, with its ...