Name: extracellularly identifying motor neurons for a muscle motor pool in aplysia californica
Uploaded: 2013-03-25T12:00:00
Description: Watch this Scientific Journal Video about Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica at JoVE.com

Questa ricerca è stata sostenuta da sovvenzioni NIH e NSF NS047073 concessione DMS1010434.

1. Preparazione del piatto di registrazione <ol><li> Durante gli esperimenti dei trasduttori di forza, i gangli buccale, ganglio cerebrale, e la massa buccale sono posti in un piatto tondo Pyrex che è specializzata per gli studi di forza. </li><li> Per indurre ingestive-come modelli negli esperimenti, abbiamo bisogno di applicare la non idrolizzabile carbacolo agonista colinergico al ganglio cerebrale 15. Per evitare il contatto diretto carbacolo sul gangli buccale e massa buccale, camere separati per isol...

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Biol. 80, 93-118 (1979).</li><li>Peters, M., Altrup, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+organization+in+pharynx+of+Helix+pomatia.">Motor organization in pharynx of Helix pomatia.</a> J. Neurophysiol. 52 (3), 389-409 (1984).</li><li>Church, P. J., Cohen, K. P., Scott, M. L., Kirk, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peptidergic+motoneurons+in+the+buccal+ganglia+of+Aplysia+californica:+immunocytochemical,+morphological,+and+physiological+characterizations.">Peptidergic motoneurons in the buccal ganglia of Aplysia californica: immunocytochemical, morphological, and physiological characterizations.</a> J. Comp. Physiol. A. 168 (3), 323-336 (1991).</li><li>Lu, H., Chestek, C. A., Shaw, K. M., Chiel, H. 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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activity+of+multiple+identified+motor+neurons+recorded+intracellularly+during+evoked+feedinglike+motor+programs+in+Aplysia.">Activity of multiple identified motor neurons recorded intracellularly during evoked feedinglike motor programs in Aplysia.</a> J. Neurophys. 72 (4), 1794-1809 (1994).</li><li>McManus, J. M., Lu, H., Chiel, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+In+Vitro+Preparation+for+Eliciting+and+Recording+Feeding+Motor+Programs+with+Physiological+Movements+in+Aplysia+californica.">An In Vitro Preparation for Eliciting and Recording Feeding Motor Programs with Physiological Movements in Aplysia californica.</a> J. Vis. Exp. (70), e4320 (2012).</li><li>Cullins, M. J., Chiel, H. 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F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+and+synaptic+activation+of+the+fast+coxal+depressor+motoneurone+of+the+cockroach.+Periplaneta+americana.">Structure and synaptic activation of the fast coxal depressor motoneurone of the cockroach. Periplaneta americana.</a> J. Exp. Biol. 56 (3), 647-656 (1972).</li><li>Westerfield, M., McMurray, J. V., Eisen, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identified+motoneurons+and+their+innervation+of+axial+muscles+in+the+zebrafish.">Identified motoneurons and their innervation of axial muscles in the zebrafish.</a> J. Neurosci. 6 (8), 2267-2277 (1986).</li><li>Susswein, A. J., Rosen, S. 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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activity+patterns+of+the+B31/B32+pattern+initiators+innervating+the+I2+muscle+of+the+buccal+mass+during+normal+feeding+movements+in+Aplysia+californica.">Activity patterns of the B31/B32 pattern initiators innervating the I2 muscle of the buccal mass during normal feeding movements in Aplysia californica.</a> J. Neurophys. 75 (4), 1309-1326 (1996).</li><li>Morton, D. W., Chiel, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+buccal+nerve+activity+that+distinguishes+ingestion+from+rejection+can+be+used+to+predict+behavioral+transitions+in+Aplysia.">In vivo buccal nerve activity that distinguishes ingestion from rejection can be used to predict behavioral transitions in Aplysia.</a> J. Comp. Physiol. A. 172 (1), 17-32 (1993).</li><li>Warman, E. N., Chiel, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+technique+for+chronic+single-unit+extracellular+recording+in+freely+behaving+animals+using+pipette+electrodes.">A new technique for chronic single-unit extracellular recording in freely behaving animals using pipette electrodes.</a> J. Neurosci. Methods. 57 (2), 161-169 (1995).</li><li>Nargeot, R. N., Baxter, D. A., Byrne, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contingent-dependent+enhancement+of+rhythmic+motor+patterns:+an+in+vitro+analog+of+operant+conditioning.">Contingent-dependent enhancement of rhythmic motor patterns: an in vitro analog of operant conditioning.</a> J. Neurosci. 17 (21), 8093-8105 (1997).</li><li>Kandel, E. 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> Behavioral biology of Aplysia. , (1979).</li><li>Scott, M. L., Govind, C. K., Kirk, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuromuscular+organization+of+the+buccal+system+in+Aplysia+californica.">Neuromuscular organization of the buccal system in Aplysia californica.</a> J. Comp. Neurol. 312 (2), 207-222 (1991).</li><li>Rosen, S. C., Miller, M. W., Cropper, E. C., Kupfermann, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Outputs+of+radula+mechanoafferent+neurons+in+Aplysia+are+modulated+by+motor+neurons,+interneurons,+and+sensory+neurons.">Outputs of radula mechanoafferent neurons in Aplysia are modulated by motor neurons, interneurons, and sensory neurons.</a> J. Neurophysiol. 83 (3), 1621-1636 (2000).</li><li>Rosen, S. C., Miller, M. W., Evans, C. G., Cropper, E. C., Kupfermann, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diverse+synaptic+connections+between+peptidergic+radula+mechanoafferent+neurons+and+neurons+in+the+feeding+system+of+Aplysia.">Diverse synaptic connections between peptidergic radula mechanoafferent neurons and neurons in the feeding system of Aplysia.</a> J. Neurophysiol. 83 (3), 1605-1620 (2000).</li><li>Weiss, K. R., Chiel, H. J., Koch, U., Kupfermann, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activity+of+an+identified+histaminergic+neuron,+and+its+possible+role+in+arousal+of+feeding+behavior+in+semi-intact+Aplysia.">Activity of an identified histaminergic neuron, and its possible role in arousal of feeding behavior in semi-intact Aplysia.</a> J. Neurosci. 6 (8), 2403-2415 (1986).</li><li>Rosen, S. C., Teyke, T., Miller, M. W., Weiss, K. R., Kupfermann, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+characterization+of+cerebral-to-buccal+interneurons+implicated+in+the+control+of+motor+programs+associated+with+feeding+in+Aplysia.">Identification and characterization of cerebral-to-buccal interneurons implicated in the control of motor programs associated with feeding in Aplysia.</a> J. Neurosci. 11 (11), 3630-3655 (1991).</li><li>Jing, J., Weiss, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+variants+of+a+motor+act+in+a+modular+and+hierarchical+motor+network.">Generation of variants of a motor act in a modular and hierarchical motor network.</a> Curr. Biol. 15 (19), 1712-1721 (2005).</li><li>Azizi, F., Lu, H., Chiel, H. J., Mastrangelo, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+neurostimulation+using+pulse+code+modulation+(PCM)+microfluidic+chips.">Chemical neurostimulation using pulse code modulation (PCM) microfluidic chips.</a> J. Neurosci. Methods. 192 (2), 193-198 (2010).</li><li>Zhurov, Y., Proekt, A., Weiss, K. 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Negli animali con grandi neuroni identificati, come molluschi (ad esempio, Lymnaea, Helix, e Aplysia), analisi di piscine motore è in genere fatto utilizzando la registrazione intracellulare 1,2,3,4. In questo protocollo, si descrive un processo per identificare univocamente i neuroni motori per un pool motore utilizzando una tecnica extracellulare. Abbiamo usato le misure di forza, come un esempio di questo processo. Si potrebbe anche usare EMG per misurare innervazioni muscolari. Brevemen...

<table border="1"><tbody><tr><td> Nome </td><td> Azienda </td><td> Numero di catalogo </td><td> Commenti </td></tr><tr><td> Cloruro di sodio </td><td> Fisher Scientific </td><td> S671 </td><td> Biologica, certificata </td></tr><tr><td> Cloruro di potassio </td><td> Fisher Scientific </td><td> P217 </td><td> Certificato ACS </td></tr><tr><td> Magnesio cloruro esaidrato </td><td> Acros Organics </td><td> 19753 </td><td> 99% </td></tr><tr><td> Solfato di magnesio eptaidrato </td><td> Fisher Scientific </td><td> M63 </td><td> Certificato ACS </td></tr><tr><td> Cloruro di calcio diidrato </td><td> Fisher Scientifica </td><td> C79 </td><td> Certificato ACS </td></tr><tr><td> Glucosio (destrosio) </td><td> Sigma-Aldrich </td><td> G7528 </td><td> BioXtra </td></tr><tr><td> MOPS buffer </td><td> Acros Organics </td><td> 17263 </td><td> 99% </td></tr><tr><td> Carbacolo </td><td> Acros Organics </td><td> 10824 </td><td> 99% </td></tr><tr><td> Soda caustica </td><td> Fisher Scientific </td><td> SS255 </td><td> Certificato </td></tr><tr><td> Acido cloridrico </td><td> Fisher Scientific </td><td> SA49 </td><td> Certificato </td></tr><tr><td> Singola canna capillare di vetro </td><td> AM Sistemi </td><td> 6150 </td><td></td></tr><tr><td> Flaming-Brown micropipetta estrattore modello P-80/PC </td><td> Sutter Instruments </td><td></td><td> Incandescenza usata: FT345B </td></tr><tr><td> Smalto filo rivestito in acciaio inox </td><td> California filo sottile </td><td></td><td> 0.001D, rivestimento h </td></tr><tr><td> Delle famiglie in silicone II Colla </td><td> GE </td><td></td><td></td></tr><tr><td> Duro Quick-Gel supercolla </td&gt; <td> Henkel corp. </td><td></td><td></td></tr><tr><td> AM Sistemi amplificatore modello 1700 </td><td> AM Sistemi </td><td></td><td> Le impostazioni del filtro: 10-500 Hz per il nervo I2 / muscolare, 300-500 Hz per tutti gli altri nervi </td></tr><tr><td> Pulsemaster multicanale Stimolatore </td><td> Mondo Strumenti di precisione </td><td> A300 </td><td></td></tr><tr><td> Stimolo Isolator </td><td> Mondo Strumenti di precisione </td><td> A360 </td><td></td></tr><tr><td> AxoGraph X </td><td> AxoGraph scientifico </td><td></td><td> Software per le registrazioni </td></tr><tr><td> Pin del connettore d&#39;oro </td><td> Bulgin </td><td> SA3148 / 1 </td><td></td></tr><tr><td> Gold Connector Sockets </td><td> Bulgin </td><td> SA3149 / 1 </td><td></td></tr><tr><td> Sylgard 184 in silicone elastomero </td><td> Dow Corning </td><td></td><td></td></tr><tr><td> 100 x 15 millimetri piatto cristallizzazione </td><td> Pyrex </td><td></td><td></td></tr><tr><td> Vuoto Grasso Alta </td><td> Dow Corning </td><td></td><td></td></tr><tr><td> Pipetta Suggerimenti </td><td> Fisher Scientific </td><td> 21-375D </td><td></td></tr><tr><td> Minutien Pins </td><td> Strumenti Scienza Belle </td><td> 26002-10 </td><td></td></tr><tr><td> Modellazione argilla </td><td> Sargent Art </td><td> 22-4400 </td><td></td></tr><tr><td> Whisper pompa ad aria </td><td> Tetra </td><td> 77849 </td><td></td></tr><tr><td> Acquario Tubi </td><td> Eheim </td><td> 7783 </td><td> 12/16 mm </td></tr><tr><td> Elite Diffusore </td><td> Hagen </td><td> A962 </td><td></td></tr><tr><td> Vannas Primavera Forbici </td><td> Strumenti Scienza Belle </td><td> 15000-08 </td><td></td></tr><tr><td> Dumont # 5 pinza sottile </td><td> Belle SciencStrumenti elettronici </td><td> 11254-20 </td><td></td></tr><tr><td> Kimwipes </td><td> Kimberly-Clark </td><td> 34155 </td><td></td></tr></tbody></table>

extracellularly identifying motor neurons for a muscle motor pool in aplysia californica

Negli animali con grandi neuroni identificati (ad esempio molluschi), analisi di piscine motore viene fatto utilizzando tecniche intracellulari 1,2,3,4. Recentemente, abbiamo sviluppato una tecnica per stimolare extracellulare e registrare i singoli neuroni in Aplysia californica 5. Descriviamo ora un protocollo per utilizzare questa tecnica per identificare e caratterizzare motoneuroni all&#39;interno di un pool motore. Questa tecnica ha vantaggi extracellulare. Primi elettrodi extracellulari, può stimolare e registrare i neuroni attraverso la guaina 5, quindi non ha bisogno di essere rimosso. Così, i neuroni saranno più sani in esperimenti extracellulari che in quelli intracellulari. Secondo, se gangli sono ruotate di appropriate pinning della guaina, elettrodi extracellulari possono accedere neuroni su entrambi i lati del ganglio, che rende più facile e più efficace per identificare neuroni multipli nella stessa preparazione. In terzo luogo, extracellulareLAR elettrodi non devono penetrare nelle cellule, e quindi può essere facilmente spostato avanti e indietro tra neuroni, causando meno danni. Ciò è particolarmente utile quando si cerca di registrare i neuroni multipli durante ripetere schemi motori che possono persistere per qualche minuto. In quarto luogo, gli elettrodi extracellulari sono più flessibili di quelli intracellulari durante i movimenti muscolari. Elettrodi intracellulari può tirare fuori e danneggiare i neuroni durante le contrazioni muscolari. Al contrario, poiché elettrodi extracellulari pigiate delicatamente sulla guaina sopra neuroni, di solito soggiornare sopra il neurone stesso durante le contrazioni muscolari, e quindi può essere utilizzato in più preparazioni intatti. Per identificare in modo univoco i neuroni motori per un parco macchine (in particolare, la I1/I3 muscolo in Aplysia) con elettrodi extracellulari, si possono usare le funzioni che non richiedono misure intracellulari come criteri: dimensione del soma e la posizione, la proiezione assonale, e innervazione muscolare 4, 6,7. Per la piscina particolare motore utilizzato per illustrare la tecnica, abbiamo registrato da nervi vestibolari 2 e 3 per misurare proiezioni assonali, e misurato le forze di contrazione del muscolo I1/I3 per determinare il modello di innervazione muscolare per i motoneuroni singoli. Dimostriamo il processo completo della prima motoneuroni identificano con innervazione del muscolo, quindi caratterizzare la loro tempistica durante schemi motori, la creazione di un metodo semplificato diagnostico per l&#39;identificazione rapida. Il metodo più semplice e rapida diagnosi è superiore per preparazioni più intatte, ad esempio nella preparazione massa sospesa buccale 8 o 9 in vivo. Questo processo può essere applicato anche in altri autoveicoli piscine 10,11,12 in Aplysia o in altri sistemi animali 2,3,13,14.

Le figure 4 e 5 mostrano i risultati tipici per identificare due I1/I3 motoneuroni. Figura 4 mostra le registrazioni soma di un grande motoneuroni, B3, durante schemi egestive-simili e ingestive-like (figure 4C, 4D). L&#39;uno-per-uno picchi corrispondenti sul canale soma e il canale ipsilaterale BN2 (figura 4E) mostrano che la specificità di registrazione B3 soma è stata mantenuta durante modelli. B3 incendi durante il medio-tardiva ...

Watch this Scientific Journal Video about Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica at JoVE.com

Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica

Negli animali con grandi neuroni identificati (<em&gt; Ad esempio,</em&gt; Molluschi), l&#39;analisi delle piscine motore viene fatto utilizzando tecniche intracellulari<sup&gt; 1,2,3,4</sup&gt;. Recentemente, abbiamo sviluppato una tecnica per stimolare extracellulare e registrare i singoli neuroni in<em&gt; Aplysia californica</em<sup&gt; 5</sup&gt;. Descriviamo ora un protocollo per utilizzare questa tecnica per identificare e caratterizzare motoneuroni all&#39;interno di un pool motore.

Extracellulare Identificazione neuroni motori per un pool di motore muscolare in<em&gt; Aplysia californica</em

In animals with large  identified neurons (e.g. mollusks), analysis of motor pools is done using  intracellular techniques1,2,3,4. Recently,  we developed ...

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JoVE Journal

Neuroscience

Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica

Paired Patch Clamp Recordings from Motor-neuron and Target Skeletal Muscle in Zebrafish

Accoppiato a pinza registrazioni patch da Motor-neurone e muscolo scheletrico bersaglio in Zebrafish

Larval zebrafish represent the first vertebrate model system to allow simultaneous patch clamp recording from a spinal motor-neuron and target muscle.  ...

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Neuroscienze

Live-imaging of PKC Translocation in Sf9 Cells and in Aplysia Sensory Neurons

Live-imaging di traslocazione della PKC in SF9 cellule e nei neuroni sensoriali Aplysia

Protein kinase Cs (PKCs) are serine threonine kinases that play a central role in regulating a wide variety of cellular processes such as cell growth and ...

Quantitative Analysis of Synaptic Vesicle Pool Replenishment in Cultured Cerebellar Granule Neurons using FM Dyes

Analisi quantitativa di Replenishment pool delle vescicole sinaptiche in colture di neuroni granuli cerebellari utilizzando coloranti FM

After neurotransmitter release in central nerve terminals, SVs are rapidly retrieved by endocytosis.  Retrieved SVs are then refilled with ...

An In Vitro Preparation for Eliciting and Recording Feeding Motor Programs with Physiological Movements in Aplysia californica

An In Vitro Preparation for Eliciting and Recording Feeding Motor Programs with Physiological Movements in Aplysia californica

Un<em&gt; In Vitro</emPreparazione&gt; per stimolare e registrare i programmi motori Nutrire con i movimenti fisiologici in<em&gt; Aplysia californica</em

Multifunctionality, the ability of one peripheral  structure to generate multiple, distinct behaviors1, allows animals  to rapidly adapt their behaviors ...

Isolation of Sensory Neurons of Aplysia californica for Patch Clamp Recordings of Glutamatergic Currents

Isolation of Sensory Neurons of Aplysia californica for Patch Clamp Recordings of Glutamatergic Currents

Isolamento dei neuroni sensoriali<em&gt; Aplysia californica</em&gt; Per Bloccare Registrazioni patch di correnti glutamatergico

The marine gastropod mollusk Aplysia californica has a venerable  history as a model of nervous system function, with particular significance in  studies ...

Functional and Morphological Assessment of Diaphragm Innervation by Phrenic Motor Neurons

Funzionale e morfologica Valutazione della membrana innervazione da Phrenic Motor Neurons

This protocol specifically focuses on tools for assessing phrenic motor neuron (PhMN) innervation of the diaphragm at both the electrophysiological and ...

Intramuscular Injections Along the Motor End Plates: A Minimally Invasive Approach to Shuttle Tracers Directly into Motor Neurons

Iniezioni intramuscolari Lungo le piastre Motor finale: un approccio mini-invasivo di navetta Tracers Direttamente nella Motor Neurons

Diseases affecting the integrity of spinal cord motor neurons are amongst the most debilitating neurological conditions. Over the last decades, the ...

Why Quantification Matters: Characterization of Phenotypes at the Drosophila Larval Neuromuscular Junction

Why Quantification Matters: Characterization of Phenotypes at the Drosophila Larval Neuromuscular Junction

Perché Quantificazione Matters: Caratterizzazione dei fenotipi al<em&gt; Drosophila</em&gt; Larvale Giunzione neuromuscolare

Most studies on morphogenesis rely on qualitative descriptions of how anatomical traits are affected by the disruption of specific genes and genetic ...

Retrograde Neuroanatomical Tracing of Phrenic Motor Neurons in Mice

Analisi Neuroanatomical retrogrado dei motoneuroni frenico in topi

Phrenic motor neurons are cervical motor neurons originating from C3 to C6 levels in most mammalian species. Axonal projections converge into phrenic ...

A Minimally Invasive Lesion Technique for Muscles Intrinsic to the Odontophore of Aplysia californica

A Minimally Invasive Lesion Technique for Muscles Intrinsic to the Odontophore of Aplysia californica

Una tecnica di lesione mini-invasiva per i muscoli intrinseca all'odontoia di Aplysia californica

Aplysia californica is a model system for studying the neural control of learning and behavior. This animal has a semi-open circulatory system, making it ...