Name: simultaneous intracellular recording of a lumbar motoneuron and the force produced by its motor unit in the adult mouse in vivo
Uploaded: 2012-12-05T12:00:00
Description: Watch this Scientific Journal Video about Simultaneous Intracellular Recording of a Lumbar Motoneuron and the Force Produced by its Motor Unit in the Adult Mouse In vivo at JoVE.com

Dette arbeidet ble gjort mulig takket være økonomisk støtte fra Fondation pour la Recherche Medicale (FRM), Milton Safenowitz postdoktorstilling for ALS Research (ALS Association), NIH NINDS Grants NS05462 og NS034382, og ANR Grant HyperMND.

1. Step One Pre-anestesi medisiner: 10-15 min før induksjon av anestesi, injiserer atropin (0,20 mg / kg) og methylprenidsolone (0,05 mg) subkutant unngå spytt og ødem, henholdsvis. 2. Trinn to Induksjon av anestesi: injisere pentobarbital natrium (70 mg / kg) eller en blanding av ketamin / xylazin (100 mg / kg og 10 mg / kg, henholdsvis) intra-peritoneally. La musen gå under før noen tå klype refleks kan skaffes. Hvis anestesi virk...

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Fremstillingen er beskrevet her er den første som tillater, i den voksne mus, simultan intracellulær opptak av en lumbal motoneuron og måling av den kraft som produseres av muskelfibrene innervated av axon sin. På grunn av den lille størrelsen på dyret, kan de kirurgiske ferdigheter som kreves for dette preparatet være utfordrende å tilegne seg. Imidlertid, når disse ferdigheter er mestret, kan hele operasjonen utføres i tre timer, og dyrene kan overleve i opptil 7 flere timer ette...

A correction was made to <a href="http://www.jove.com/video/4312">Simultaneous Intracellular Recording of a Lumbar Motoneuron and the Force Produced by its Motor Unit in the Adult Mouse In vivo</a>. There was an error in the name of one author, Marin Manuel. The author's name has been corrected to:
Marin Manuel 
 
instead of: 
 
Manuel Marin

A correction to author list has been made for article <a href="http://www.jove.com/video/4312">Simultaneous Intracellular Recording of a Lumbar Motoneuron and the Force Produced by its Motor Unit in the Adult Mouse In vivo</a>.

Erratum: Simultaneous Intracellular Recording of a Lumbar Motoneuron and the Force Produced by its Motor Unit in the Adult Mouse In Vivo

<table border="1"><tbody><tr><td> Navnet på reagensen </td><td> Selskapet </td><td> Katalognummer </td><td> Kommentarer (valgfritt) </td></tr><tr><td> Atropin sulfat </td><td> Aguettant </td><td></td><td></td></tr><tr><td> Methylprenidsolone </td><td> Pfizer </td><td> Solu-Medrol </td><td></td></tr><tr><td> Sodium pentobarbitone </td><td> Sanofi-Aventis </td><td> Pentobarbital </td><td></td></tr><tr><td> Ketamin </td><td></td><td></td><td></td></tr><tr><td> Xylazin </td><td></td><td></td><td></td></tr><tr><td> Glukose </td><td></td><td></td><td></td></tr><tr><td> Plasma expander </td><td> Roger Bellon </td><td> Plasmagel </td><td></td></tr><tr><td> Sløv saks </td><td> FST </td><td> 14079-10 </td><td></td></tr><tr><td> Blunt fin saks </td><td> FST </td><td> 15025-10 </td><td></td></tr><tr><td> Vännäs Spring saks </td><td> FST </td><td> 15002-08 </td><td></td></tr><tr><td> Fin tang taggete </td><td> FST </td><td> 11370-32 </td><td></td></tr><tr><td> Fin tang taggete </td><td> FST </td><td> 11370-31 </td><td></td></tr><tr><td> Cunningham Spinal Adaptor </td><td> Stoelting Co </td><td></td><td></td></tr><tr><td> Kwik-Cast tetningsmasse </td><td> WPI </td><td> # KWIK-CAST </td><td></td></tr><tr><td> Ventilator </td><td> CWE Inc </td><td> SAR-830/AP </td><td></td></tr><tr><td> Kapnografen </td><td> CWE Inc </td><td> μcapstar </td><td></td></tr><tr><td> Varmeteppet </td><td> Harvard Apparatus </td><td> 507221F </td><td></td></tr><tr><td> Intracellulær forsterker </td><td> Axon instruments </td><td> Axoclamp 2B </td><td></td></tr><tr><td> Pipette avtrekker </td><td> Sutter Instruments </td><td> P-97 </td><td></td></tr><tr><td> KCl </td><td> Sigma-Aldrich </td><td> P9333-500G </td><td></td></tr></tbody></table>

simultaneous intracellular recording of a lumbar motoneuron and the force produced by its motor unit in the adult mouse in vivo

Spinal motoneuron har lenge vært en god modell system for å studere nervefunksjoner fordi det er et nevron av det sentrale nervesystemet med de unike egenskapene (1) som har lett identifiserbare mål (muskelfibrene) og derfor har en svært velkjent funksjon (for å kontrollere muskelkontraksjon), (2) å være den konvergent målet for mange rygg-og synkende nettverk, derav navnet &quot;endelige felles vei&quot;, og (3) har et stort soma som gjør det mulig å trenge dem med skarpe intracellulære elektroder . Videre, når undersøkt in vivo, er det mulig å registrere samtidig den elektriske aktiviteten i motoneurons og kraften utviklet av deres muskel mål. Utføre intracellulære opptak av motoneurons in vivo derfor sette experimentalist i den unike posisjon av å kunne studere, på samme tid, alle avdelinger av &quot;motorenhet&quot; (navnet gitt til motoneuron, dens axon, ogmuskelfibrene det innervates 1): innganger impinging på motoneuron de elektrofysiologiske egenskaper motoneuron, og virkningen av disse egenskapene på den fysiologiske funksjon av motoneurons, dvs. kraft produsert av sin motorenheten. Imidlertid er denne tilnærmingen svært utfordrende fordi fremstillingen kan ikke være lammet og dermed den mekaniske stabiliteten for den intracellulære opptaket er redusert. Dermed har denne typen eksperimenter kun oppnådd i katter og i rotter. Imidlertid kunne studere spinal motor systemer gjør en formidabel sprang hvis det var mulig å utføre lignende eksperimenter i normal og genmodifiserte mus. Av tekniske årsaker har studien av spinal nettverk i mus stort sett blitt begrenset til neonatal in vitro preparater, der motoneurons og spinal nettverk er umoden, er motoneurons skilt fra sine mål, og når studert i skiver, den motoneurons er atskilt fra de fleste av sine innganger. Inntil nylig hadde bare noen få grupper klarte å utføre intracellulære opptak av motoneurons in vivo 2-4, herunder vårt team som publiserte et nytt preparat som tillater oss å få svært stabile opptak av motoneurons in vivo hos voksne mus 5,6. Imidlertid ble disse innspillingene oppnådd i lamme dyr, dvs. uten muligheten til å spille kraften produksjon av disse motoneurons. Her presenterer vi en forlengelse av denne originalpreparatet der vi var i stand til å skaffe samtidige opptak av elektrofysiologiske egenskapene til motoneurons og av kraften utviklet av deres motorenheten. Dette er et viktig resultat, som det tillater oss å identifisere de ulike typer motoneurons basert på deres styrke profil, og dermed avsløre deres funksjon. Sammen med genetiske modeller urovekkende spinal segmentell krets 7-9, eller reproducting human diseaSE 10,11, forventer vi denne teknikken til å være et viktig verktøy for å studere spinal motor system.

Figur 1 viser hvordan å identifisere en motoneuron fra Triceps surae gruppen etter penetrasjon. Ved lav stimulering intensitet, kan bare en monosynaptisk EPSP overholdes (Figur 1A). Ved høyere intensitet, kan EPSP være store nok til å utløse en &quot;orthodromic&quot; spike (figur 1B). På enda høyere stimulering intensitet, vises en alt-eller-ingenting antidromic pigg, med en kortere ventetid enn monosynaptisk EPSP (figur 1C). Hvis nok strøm inj...

Watch this Scientific Journal Video about Simultaneous Intracellular Recording of a Lumbar Motoneuron and the Force Produced by its Motor Unit in the Adult Mouse In vivo at JoVE.com

Simultaneous Intracellular Recording of a Lumbar Motoneuron and the Force Produced by its Motor Unit in the Adult Mouse In vivo

Denne nye metoden åpner for samtidig intracellulære opptaket av en enkelt voksen mus motoneuron og måling av den kraft som produseres av sine muskelfibre. Kombinerte undersøkelse av de elektriske og mekaniske egenskaper av motoriske enheter i normal og genmodifiserte dyr er et gjennombrudd for studiet av nevromuskulære system.

Samtidig Intracellulær Innspillingen av en Lumbar Motoneuron og Force Produsert av sin Motor Unit i Adult Mouse<em&gt; In vivo</em

The  spinal motoneuron has long been a good model system for studying neural  function because it is a neuron of the central nervous system with the ...

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Simultaneous Intracellular Recording of a Lumbar Motoneuron and the Force Produced by its Motor Unit in the Adult Mouse In vivo

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