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 arbejde blev gjort muligt takket være økonomisk støtte fra Fondation pour la Recherche MEDICALE (FRM), Milton Safenowitz Postdoc Fellowship for ALS Research (ALS Association), NIH NINDS Grants NS05462 og NS034382, og ANR Grant HyperMND.

1. Trin One Præ-anæstetisk medicinering: 10-15 min før induktion af anæstesi, atropin (0,20 mg / kg) og methylprenidsolone (0,05 mg) subkutant injektion for at forhindre savlen og ødem, hhv. 2. Trin to Induktion af anæstesi: injicere pentobarbitalnatrium (70 mg / kg) eller en blanding af ketamin / xylazin (100 mg / kg og 10 mg / kg) intraperitonealt. Lad musen går under, før nogen toe pinch refleks kan opnås. Hvis anæstesi virke...

<ol>
	<li>Liddel, E. G. T., Sherrington, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recruitment+and+some+other+factors+of+reflex+inhibition.">Recruitment and some other factors of reflex inhibition.</a> Proc. R. Soc. London. B, 488-518 (1925).</li><li>Huizar, P., Kuno, M., Miyata, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+properties+of+spinal+motoneurones+of+normal+and+dystrophic+mice.">Electrophysiological properties of spinal motoneurones of normal and dystrophic mice.</a> The Journal of physiology. 248, 231-246 (1975).</li><li>Alstermark, B., Ogawa, 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+recordings+of+bulbospinal+excitation+in+adult+mouse+forelimb+motoneurons.">In vivo recordings of bulbospinal excitation in adult mouse forelimb motoneurons.</a> Journal of neurophysiology. 92, 1958-1962 (2004).</li><li>Meehan, C. F., Sukiasyan, N., Zhang, M., Nielsen, J. B., Hultborn, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrinsic+properties+of+mouse+lumbar+motoneurons+revealed+by+intracellular+recording+in+vivo.">Intrinsic properties of mouse lumbar motoneurons revealed by intracellular recording in vivo.</a> Journal of neurophysiology. 103, 2599-2610 (2010).</li><li>Manuel, 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=Fast+kinetics,+high-frequency+oscillations,+and+subprimary+firing+range+in+adult+mouse+spinal+motoneurons.">Fast kinetics, high-frequency oscillations, and subprimary firing range in adult mouse spinal motoneurons.</a> J. Neurosci. 29, 11246-11256 (2009).</li><li>Iglesias, 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=Mixed+mode+oscillations+in+mouse+spinal+motoneurons+arise+from+a+low+excitability+state.">Mixed mode oscillations in mouse spinal motoneurons arise from a low excitability state.</a> The Journal of neuroscience: the official journal of the Society for Neuroscience. 31, 5829-5840 (2011).</li><li>Crone, S. A., Zhong, G., Harris-Warrick, R., Sharma, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+mice+lacking+V2a+interneurons,+gait+depends+on+speed+of+locomotion.">In mice lacking V2a interneurons, gait depends on speed of locomotion.</a> J. Neurosci. 29, 7098-7109 (2009).</li><li>Talpalar, A. E., 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=Identification+of+minimal+neuronal+networks+involved+in+flexor-extensor+alternation+in+the+mammalian+spinal+cord.">Identification of minimal neuronal networks involved in flexor-extensor alternation in the mammalian spinal cord.</a> Neuron. 71, 1071-1084 (2011).</li><li>Rabe, N., Gezelius, H., Vallstedt, A., Memic, F., Kullander, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Netrin-1-dependent+spinal+interneuron+subtypes+are+required+for+the+formation+of+left-right+alternating+locomotor+circuitry.">Netrin-1-dependent spinal interneuron subtypes are required for the formation of left-right alternating locomotor circuitry.</a> J. Neurosci. 29, 15642-15649 (2009).</li><li>Gurney, M. E., 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=Motor+neuron+degeneration+in+mice+that+express+a+human+Cu,Zn+superoxide+dismutase+mutation.">Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation.</a> Science. 264, 1772-1775 (1994).</li><li>Cifuentes-Diaz, 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=Neurofilament+accumulation+at+the+motor+endplate+and+lack+of+axonal+sprouting+in+a+spinal+muscular+atrophy+mouse.">Neurofilament accumulation at the motor endplate and lack of axonal sprouting in a spinal muscular atrophy mouse.</a> Hum. Mol. Genet. 11, 1439-1447 (2002).</li><li>Simpson, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prolonged+(12+hours)+intravenous+anesthesia+in+the+rat.">Prolonged (12 hours) intravenous anesthesia in the rat.</a> Laboratory animal science. 47, 519-523 (1997).</li><li>Burke, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+Unit+Types+-+Functional+Specializations+in+Motor+Control.">Motor Unit Types - Functional Specializations in Motor Control.</a> Trends Neurosci. 3, 255-258 (1980).</li><li>Kerkut, G. A., Bagust, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+isolated+mammalian+spinal+cord.">The isolated mammalian spinal cord.</a> Prog. Neurobiol. 46, 1-48 (1995).</li><li>Carp, J. S., 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+in+vitro+protocol+for+recording+from+spinal+motoneurons+of+adult+rats.">An in vitro protocol for recording from spinal motoneurons of adult rats.</a> Journal of Neurophysiology. 100, 474-481 (2008).</li><li>Mitra, P., Brownstone, R. M. <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+Spinal+Cord+Slice+Preparation+for+Recording+from+Lumbar+Motoneurons+of+the+Adult+Mouse.">An In Vitro Spinal Cord Slice Preparation for Recording from Lumbar Motoneurons of the Adult Mouse.</a> Journal of Neurophysiology. , (2011).</li><li>Husch, A., Cramer, N., Harris-Warrick, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long+duration+perforated+patch+recordings+from+spinal+interneurons+of+adult+mice.">Long duration perforated patch recordings from spinal interneurons of adult mice.</a> Journal of Neurophysiology. , (2011).</li><li>Manuel, M., Zytnicki, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alpha,+beta+and+gamma+motoneurons:+functional+diversity+in+the+motor+system's+final+pathway.">Alpha, beta and gamma motoneurons: functional diversity in the motor system's final pathway.</a> J. Integr. Neurosci. 10, 243-276 (2011).</li><li>Nakanishi, S. T., Whelan, P. 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+decerebrate+adult+mouse+model+for+examining+the+sensorimotor+control+of+locomotion.">A decerebrate adult mouse model for examining the sensorimotor control of locomotion.</a> Journal of Neurophysiology. 107, 500-515 (2012).</li><li>Meehan, C. F., Grondahl, L., Nielsen, J. B., Hultborn, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fictive+locomotion+in+the+adult+decerebrate+and+spinal+mouse+in+vivo.">Fictive locomotion in the adult decerebrate and spinal mouse in vivo.</a> The Journal of Physiology. 590, 289-300 (2012).</li></ol>

Fremstillingen beskrevet her er den første, der tillader, i den voksne mus, samtidig intracellulær optagelse af en lumbal motoneuron og måling af den kraft, der produceres af muskelfibrene innerveret af dens axon. På grund af den lille størrelse af dyret, kan de kirurgiske færdigheder, der kræves for denne forberedelse være udfordrende at erhverve. Men når disse færdigheder beherskes, kan hele operationen udføres i tre timer, og dyrene kan overleve i op til 7 flere timer efter afs...

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> Navn på reagenset </td><td> Firma </td><td> Katalognummer </td><td> Kommentarer (valgfrit) </td></tr><tr><td> Atropinsulfat </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> Natriumpentobarbiton </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> Glucose </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> Blunt saks </td><td> FST </td><td> 14.079-10 </td><td></td></tr><tr><td> Blunt fine saks </td><td> FST </td><td> 15.025-10 </td><td></td></tr><tr><td> Vannas Spring Sakse </td><td> FST </td><td> 15.002-08 </td><td></td></tr><tr><td> Fin pincet savtakket </td><td> FST </td><td> 11.370-32 </td><td></td></tr><tr><td> Fin pincet savtakket </td><td> FST </td><td> 11.370-31 </td><td></td></tr><tr><td> Cunningham Spinal Adaptor </td><td> Stoelting Co </td><td></td><td></td></tr><tr><td> Kwik-Cast tætningsmiddel </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> Capnograph </td><td> CWE Inc </td><td> μcapstar </td><td></td></tr><tr><td> Varmetæppe </td><td> Harvard Apparatus </td><td> 507221F </td><td></td></tr><tr><td> Intracellulær forstærker </td><td> Axon Instruments </td><td> Axoclamp 2B </td><td></td></tr><tr><td> Pipette aftrækker </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 længe været et godt modelsystem til undersøgelse af neural funktion fordi det er en neuron i centralnervesystemet med de enestående egenskaber af (1) med let identificerbare mål (muskelfibrene), og derfor har en meget kendt funktion (at styre muskelkontraktion), (2) er den konvergerende mål for mange spinal-og nedadgående net, deraf navnet &quot;endelig fælles vej&quot;, og (3) have et stort soma, som gør det muligt at penetrere dem med skarpe intracellulære elektroder . Når endvidere undersøgt in vivo, er det muligt at optage samtidig den elektriske aktivitet af motoneuroner og den kraft udviklet af deres muskel mål. Udførelse af intracellulære optagelser af motoneuroner in vivo derfor sætte experimentalist i den unikke situation at kunne studere på samme tid, alle rummene i &quot;motorenhed&quot; (navnet givet til motoneuron, dens axon, ogmuskelfibrene den innerverer 1): input støder imod det motoneuron, de elektrofysiologiske egenskaber motoneuron, og virkningen af disse egenskaber på den fysiologiske funktion af de motoneuroner, dvs den kraft der produceres af dets motorenhed. Men denne fremgangsmåde er meget vanskelig på grund præparatet ikke kan blive lammet og dermed den mekaniske stabilitet for den intracellulære optagelse reduceres. Således har denne type forsøg kun opnået hos katte og rotter. Imidlertid kunne studiet af spinale motoriske systemer gør en formidabel spring, hvis det var muligt at udføre lignende forsøg i normale og genetisk modificerede mus. Af tekniske grunde har studiet af cellerne i rygmarven hos mus meste været begrænset til neonatal in vitro præparater, hvor motoneuroner og de ​​spinale netværk er umodne, er motoneuroner adskilt fra deres mål, og når undersøgt i skiver, den motoneurons er adskilt fra de fleste af deres input. Indtil for nylig havde kun et par grupper formået at udføre intracellulære optagelser af motoneuroner in vivo 2-4, herunder vores team, der offentliggjorde en ny præparat som tilladt os at opnå meget stabile optagelser af motoneuroner in vivo i voksne mus 5,6. Imidlertid blev disse optagelser opnået i lammede dyr, altså uden mulighed for at registrere den kraft Outputtet fra disse motoneuroner. Her præsenterer vi en forlængelse af denne oprindelige præparat, hvor vi var i stand til at opnå samtidige optagelser af de elektrofysiologiske egenskaber motoneuroner og af den kraft, der er udviklet af deres motorenhed. Dette er et vigtigt resultat, da det giver os mulighed for at identificere de forskellige typer af motoneuroner baseret på deres force profil, og derved afsløre deres funktion. Kombineret med genetiske modeller forstyrrer spinal segmental kredsløb 7-9, eller reproducting human disease 10,11, vi forventer, at denne teknik for at være et vigtigt redskab for studiet af spinal motoriske system.

Figur 1 viser, hvordan man identificere en motoneuron fra Triceps surae gruppen efter penetration. Ved lav stimulation intensitet, kan kun en monosynaptiske EPSP observeres (figur 1A). Ved højere intensitet, kan EPSP være stor nok til at udløse en &quot;orthodromic&quot; spids (figur 1B). Ved endnu højere stimulation intensitet, vises et alt-eller-intet antidromic pig, med en kortere forsinkelse end den monosynaptiske EPSP (figur 1C). Hvis tilstræk...

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 fremgangsmåde muliggør den samtidige intracellulære optagelse af en enkelt voksen mus motoneuron og måling af den kraft produceret af dets muskelfibre. Den kombinerede undersøgelse af de elektriske og mekaniske egenskaber af motoriske enheder i normale og genetisk modificerede dyr er et gennembrud for studiet af det neuromuskulære system.

Samtidig Intracellulær Optagelsen af ​​en Lumbar Motoneuron og styrkens Produceret af sin Motor Unit i den voksne mus<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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