Name: repeated measurement of respiratory muscle activity and ventilation in mouse models of neuromuscular disease
Uploaded: 2017-04-17T14:00:00
Description: Watch this Scientific Journal Video about Repeated Measurement of Respiratory Muscle Activity and Ventilation in Mouse Models of Neuromuscular Disease at JoVE.com

Støtte til dette arbejde blev leveret af en Cincinnati Children &#39;s Hospital Medical Center administrator Award til SAC og en NIH træning tilskud (T32NS007453) til VNJ

Eksperimentelle procedurer blev godkendt af Cincinnati Children &#39;s Hospital Medical Center Institutional Animal Care og brug Udvalg og gennemføres i overensstemmelse med NIH Guide til pleje og anvendelse af forsøgsdyr. 1. Forberedelse til telemetri Device implantat kirurgi <ol><li> Sæt på personlige værnemidler (dvs. scrubs, skoovertræk, kjole, hår netto, maske, og kirurgiske handsker). BEMÆRK: Denne operation kræver et sterilt område. </li><li> Tænd inkubator (servostyret luftfu...

<ol>
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H., 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=Accessory+respiratory+muscles+enhance+ventilation+in+ALS+model+mice+and+are+activated+by+excitatory+V2a+neurons.">Accessory respiratory muscles enhance ventilation in ALS model mice and are activated by excitatory V2a neurons.</a> Exp Neurol. 287 (Pt. 2, 192-204 (2017).</li><li>Moldovan, 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=Nerve+excitability+changes+related+to+axonal+degeneration+in+amyotrophic+lateral+sclerosis:+Insights+from+the+transgenic+SOD1(G127X)+mouse+model.">Nerve excitability changes related to axonal degeneration in amyotrophic lateral sclerosis: Insights from the transgenic SOD1(G127X) mouse model.</a> Exp Neurol. 233 (1), 408-420 (2012).</li><li>Pagliardini, S., Gosgnach, S., Dickson, C. 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Proceduren demonstreret her tillader noninvasiv (efter indledende kirurgisk implantation af senderen) måling af respiratorisk muskelaktivitet og ventilation gennem mange måneder i det samme dyr. Denne teknik har adskillige fordele i forhold til standard EMG teknikker i bedøvede mus: 1) forsøgene kræver færre mus og tilvejebringer evnen til at optage data fra det samme sted i en enkelt mus tværs sygdomsstadier (i stedet for at bruge flere mus på forskellige sygdomstilstande stadier); 2) data analyse kan udføres ...

Tilbehør respiratoriske muskler (ARM&#39;er) øge ventilationen i perioder med høj efterspørgsel (fx motion) og hjælpe med at bevare ventilation, når membranen funktion er kompromitteret efter skade eller sygdom 1, 2. Selvom ændringer i membran funktion er blevet godt beskrevet i amyotrofisk lateral sklerose (ALS) patienter og musemodeller 3, 4, 5, 6, meget mindre om aktiviteten eller funktionen af rentetilpasningslånene i ALS. , En undersøgelse foreslog imidlertid, at ALS-patienter, der rekrutterer våben har en bedre prognose end dem med lignende membran dysfunktion, der ikke gør 7. Endvidere ARM aktivitet er tilstrækkelig til respiration i tilfælde af mellemgulvet lammelse 8. Disse undersøgelser viser, at strategier at forøge ARM-funktionen kan forbedre breathing i patienter, der lider neuromuskulær sygdom, rygmarvsskade, eller andre tilstande, hvor membranen funktion er nedsat. Men de mekanismer, der styrer ARM rekruttering til at trække vejret er stort set ukendt. Metoder til måling af åndedrætsfunktionen og ændringer i ARM aktivitet over tid i dyremodeller af sygdom eller skade er nødvendige for at undersøge, hvordan rentetilpasningslån rekrutteres, samt at evaluere behandlingsformer for at forbedre ARM rekruttering og ventilation. Endvidere kan øget aktivitet af våben falder sammen med det progressive tab af membranen funktion være et nyttigt biomarkør for sygdomsprogression i neuromuskulære sygdomme, såsom ALS 7, 9, 10. Denne protokol beskriver en fremgangsmåde til ikke-invasiv (efter den første kirurgi) og gentagne gange måle aktiviteten af ​​respiratoriske muskler og ventilation i vågne, opfører mus. Synkroniserede optagelser af elektromyografy (EMG), hel-legemsplethysmografi (WBP), og video tillader undersøgeren for at vurdere, hvordan ændringer i ARM aktivitet indvirkning ventilation og til at bestemme, når motivet er i hvile eller bevæger sig. En stor fordel ved denne fremgangsmåde er, at den kan udføres i vågne, opfører mus, mens nogle alternative metoder til måling af EMG kræver anæstesi og / eller er terminalprocedurer 11, 12, 13. Registrering af EMG-aktivitet i vågen mus over tid kan også opnås gennem den kroniske implantation af EMG fører, hvor musen er tøjret med ledninger til erhvervelse systemet 14, 15. Fordi tøjring en mus kan forstyrre normal bevægelse eller adfærd og kan ikke være foreneligt med en standard plethysmografi kammer, den beskrevne metode benyttes telemetri enheder til trådløst at overføre EMG-signalet til erhvervelse system. Senderen kantændes eller slukkes med en magnet for at spare på batteriet og giver gentagne målinger af EMG aktivitet over flere måneder. Denne protokol kan let tilpasses til at måle aktiviteten af ​​yderligere respiratoriske eller ikke-respiratoriske muskler ved at indsætte EMG fører ind forskellige muskler. Alternativt kan en af de to ledninger anvendes til at måle EEG aktivitet at vurdere søvntilstand eller til at identificere anfaldsaktivitet 16. Denne teknik har med succes været anvendt til at måle ændringer i ARM aktivitet i hvile i hele sygdomsprogression i en musemodel af ALS og at identificere de vigtigste neuroner kørsel ARM aktivitet i raske mus 10.

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			<td>B6.Cg-Tg (SOD1*G93A)1 Gur/J</td>
			<td>Jackson Laboratory</td>
			<td>4435</td>
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			<td>Buxco Respiratory Products/ Data Sciences International</td>
			<td>601-1425-001</td>
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			<td>Telemetry Receivers (Model RPC-1)</td>
			<td>Data Sciences International</td>
			<td>272-6001-001</td>
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			<td>Bias Flow Pump (Model BFL0500)</td>
			<td>Data Sciences International</td>
			<td>601-2201-001</td>
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			<td>ACQ-7000 USB</td>
			<td>Data Sciences International</td>
			<td>PNM-P3P-7002XS</td>
			<td></td>
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			<td>Dataquest A.R.T. Data Exchange Matrix</td>
			<td>Data Sciences International</td>
			<td>271-0117-001</td>
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			<td>New Ponemah Analysis System</td>
			<td>Data Sciences International</td>
			<td>PNM-POST-CFG</td>
			<td></td>
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			<td>Ponemah Physiology Platform Acqusition software v5.20</td>
			<td>Data Sciences International</td>
			<td>PNM-P3P-520</td>
			<td></td>
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			<td>Ponemah Unrestrained Whole Breath Plethysmography analysis package v5.20</td>
			<td>Data Sciences International</td>
			<td>PNM-URP100W</td>
			<td></td>
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			<td>Configured Ponemah Software System</td>
			<td>Data Sciences International</td>
			<td>PNM-P3P-CFG</td>
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			<td>Analysis Module (URP)</td>
			<td>Data Sciences International</td>
			<td>PNM-URP100W</td>
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			<td>Universal Amplifier</td>
			<td>Data Sciences International</td>
			<td>13-7715-59</td>
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			<td>Sync Board</td>
			<td>Data Sciences International</td>
			<td>271-0401-001</td>
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			<td>Sync Cable</td>
			<td>Data Sciences International</td>
			<td>274-0030-001</td>
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			<td>Transducer-Pressure Buxco</td>
			<td>Data Sciences International</td>
			<td>600-1114-001</td>
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			<td>Flow Meter</td>
			<td>Data Sciences International</td>
			<td>600-1260-001</td>
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			<td>Magnet and Radio included in F20-EET Starter Kit</td>
			<td>Data Sciences International</td>
			<td>276-0400-001</td>
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			<td>Axis P1363 Video Camera&#160;&#160;</td>
			<td>Data Sciences International</td>
			<td>275-0201-001</td>
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			<td>Fisher Scientific</td>
			<td>50-821-785</td>
			<td>Enzyme Detergent</td>
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			<td>Minntech Corporation</td>
			<td>78337-000</td>
			<td>Chemical Sterilant</td>
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			<td>Stereo Dissecting Microscope (Model MEB126)</td>
			<td>Leica</td>
			<td>10-450-508</td>
			<td></td>
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			<td>Servo-Controlled Humidifier/Infant Incubator</td>
			<td>OHMEDA Ohio Care Plus</td>
			<td>6600-0506-803</td>
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			<td>TL11M2-F20-EET Transmitters</td>
			<td>Data Sciences International</td>
			<td>270-0124-001</td>
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			<td>Dumont #2 Laminectomy Forceps - Standard Tips/Straight/12cm (x2)&#160;</td>
			<td>Fine Scientific Instruments</td>
			<td>11223-20</td>
			<td>For handling wires</td>
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			<td>Fine Scientific Instruments</td>
			<td>11223-20</td>
			<td>For surgery</td>
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			<td>Narrow Pattern Forceps- Serrated/Curved/12cm</td>
			<td>Fine Scientific Instruments</td>
			<td>17003-12</td>
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			<td>Spring Scissors - Tough Cut/Straight/Sharp/12.5cm/6mm Cutting Edge</td>
			<td>Fine Scientific Instruments</td>
			<td>15124-12</td>
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			<td>Fine Scientific Instruments</td>
			<td>14072-10</td>
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			<td>Fine Scientific Instruments</td>
			<td>14058-11</td>
			<td>For cutting wires and clipping nails</td>
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			<td>World Precision Instruments</td>
			<td>500236</td>
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			<td>Fine Scientific Instruments</td>
			<td>10010-00</td>
			<td>For preparing lead caps</td>
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			<td>Coviden</td>
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			<td>Zoetis Inc.</td>
			<td>6606-65-1</td>
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			<td>Vitality Medical</td>
			<td>118030055</td>
			<td></td>
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			<td>Becton Dickinson and Co.</td>
			<td>305-145</td>
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			<td>Henry Schein Animal Health</td>
			<td>100-9175</td>
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			<td>Andis</td>
			<td>049-06-0271</td>
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			<td>Henry Schein Animal Health</td>
			<td>29404</td>
			<td>Anesthetic&#160;</td>
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			<td>Priority Care 1</td>
			<td>MS070PC</td>
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			<td>Butler Schein</td>
			<td>55482</td>
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			<td>Henry Schein Animal Health</td>
			<td>48272</td>
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			<td>Dow Corning Corporation</td>
			<td>1597418</td>
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			<td>JorVet</td>
			<td>JOR784BN</td>
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repeated measurement of respiratory muscle activity and ventilation in mouse models of neuromuscular disease

Tilbehør respirationsmusklerne bidrage til at bevare ventilation, når membranen er svækket. Den følgende protokol beskriver en fremgangsmåde til gentagne målinger over uger eller måneder efter tilbehør respiratorisk muskelaktivitet samtidig måler ventilation i en ikke-bedøvet, frit opfører mus. Teknikken omfatter kirurgisk implantation af en radiosender og indsættelse af elektroden fører ind i scalene og trapezius muskler til at måle elektromyogram aktivitet af disse inspiratoriske muskler. Ventilation måles ved hel-legemsplethysmografi, og dyrs bevægelser vurderes ved video og er synkroniseret med electromyogram aktivitet. Målinger af muskelaktivitet og ventilation i en musemodel for amyotrofisk lateral sklerose præsenteres for at vise, hvordan dette værktøj kan anvendes til at undersøge, hvordan respiratorisk muskelaktivitet ændringer over tid og at vurdere virkningen af ​​muskelaktivitet på ventilation. De beskrevne metoder kan easily tilpasses til at måle aktiviteten af ​​andre muskler eller vurdere tilbehør respiratorisk muskel aktivitet i yderligere musemodeller af sygdom eller tilskadekomst.

Den beskrevne protokol blev anvendt til at implantere en telemetrienhed og registrere scalenii og trapezius EMG, WBP, og video af en SOD1 (G93A) ALS model mus. Perioder, hvor dyret er inaktivt (for eksempel bevæger sig ikke) blev identificeret ved anvendelse videooptagelsen og bekræftet ved den manglende bevægelse-relaterede aktiviteter i WBP trace (figur 3A). Inaktive indbefatter tid tilbragt i REM eller ikke-REM søvn, såvel som tid brugt vågen men stadig...

Watch this Scientific Journal Video about Repeated Measurement of Respiratory Muscle Activity and Ventilation in Mouse Models of Neuromuscular Disease at JoVE.com

Repeated Measurement of Respiratory Muscle Activity and Ventilation in Mouse Models of Neuromuscular Disease

Papiret introducerer en fremgangsmåde til gentagne målinger af ventilation og respiratorisk muskelaktivitet i en frit opfører amyotrofisk lateral sklerose (ALS) musemodel hele sygdomsprogression med hel-legemsplethysmografi og elektromyografi via en implanteret telemetrienheden.

Gentagen Måling af åndedrætsorganer muskel aktivitet og ventilation i musemodeller af Neuromuskulær sygdom

Accessory respiratory muscles help to maintain ventilation when diaphragm function is impaired. The following protocol describes a method for repeated ...

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