Name: tibial nerve transection - a standardized model for denervation-induced skeletal muscle atrophy in mice
Uploaded: 2013-11-03T19:00:00
Description: Watch this Scientific Journal Video about Tibial Nerve Transection - A Standardized Model for Denervation-induced Skeletal Muscle Atrophy in Mice at JoVE.com

Dette arbejde blev støttet af tilskud fra CIHR Neuromuskulær Research Partnership (JNM - 90.959, til JAEB).

Forud for at bruge denne model, skal efterforskerne har fået godkendt den kirurgiske protokol fra deres institutions brug af dyr styrende organ. Modellen er godkendt af Research Ethics Board, Hamilton Health Sciences Corporation, McMaster University (AUP # 10-04-24) og udføres i nøje overensstemmelse med anbefalingerne fra den canadiske Råd om Animal Care. 1.. Mus Forberedelse <ol><li> Afvejes musen. Fremkald anæstesi med 5% isofluran eller 2% halothan. Den anvendte kredsløb bør sik...

<ol>
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Den tibiale nerve transection model af denervering-induceret skeletmuskel atrofi er en almindeligt anvendt og godt valideret model i rotter. Vi har tilpasset denne model til brug i mus, som giver investigator til at drage fordel af eksistensen af gensplejsede mus og studere processen muskelatrofi in vivo i fravær af proteiner er afgørende for reguleringen af muskelmasse 7,8. Gastrocnemius og soleus muskler, både denerveres i denne model, kan være let og hurtigt dissekeret med minimal håndtering,...

Skeletmuskulatur denervering, grundet traumatisk perifer nerveskade, sygdom eller farmakologiske indgreb, resulterer i øjeblikkelig tab af muskel frivillig kontraktile funktion. Muscle samtidig begynder at atrofi og denne atrofi er reversibel, hvis rettidig, god kvalitet reinnervation opstår 1,2. I mangel af reinnervation, skrider myofiber atrofi og irreversible biologiske forandringer i musklen forekomme med muskel fibrose og myofiber død. Her har vi demonstrere tibial nerve transection model, en model af denervering-induceret skeletmuskulatur atrofi og fibrose, i mus. Denne model gør det muligt for forskerne at undersøge de fysiologiske, cellulære og molekylære biologiske mekanismer, der ligger til grund muskelatrofi in vivo i gastrocnemius og soleus muskler. Mens historisk brugt overvejende rotter nyere anvendelse af denne model på knockout og transgene mus linjer specifikt giver efterforskerne at vurdere den rolle deresbestemt protein (erne) af interesse i induktionen, udvikling og vedligeholdelse, eller alternativt opløsning, muskelatrofi og fibrose in vivo. Den tibiale nerve er en blandet motor-sensorisk perifer nerve hos gnavere bagben, og er en af ​​de tre terminale grene af iskiasnerven. Overskæring af den tibiale nerve denervates gastrocnemius, soleus og plantaris muskler (og de ​​tre små dybe bøjemuskler af foden, herunder tibialis posterior, flexor digitorum longus og flexor hallicus longus), og er et godt standardiseret og valideret model i rotter 3,4 . Gastrocnemius og soleus muskler kan nemt dissekeret serielle tidspunkter efter tibial nerve transection, fikseret og forarbejdet til vurdering af muskel histologi og muskelfibrene morphometrics eller lynfrosset til ekstraktion af muskel-RNA og protein med henblik på at studere, for eksempel, de cellulære signalnetværker regulerer muskelatrofi. Den gastrocnemius muskel er en blandet fibertype muskel (type I og type II, selvom overvejende type II) og soleusmusklen er sammensat af en stor del af type I-fibre, hvorved der tilvejebringes både hurtig og langsom twitch muskel til vurdering 5,6. Den tibial nerve transection Modellen er velegnet til at studere processen denervering-induceret muskelatrofi på både kort sigt (dage) 7 og lang sigt (uger til måneder) 4,8. I modsætning til iskiasnerven overskæring model (en anden model af denervering-induceret muskelatrofi almindeligt anvendt i gnavere), inducerer tibial nerve transection mindre morbiditet i dyret, hvilket gør det mere tiltrækkende model. Transection af iskiasnerven denervates alle musklerne i benet (under knæet) og fod, der krænker dyrets evne til at ambulate 2, mens transection af tibial nerve forlader peroneal og sural nerve grene af iskiasnerven intakt, således at beskyttederes mål muskler og sensoriske territorier. Musen er i stand til plantar flex eller vend foden, men er i stand til at ambulate nemt og vægten bærer ligeligt på begge bagben, og dermed betydeligt mindske sygeligheden af ​​modellen. Ganganalyse studier, som evaluerede walking mønstre er blevet udført i rotter efter tibiale og iskiasnerven skader og vise, at fodaftryk og vægtbærende bedre bevaret med tibial skade 9,10. Desuden, i tibial nerve transection model kan peronealnerve mobiliseres på et senere tidspunkt og overføres som en kilde til forsinket reinnervation, hvis undersøgelsens design kræver 3. I modsætning hertil nødvendiggør forsinket reinnervation i iskiasnerven transection model brugen af ​​en nerve graft til iskiasnerven underskud, meget væsentligt at øge tekniske vanskeligheder af modellen og begrænse brugen til dygtige kirurger. Mens den tibiale nerve transection model requires kendskab for operatøren med steril operative teknik i dyre kirurgi, både tibial nerve og lægmuskler den innerverer er let tilgængelige og identificerbare for manipulation, så personer, som ikke kirurger, eller meget erfarne med dyr operation, let kan mestre denne model .

<table border="1">
	<tbody>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Reagents and Materials</td>
		</tr>
		<tr>
			<td>10-0 Nylon suture</td>
			<td>Ethicon</td>
			<td>2850G</td>
			<td></td>
		</tr>
		<tr>
			<td>5-0 Vicryl suture</td>
			<td>Ethicon</td>
			<td>J553G</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Equipment</td>
		</tr>
		<tr>
			<td>Spring microdissecting scissors</td>
			<td>Fine Surgical Tools</td>
			<td>15021-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultra fine forceps</td>
			<td>Fine Surgical Tools</td>
			<td>11370-40</td>
			<td></td>
		</tr>
		<tr>
			<td>Non locking micro needle holder (driver)</td>
			<td>Fine Surgical Tools</td>
			<td>12076-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Spring retractor</td>
			<td>Fine Surgical Tools</td>
			<td>17000-02</td>
			<td></td>
		</tr>
	</tbody>
</table>

tibial nerve transection - a standardized model for denervation-induced skeletal muscle atrophy in mice

Den tibiale nerve transection model er en veltolereret, valideret og reproducerbar model af denervering-induceret skeletmuskulatur atrofi i gnavere. Selv om de oprindeligt udviklet og anvendt i udstrakt grad i rotter på grund af sin større størrelse, den tibiale nerve i mus er stort nok, at det let kan manipuleres med enten knuse eller overskæring, forlader peroneal og sural nerve grene af iskiasnerven intakt og dermed bevare deres mål muskler. Således er denne model giver fordelene ved at inducere mindre sygelighed og hindring af ambulation end iskiasnerven overskæring model og også tillader undersøgere at studere de fysiologiske, cellulære og molekylære biologiske mekanismer, der regulerer processen muskelatrofi i gensplejsede mus. Den tibial nerve leverer gastrocnemius, soleus og plantaris muskler, så dens transection tillader studiet af denerverede skeletmuskulatur bestående af hurtige ryk type II fibre og / eller langsomme twitch type Ifibre. Her har vi demonstrere tibial nerve transection model i C57Black6 mus. Vi vurderer atrofi af musculus gastrocnemius, som en repræsentativ muskel, ved 1, 2 og 4 uger efter denervering ved måling muskel vægt og fibertype specifik tværsnitsareal på paraffinindstøbte histologiske snit immunfarvet for hurtige ryk myosin.

<html> Tibial nerve transection denervates gastrocnemius, soleus og plantaris muskler i læggen. Her vi vurdere udviklingen af ​​atrofi i musculus gastrocnemius, som en repræsentativ muskel. Musculus gastrocnemius blev høstet fra 2-3 måneder gammel C57Black 6 mus (Jackson Laboratories) denerverede for 1, 2 eller 4 uger. Muskel vægte gradvist falde (figur 1), som gør tværsnitsarealet af type II hurtige ryk muskelfibre (figur 2), over tid. Gastrocnemius er en blandet fibertype mu...

Watch this Scientific Journal Video about Tibial Nerve Transection - A Standardized Model for Denervation-induced Skeletal Muscle Atrophy in Mice at JoVE.com

Tibial Nerve Transection - A Standardized Model for Denervation-induced Skeletal Muscle Atrophy in Mice

Den tibiale nerve transection model er en veltolereret, valideret og reproducerbar model af skeletmuskulaturen atrofi. Modellen kirurgiske protokol er beskrevet og vist i C57Black6 mus.

Tibial Nerve transection - En standardiseret model for denervering-induceret Skeletmuskel Atrofi hos mus

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. ...

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