Name: optic nerve transection: a model of adult neuron apoptosis in the central nervous system
Uploaded: 2011-05-12T12:00:00
Description: Watch this Scientific Journal Video about Optic Nerve Transection: A Model of Adult Neuron Apoptosis in the Central Nervous System at JoVE.com

PDK stöds av en CIHR driftsbidrag (MOP 86.523)

1. Kirurgisk teknik <ol><li> Experiment bör utföras med aseptisk teknik och efter protokollen djuret använda dina specifika institutionen. Instrument och material (lösningar, testa ämnen, spårämnen, nålar, etc.) kommer i kontakt med levande vävnad måste vara steril för att förhindra infektion och negativa effekter på djurens välbefinnande och potentiella negativa effekter på studien. </li></ol> 2. Anestesi <ol><li> Råttor blir nedsövd med hjälp av en veterinär isofluran Vap...

     

<ol>
	<li>Bahr, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live+or+let+die+-+retinal+ganglion+cell+death+and+survival+during+development+and+in+the+lesioned+adult+CNS.">Live or let die - retinal ganglion cell death and survival during development and in the lesioned adult CNS.</a> Trends Neurosci. 23, 483-4890 (2000).</li><li>Isenmann, S., Kretz, A., Cellerino, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+determinants+of+retinal+ganglion+cell+development,+survival,+and+regeneration.">Molecular determinants of retinal ganglion cell development, survival, and regeneration.</a> Prog Retin Eye Res. 22, 483-543 (2003).</li><li>Koeberle, P. D., Bahr, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+and+guidance+cues+for+regenerating+axons:+where+have+they+gone.">Growth and guidance cues for regenerating axons: where have they gone.</a> J Neurobiol. 59, 162-180 (2004).</li><li>Weishaupt, J. H., Bahr, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Degeneration+of+axotomized+retinal+ganglion+cells+as+a+model+for+neuronal+apoptosis+in+the+central+nervous+system+-+molecular+death+and+survival+pathways.">Degeneration of axotomized retinal ganglion cells as a model for neuronal apoptosis in the central nervous system - molecular death and survival pathways.</a> Restor. Neurol. Neurosci. 19, 1-2 (2001).</li><li>Valenzuela, G. a. r. c. i. a., E, S. C. S. h. a. r. m. a. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rescue+of+retinal+ganglion+cells+from+axotomy-induced+apoptosis+through+TRK+oncogene+transfer.">Rescue of retinal ganglion cells from axotomy-induced apoptosis through TRK oncogene transfer.</a> Neuroreport. 9, 3165-3170 (1998).</li><li>Kugler, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transduction+of+axotomized+retinal+ganglion+cells+by+adenoviral+vector+administration+at+the+optic+nerve+stump:+an+in+vivo+model+system+for+the+inhibition+of+neuronal+apoptotic+cell+death.">Transduction of axotomized retinal ganglion cells by adenoviral vector administration at the optic nerve stump: an in vivo model system for the inhibition of neuronal apoptotic cell death.</a> Gene Ther. 6, 1759-1767 (1999).</li><li>Lingor, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Down-regulation+of+apoptosis+mediators+by+RNAi+inhibits+axotomy-induced+retinal+ganglion+cell+death+in+vivo.">Down-regulation of apoptosis mediators by RNAi inhibits axotomy-induced retinal ganglion cell death in vivo.</a> Brain. 128, 550-558 (2005).</li><li>Koeberle, P. D., Gauldie, J., Ball, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+adenoviral-mediated+gene+transfer+of+interleukin-10,+interleukin-4,+and+transforming+growth+factor-beta+on+the+survival+of+axotomized+retinal+ganglion+cells.">Effects of adenoviral-mediated gene transfer of interleukin-10, interleukin-4, and transforming growth factor-beta on the survival of axotomized retinal ganglion cells.</a> Neuroscience. 125, 903-920 (2004).</li><li>Berkelaar, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axotomy+results+in+delayed+death+and+apoptosis+of+retinal+ganglion+cells+in+adult+rats.">Axotomy results in delayed death and apoptosis of retinal ganglion cells in adult rats.</a> J Neurosci. 14, 4368-4374 (1994).</li><li>Villegas-Perez, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influences+of+peripheral+nerve+grafts+on+the+survival+and+regrowth+of+axotomized+retinal+ganglion+cells+in+adult+rats.">Influences of peripheral nerve grafts on the survival and regrowth of axotomized retinal ganglion cells in adult rats.</a> J Neurosci. 8, 265-280 (1988).</li><li>Villegas-Perez, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+and+protracted+phases+of+retinal+ganglion+cell+loss+follow+axotomy+in+the+optic+nerve+of+adult+rats.">Rapid and protracted phases of retinal ganglion cell loss follow axotomy in the optic nerve of adult rats.</a> J Neurobiol. 24, 23-36 (1993).</li><li>Quigley, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinal+ganglion+cell+death+in+experimental+glaucoma+and+after+axotomy+occurs+by+apoptosis.">Retinal ganglion cell death in experimental glaucoma and after axotomy occurs by apoptosis.</a> Invest Ophthalmol Vis Sci. 36, 774-786 (1995).</li><li>Thanos, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Specific+transcellular+carbocyanine-labelling+of+rat+retinal+microglia+during+injury-induced+neuronal+degeneration.">Specific transcellular carbocyanine-labelling of rat retinal microglia during injury-induced neuronal degeneration.</a> Neurosci Lett. 127, 108-1012 (1991).</li><li>Thanos, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Specific+transcellular+staining+of+microglia+in+the+adult+rat+after+traumatic+degeneration+of+carbocyanine-filled+retinal+ganglion+cells.">Specific transcellular staining of microglia in the adult rat after traumatic degeneration of carbocyanine-filled retinal ganglion cells.</a> Exp Eye Res. 55, 101-117 (1992).</li></ol>

Det finns många varianter av detta kirurgiska ingrepp och flera av de steg i detta protokoll inte är nödvändiga. Det är endast nödvändigt att dra tillbaka de muskler som overlie synnerven för att få tillgång till den nerven. Men detta resulterar i ett mycket begränsat arbetsutrymme runt nerven som gör det kritiska slutskedet av transection svårare. I vissa situationer är det önskvärt att transfektera cellerna från synnerven stubben och den ökade tillgången utrymme som ges genom dra tillbaka alla extra...

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optic nerve transection: a model of adult neuron apoptosis in the central nervous system

Retinal ganglion celler (RGC: er) är CNS-nervceller som utdata visuell information från näthinnan till hjärnan via synnerven. Synnerven kan nås inom bana i ögat och helt transected (axotomized), skära av axoner av hela RGC befolkningen. Synnerven transection är en reproducerbar modell av apoptotiska nervcellsdöd i vuxna CNS 1-4. Denna modell är särskilt attraktiva eftersom glaskroppen kammare i ögat fungerar som en kapsel för drug delivery till näthinnan, som möjliggör experimentella manipulationer via intraokulära injektioner. Spridning av kemikalier genom glaskroppen vätska ser till att de agerar på hela RGC befolkningen. Dessutom kan RGC: er vara selektivt transfekterade genom att korta interfererande RNA (siRNA), plasmider eller virala vektorer på den avskurna ändan av synnerven 5-7 eller injicera vektorer i deras mål, den överlägsna colliculi 8. Detta gör det möjligt för forskare att studera apoptotiska mekanismer i önskad neuronala befolkningen utan störande effekter på andra åskådare nervceller eller omgivande Glia. En ytterligare fördel är den lätthet och precision med vilken cellöverlevnad kan kvantifieras efter skada. Näthinnan är en platt, lager vävnad och RGC: er är lokaliserade i de innersta lager, ganglion cellager. Överlevnad RGC: er kan spåras över tid genom att tillämpa en fluorescerande spårämne (3% Fluorogold) på den avskurna ändan av synnerven vid axotomy, eller genom att injicera spårgas i den överlägsna colliculi (RGC mål) en vecka före axotomy. Den spårämne är retrogradely transporteras, märkning hela RGC befolkningen. Eftersom gangliecell lagret är ett monolager (en cell tjock), kan RGC densiteter kvantifieras i platta monterad vävnad, utan behov av stereology. Synnerven transection leder till apoptotisk död 90% av skadade RGC: er inom 14 dagar postaxotomy 9-11. RGC apoptos har en karakteristisk tid-kurs där celldöd är försenad 3-4 dagar postaxotomy, varefter cellerna snabbt urarta. Detta ger ett tidsfönster för experimentella manipulationer som riktas mot involverade i apoptos.

Watch this Scientific Journal Video about Optic Nerve Transection: A Model of Adult Neuron Apoptosis in the Central Nervous System at JoVE.com

Optic Nerve Transection: A Model of Adult Neuron Apoptosis in the Central Nervous System

Synnerven transection är en allmänt använd modell av vuxna CNS-skada. Nittio procent av retinal ganglion celler (RGC: er), vars axoner är helt transected (axotomy) dör inom 14 dagar efter axotomy. Denna modell är lätta att experimentella manipulationer och mycket reproducerbar.

Synnerven transection: En modell av Vuxen Neuron apoptos i det centrala nervsystemet

Retinal ganglion cells (RGCs) are CNS neurons that output visual information from the retina to the brain, via the optic nerve. 
The optic nerve can be ...

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Traditionally, tissue visualization has required that the tissue of interest be serially sectioned and imaged, subjecting each tissue section to unique ...