Name: in vivo imaging of optic nerve fiber integrity by contrast-enhanced mri in mice
Uploaded: 2014-07-22T14:00:00
Description: Watch this Scientific Journal Video about In vivo Imaging of Optic Nerve Fiber Integrity by Contrast-Enhanced MRI in Mice at JoVE.com

AK er støttet af Oppenheim Foundation og RH er støttet af Velux Fonden. Vi takker I. Krumbein til teknisk og K. Buder for histologisk support og J. Goldschmidt (Leibniz Institute for neurobiologi, Magdeburg, Tyskland) for teknisk rådgivning om TIMM farvning.

Alle animalske indgreb udføres i overensstemmelse med den europæiske konvention til Animal Care og brugen af forsøgsdyr og ARVO erklæring til brug af dyr i Ophthalmic og Vision Research. Alle forsøg er godkendt af den lokale etiske komité. Proceduren for ON skade i mus er beskrevet andetsteds 9. 1.. Intravitreale Mangan Injection <ol><li> Udfør Mn 2 + indsprøjtning 24 timer forud for MR-scanning ved hjælp af en assistent. Bedøve dyrene ved ...

<ol>
	<li>Kretz, A., 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=Simvastatin+promotes+heat+shock+protein+27+expression+and+Akt+activation+in+the+rat+retina+and+protects+axotomized+retinal+ganglion+cells+in+vivo.">Simvastatin promotes heat shock protein 27 expression and Akt activation in the rat retina and protects axotomized retinal ganglion cells in vivo.</a> Neurobiol Dis. 21, 421-430 (2006).</li><li>Lima, 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=Combinatorial+therapy+stimulates+long-distance+regeneration,+target+reinnervation,+and+partial+recovery+of+vision+after+optic+nerve+injury+in+mice.">Combinatorial therapy stimulates long-distance regeneration, target reinnervation, and partial recovery of vision after optic nerve injury in mice.</a> Int Rev Neurobiol. 106, 153-172 (2012).</li><li>Lima, 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=Full-length+axon+regeneration+in+the+adult+mouse+optic+nerve+and+partial+recovery+of+simple+visual+behaviors.">Full-length axon regeneration in the adult mouse optic nerve and partial recovery of simple visual behaviors.</a> Proc Natl Acad Sci U S A. 109, 9149-9154 (2012).</li><li>Goetze, B., 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=Vision+and+visual+cortical+maps+in+mice+with+a+photoreceptor+synaptopathy:+reduced+but+robust+visual+capabilities+in+the+absence+of+synaptic+ribbons.">Vision and visual cortical maps in mice with a photoreceptor synaptopathy: reduced but robust visual capabilities in the absence of synaptic ribbons.</a> Neuroimage. 49, 1622-1631 (2010).</li><li>Luo, X., 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=Three-dimensional+evaluation+of+retinal+ganglion+cell+axon+regeneration+and+pathfinding+in+whole+mouse+tissue+after+injury.">Three-dimensional evaluation of retinal ganglion cell axon regeneration and pathfinding in whole mouse tissue after injury.</a> Exp Neurol. 247, 653-662 (2013).</li><li>Pautler, R. G., 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=In+vivo+neuronal+tract+tracing+using+manganese-enhanced+magnetic+resonance+imaging.">In vivo neuronal tract tracing using manganese-enhanced magnetic resonance imaging.</a> Magn Reson Med. 40, 740-748 (1998).</li><li>Watanabe, T., 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=Mapping+of+retinal+projections+in+the+living+rat+using+high-resolution+3D+gradient-echo+MRI+with+Mn2+-induced+contrast.">Mapping of retinal projections in the living rat using high-resolution 3D gradient-echo MRI with Mn2+-induced contrast.</a> Magn Reson Med. 46, 424-429 (2001).</li><li>Pautler, R. G. <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,+trans-synaptic+tract-tracing+utilizing+manganese-enhanced+magnetic+resonance+imaging+(MEMRI).">In vivo, trans-synaptic tract-tracing utilizing manganese-enhanced magnetic resonance imaging (MEMRI).</a> NMR biomed. 17, 595-601 (2004).</li><li>Haenold, R., 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=Magnetic+resonance+imaging+of+the+mouse+visual+pathway+for+in+vivo+studies+of+degeneration+and+regeneration+in+the+CNS.">Magnetic resonance imaging of the mouse visual pathway for in vivo studies of degeneration and regeneration in the CNS.</a> Neuroimage. 59, 363-376 (2012).</li><li>Lindsey, J. D., 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=Magnetic+resonance+imaging+of+the+visual+system+in+vivo:+transsynaptic+illumination+of+V1+and+V2+visual+cortex.">Magnetic resonance imaging of the visual system in vivo: transsynaptic illumination of V1 and V2 visual cortex.</a> Neuroimage. 34, 1619-1626 (2007).</li><li>Bearer, E. L., 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=Role+of+neuronal+activity+and+kinesin+on+tract+tracing+by+manganese-enhanced+MRI+(MEMRI).">Role of neuronal activity and kinesin on tract tracing by manganese-enhanced MRI (MEMRI).</a> Neuroimage. 37, S37-S46 (2007).</li><li>Mendonca-Dias, M. 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=Paramagnetic+contrast+agents+in+nuclear+magnetic+resonance+medical+imaging.">Paramagnetic contrast agents in nuclear magnetic resonance medical imaging.</a> Semin Nucl Med. 13, 364-376 (1983).</li><li>Thuen, 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=Manganese-enhanced+MRI+of+the+optic+visual+pathway+and+optic+nerve+injury+in+adult+rats.">Manganese-enhanced MRI of the optic visual pathway and optic nerve injury in adult rats.</a> J Magn Reson Imaging. 22, 492-500 (2005).</li><li>Sandvig, I., 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=In+vivo+MRI+of+olfactory+ensheathing+cell+grafts+and+regenerating+axons+in+transplant+mediated+repair+of+the+adult+rat+optic+nerve.">In vivo MRI of olfactory ensheathing cell grafts and regenerating axons in transplant mediated repair of the adult rat optic nerve.</a> NMR biomed. 25, 620-631 (2012).</li><li>Chan, K. 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=In+vivo+retinotopic+mapping+of+superior+colliculus+using+manganese-enhanced+magnetic+resonance+imaging.">In vivo retinotopic mapping of superior colliculus using manganese-enhanced magnetic resonance imaging.</a> Neuroimage. 54, 389-395 (2011).</li><li>Chan, K. 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=In+vivo+chromium-enhanced+MRI+of+the+retina.">In vivo chromium-enhanced MRI of the retina.</a> Magn Reson Med. 68, 1202-1210 (2012).</li><li>Herrmann, K. 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=Possibilities+and+limitations+for+high+resolution+small+animal+MRI+on+a+clinical+whole-body+3T+scanner.">Possibilities and limitations for high resolution small animal MRI on a clinical whole-body 3T scanner.</a> Magma. 25, 233-244 (2012).</li><li>Villegas-Perez, M. P., 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=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>Takahashi, Y., 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=Development+of+spontaneous+optic+neuropathy+in+NF-&#954;&#914;50-deficient+mice:+requirement+for+NF-&#954;&#914;p50+in+ganglion+cell+survival.">Development of spontaneous optic neuropathy in NF-&#954;&#914;50-deficient mice: requirement for NF-&#954;&#914;p50 in ganglion cell survival.</a> Neuropathol Appl Neurobiol. 33, 692-705 (2007).</li><li>Herrmann, K. H. P., 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=MRI+compatible+small+animal+monitoring+and+triggering+system+for+whole+body+scanners.">MRI compatible small animal monitoring and triggering system for whole body scanners.</a> Z Med Phys. 24, 55-64 (2013).</li><li>Danscher, G., Zimmer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+Timm+sulphide+silver+method+for+light+and+electron+microscopic+localization+of+heavy+metals+in+biological+tissues.">An improved Timm sulphide silver method for light and electron microscopic localization of heavy metals in biological tissues.</a> Histochemistry. 55, 27-40 (1978).</li><li>Angenstein, F., 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=Manganese-enhanced+MRI+reveals+structural+and+functional+changes+in+the+cortex+of+Bassoon+mutant+mice.">Manganese-enhanced MRI reveals structural and functional changes in the cortex of Bassoon mutant mice.</a> Cereb cortex. 17, 28-36 (2007).</li><li>Thuen, 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=Manganese-enhanced+MRI+of+the+rat+visual+pathway:+acute+neural+toxicity,+contrast+enhancement,+axon+resolution,+axonal+transport,+and+clearance+of+Mn(2).">Manganese-enhanced MRI of the rat visual pathway: acute neural toxicity, contrast enhancement, axon resolution, axonal transport, and clearance of Mn(2).</a> J Magn Reson Imaging. 28, 855-865 (2008).</li><li>Lehmann, K., 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=Vision+and+visual+plasticity+in+ageing+mice.">Vision and visual plasticity in ageing mice.</a> Restor Neurol Neurosci. 30, 161-178 (2012).</li><li>Takeda, A., 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=Manganese+transport+in+the+neural+circuit+of+rat+CNS.">Manganese transport in the neural circuit of rat CNS.</a> Brain Res Bull. 45, 149-152 (1998).</li><li>Nairismagi, J., 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=Manganese-enhanced+magnetic+resonance+imaging+of+mossy+fiber+plasticity+in+vivo.">Manganese-enhanced magnetic resonance imaging of mossy fiber plasticity in vivo.</a> Neuroimage. 30, 130-135 (2006).</li><li>Smith, K. D., 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=In+vivo+axonal+transport+rates+decrease+in+a+mouse+model+of+Alzheimer's+disease.">In vivo axonal transport rates decrease in a mouse model of Alzheimer's disease.</a> Neuroimage. 35, 1401-1408 (2007).</li><li>Berkowitz, B. A., 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=Noninvasive+and+simultaneous+imaging+of+layer-specific+retinal+functional+adaptation+by+manganese-enhanced+MRI.">Noninvasive and simultaneous imaging of layer-specific retinal functional adaptation by manganese-enhanced MRI.</a> Invest Ophthalmol Vis Sci. 47, 2668-2674 (2006).</li><li>Schnapf, J. L. B. D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+photoreceptor+cells+respond+to+light.">How photoreceptor cells respond to light.</a> Sci. Am. 256 (8), (1987).</li><li>Yu, X., 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=In+vivo+auditory+brain+mapping+in+mice+with+Mn-enhanced+MRI.">In vivo auditory brain mapping in mice with Mn-enhanced MRI.</a> Nat Neurosci. 8, 961-968 (2005).</li><li>Sun, S. W., 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=Noninvasive+topical+loading+for+manganese-enhanced+MRI+of+the+mouse+visual+system.">Noninvasive topical loading for manganese-enhanced MRI of the mouse visual system.</a> Invest Ophthalmol Vis Sci. 52, 3914-3920 (2011).</li><li>Sun, S. W., 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=Impact+of+repeated+topical-loaded+manganese-enhanced+MRI+on+the+mouse+visual+system.">Impact of repeated topical-loaded manganese-enhanced MRI on the mouse visual system.</a> Invest Ophthalmol Vis Sci. 53, 4699-4709 (2012).</li></ol>

MEMRI af det visuelle system udvider konventionelle neurobiologiske teknikker til vurdering af funktionalitet under naive og patologiske tilstande. Bortset fra at levere et unikt indblik i integriteten af en isoleret CNS fiber-tarmkanalen, kan MEMRI let suppleres med adfærdsmæssige test, fx optometri og visuelt baserede vand opgaver, til at undersøge de umiddelbare konsekvenser af en given paradigme for visuel perception. Det forbinder også elektrofysiologiske og histologiske undersøgelser med funktionel v...

Baseret på sin gunstige neuro-anatomiske struktur gnaver visuelle system giver enestående muligheder for at vurdere farmakologiske forbindelser og deres evne til at mægle neuroprotection 1 eller pro-regenererende effekter 2,3. Desuden giver det undersøgelser af de funktionelle og neuro-anatomiske karakteristika af mus mutanter, som for nylig eksemplificeret for mus mangler den præsynaptiske stilladser protein Fagot 4.. Desuden et bredt spektrum af supplerende værktøjer giver ekstra featuring af retinal ganglion celle (RGC) og RGC Axon-numre samt RGC aktivitet, f.eks, ved elektroretinografi og adfærdsmæssige test, samt bestemmelse af kortikale omrokeringer af optisk billeddannelse af iboende signaler. Den seneste tekniske udvikling i laser mikroskopi aktivere in situ visualisering af RGC regenerering af dyb væv fluorescens billeddannelse i hele mount eksemplarer af synsnerven (ON) og hjerne. I denne histologiCal tilgang, tetrahydrofuran baseret væv clearing i kombination med lys ark fluorescensmikroskopi tillader opløsning af enkelte fibre, genindtræde i deafferented ON og optisk tarmkanalen 5.. Mens sådanne teknikker kan være overlegen i opløsning og bestemmelse af vækstmønstre, de ikke gør det muligt gentagne og langsgående analyser af individuelle vækst begivenheder, som er særligt ønskede at vurdere processen for langsigtede regenerering. Kontrast-forstærket MRI har været ansat i den minimale invasive visualisering af Retino-tectal projektion i mus og rotter 6,7. Dette kan opnås ved direkte intraokulært levering af paramagnetiske ioner (fx Mn2 +) til retinale celler. Som et calcium analog, er Mn 2 + indarbejdet i RGC somata via spændingsafhængige calcium kanaler og aktivt transporteres langs axonale cytoskeleton af det intakte ON og optisk tarmkanalen. Mens det ophobes i hjernekerneraf visuel projektion, dvs laterale geniculate nucleus (LGN) og overlegen colliculus (SC), transsynaptisk formering ind i den primære visuelle cortex vises ubetydelig 8,9, selv om det kan forekomme 10,11. Under MR sekventering, paramagnetiske Mn 2 + øger MR kontrast hovedsageligt ved at forkorte T 1 spin-gitter afslapning tid 12. Sådanne Mn 2 + forstærket MRI (MEMRI) er blevet anvendt med succes i forskellige neuro-anatomiske og funktionelle studier af rotter, herunder vurdering af axonal regenerering og degeneration efter ON skade 13,14, den præcise anatomiske kortlægning af Retino-tectal projektion 15 samt bestemmelse af axonale transport egenskaber efter farmakologisk behandling 16. Nylige forbedringer i dosering, toksicitet og kinetik af neuronale Mn 2 + optagelse og transport, samt forbedrede MR-protokoller har udvidet sin ansøgning til undersøgelser af transgenemus 9 ved hjælp af 3 Tesla scannere, der almindeligvis anvendes i klinisk praksis 17.. Her præsenterer vi en MEMRI protokol egnet til langsgående in vivo billeddannelse af muse Retino-tectal projektion og eksemplificere dets anvendelighed ved at vurdere Mn 2 + afhængig signal enhancement under naive og forskellige neurodegeneration forhold. Vores protokol lægger særlig vægt på erhvervelse MR data i et moderat 3 T magnetfelt, der generelt er mere tilgængeligt end dedikerede dyre scannere. I naive mus, vi illustrere, hvordan tarmkanalen-specifikke signal intensitet kan være væsentligt og reproducerbart blive forøget efter intravitreal (ivit) Mn 2 + ansøgning. Kvantitativt, Mn 2 + udbredelse langs den visuelle projektion sker uafhængigt af den normale aldringsproces (målt mellem 3 og 26 måneder gamle mus) og augmentation er refraktær over for visuel stimulation og tilpasning til mørket. I modsætning hertil, Mn <sop&gt; 2 + berigelse i thalamus og midthjernen centre er formindsket efter akut ON knusningsskade 18 samt i nfkb1 knock-out mus (P50 KO), der lider af spontan apoptotisk RGC død og ON degeneration 19.. Således i udvidelse til konventionel histologisk analyse, langsgående MEMRI analyse af de enkelte dyr muliggør profilering af unikke kinetik af neurodegenerative processer. Dette skulle vise sig nyttigt for undersøgelser af neurobeskyttelse og axonal regenerering er forbundet med farmakologiske eller genetiske indgreb.

<table border="0" cellpadding="0" cellspacing="0" width="934">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:363px;">Manganese (II) chloride solution 1M</td>
			<td style="width:269px;">Sigma Aldrich, Taufkirchen, Germany</td>
			<td style="width:119px;">M1787</td>
			<td style="width:184px;">MEMRI contrast reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;"></td>
			<td></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Conjuncain</td>
			<td>Dr. Mann Pharma, Berlin, Germany</td>
			<td style="width:119px;">PZN 7617666</td>
			<td>0.4% oxybuprocaine hydrochloride</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Floxal eye drops</td>
			<td>Dr. Mann Pharma, Berlin, Germany</td>
			<td style="width:119px;">PZN 3820927</td>
			<td>3 mg/ml ofloxacin</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ointment panthenol</td>
			<td>Jenapharm, Jena, Germany</td>
			<td style="width:119px;">PZN 3524531</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Chloral hydrate&#160;</td>
			<td style="width:269px;">Sigma Aldrich, Taufkirchen, Germany</td>
			<td style="width:119px;">C8383</td>
			<td>420-450 mg/kg body weight</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Actavis, Munich, Germany</td>
			<td style="width:119px;">PZN 7253744</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;"></td>
			<td></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hamilton syringe&#160;</td>
			<td style="width:269px;">Hamilton Company, Reno, NV, USA</td>
			<td style="width:119px;">7634-01</td>
			<td>SYR 5 &#181;l, 75 RN, no NDL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">34 G&#160; needle (34/35/pst4/tapN)</td>
			<td style="width:269px;">Hamilton Company, Reno, NV, USA</td>
			<td style="width:119px;">207434/00</td>
			<td>removable needle RN, 34 gauge, lenght 38.1 mm, point style 4</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Binocular Stemi-2000</td>
			<td style="width:269px;">Zeiss, Oberkochen, Germany</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:363px;"></td>
			<td style="width:269px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3T MRI scanner Magnetom TIM Trio</td>
			<td colspan="2">Siemens Medical Solutions, Erlangen, Germany</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rat head coil</td>
			<td>Doty Scientific Inc., Columbia, SC, USA</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mouse holder</td>
			<td style="width:269px;"></td>
			<td style="width:119px;"></td>
			<td>custom made</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Red light lamp</td>
			<td style="width:269px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;"></td>
			<td style="width:269px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:363px;">Frozen section medium NEG-50</td>
			<td style="width:269px;">Thermo Fisher Scientific, Schwerte, Germany</td>
			<td style="width:119px;">6502</td>
			<td>tissue embedding for cryo-sections</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;"></td>
			<td style="width:269px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:363px;">Sodium dihydrogen phosphate monohydrate (NaH2PO4)</td>
			<td>Merck, Darmstadt, Germany</td>
			<td style="width:119px;">106346</td>
			<td rowspan="2" style="width:184px;">for sulfide perfusion&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:363px;">Sodium sulfide nonahydrate (Na2S &#215; 9 H2O)</td>
			<td>Sigma Aldrich, Taufkirchen, Germany</td>
			<td style="width:119px;">208043</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:363px;">gum arabic</td>
			<td style="width:269px;">Roth, Arlesheim, Switzerland</td>
			<td style="width:119px;">4159</td>
			<td rowspan="5">for TIMM staining</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:363px;">Hydroquinone (C6H6O2)</td>
			<td style="width:269px;">Roth, Arlesheim, Switzerland</td>
			<td style="width:119px;">3586</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:363px;">Citric acid (C6H8O7)</td>
			<td style="width:269px;">Roth, Arlesheim, Switzerland</td>
			<td style="width:119px;">6490</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:363px;">Tri-sodium citrate dihydrate (C6H5Na3O7 x 2H2O)</td>
			<td style="width:269px;">Merck, Darmstadt, Germany</td>
			<td style="width:119px;">106448</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:363px;">Silver nitrate (AgNO3)</td>
			<td style="width:269px;">Roth, Arlesheim, Switzerland</td>
			<td style="width:119px;">7908</td>
		</tr>
	</tbody>
</table>

in vivo imaging of optic nerve fiber integrity by contrast-enhanced mri in mice

Det gnaver visuelle system omfatter retina-ganglieceller og deres axoner, der danner synsnerven at indtaste thalamiske og midthjernen centre, og postsynaptiske projektioner til den visuelle cortex. Baseret på sin særegne anatomiske struktur og praktisk tilgængelighed, er det blevet den foretrukne struktur for undersøgelser af neuronal overlevelse, axonal regeneration, og synaptisk plasticitet. Nylige fremskridt i MR-scanning har aktiveret in vivo visualisering af Retino-tectal del af denne fremskrivning ved hjælp af mangan medieret kontrast enhancement (MEMRI). Her præsenterer vi en MEMRI protokol til illustration af den visuelle projektion i mus, hvorved resolutioner (200 pm) 3 kan opnås ved hjælp af fælles 3 Tesla scannere. Vi viser, hvordan intravitreal injektion af en enkelt dosis på 15 nmol MnCl2 fører til en mættet forbedring af det intakte fremspring inden for 24 timer. Med undtagelse af nethinden, indregnes ændringer i signal intensitet uafdent af sammenfaldende visuel stimulation eller fysiologisk aldring. Vi mener desuden anvende denne teknik til langsgående overvåge aksonal degeneration i respons på akut synsnerven skade, et paradigme, som Mn 2 + transport helt anholdelser på læsion site. Omvendt aktiv Mn 2 + transport er kvantitativt i forhold til levedygtighed, nummer og elektriske aktivitet i Axon fibre. For en sådan analyse, vi eksemplificere Mn 2 + transport kinetik langs den visuelle vej i en transgen musemodel (NF-KB-p50 KO) viser spontan atrofi af sensoriske, herunder visuelle projektioner. I disse mus MEMRI indikerer, reduceret, men ikke forsinket Mn 2 + transport i forhold til vildtype-mus, og dermed afslører tegn på strukturelle og / eller funktionelle svækkelser af NF-KB-mutationer. Sammenfattende MEMRI bekvemt bro in vivo-analyser og efter slagtning histologi for characterizatipå nerve fiber integritet og aktivitet. Det er meget nyttigt for longitudinelle studier på axonal degeneration og regeneration, og undersøgelser af mutant mus for ægte eller inducerbare fænotyper.

Evne denne billeddannelse teknik til præcist at vurdere den vitalitet og funktionaliteten af den visuelle projektion afhængig præcise anvendelse af en ugiftig Mn 2 + dosering til glaslegeme og dets optagelse i rGCS. Denne store antagelse er testet i figur 1, hvor lag specifik Mn2 +-optagelse er påvist ved autometallografi (TIMM farvning) 21. Retina snit blev analyseret ved 24 timer efter ivit anvendelse af enten 15 nmol eller 150 nmol Mn2 +, eller...

Watch this Scientific Journal Video about In vivo Imaging of Optic Nerve Fiber Integrity by Contrast-Enhanced MRI in Mice at JoVE.com

In vivo Imaging of Optic Nerve Fiber Integrity by Contrast-Enhanced MRI in Mice

Denne video illustrerer en metode, ved hjælp af en klinisk 3 T scanner til kontrast-forstærkede MR scanning af det naive mus visuel projektion og til gentagne og langsgående in vivo studier af synsnerven degeneration er forbundet med akut synsnerven crush skade og kronisk synsnerven degeneration i knock-out mus (p50 KO).

In vivo billeddannelse af Optic Nerve Fiber Integritet ved kontrast-forstærket MRI i mus

The rodent visual system encompasses retinal ganglion cells and their axons that form the optic nerve to enter thalamic and midbrain centers, and ...

Research

JoVE Journal

Neuroscience

In vivo Imaging of Optic Nerve Fiber Integrity by Contrast-Enhanced MRI in Mice

Methods for Experimental Manipulations after Optic Nerve Transection in the Mammalian CNS

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 ...

Transfection of Mouse Retinal Ganglion Cells by in vivo Electroporation

Transfection of Mouse Retinal Ganglion Cells by in vivo Electroporation

The targeting and refinement of RGC projections to the midbrain is a popular and powerful model system for studying how precise patterns of neural ...

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

Funktionel Neuroimaging Med Ultrasonic blod-hjerne barrieren forstyrrelser og mangan-forstærket MRI

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

Fiber-optic Implantation for Chronic Optogenetic Stimulation of Brain Tissue

Fiberoptisk implantation for kronisk Optogenetic Stimulation af hjernevæv

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard ...

High-resolution In Vivo Manual Segmentation Protocol for Human Hippocampal Subfields Using 3T Magnetic Resonance Imaging

High-resolution In Vivo Manual Segmentation Protocol for Human Hippocampal Subfields Using 3T Magnetic Resonance Imaging

Høj opløsning<em&gt; In vivo</em&gt; Manuel Segmentering Protokol for humant hippocampus Underfelter Brug 3T Magnetic Resonance Imaging

The human hippocampus has been broadly studied in the context of memory and normal brain function and its role in different neuropsychiatric disorders has ...

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

Samtidige<em&gt; Ex vivo</em&gt; Functional Test af to nethinder af<em&gt; In vivo</em&gt; Elektroretinogrammet System

An In vivo electroretinogram (ERG) signal is composed of several overlapping components originating from different retinal cell types, as well as noise ...

Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination

Visuelt fremkaldt Potentiel optagelse i en rotte model for Eksperimentel Optic Nerve Demyelinisering

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to ...

High-resolution Structural Magnetic Resonance Imaging of the Human Subcortex In Vivo and Postmortem

High-resolution Structural Magnetic Resonance Imaging of the Human Subcortex In Vivo and Postmortem

Høj opløsning Strukturel Magnetic Resonance Imaging af Human subcortex<I&gt; In vivo</I&gt; Og Postmortem

The focus of this study was to test the resolution limits of structural MRI of a postmortem brain compared to living human brains. The resolution of ...

In Vivo Gene Transfer to Schwann Cells in the Rodent Sciatic Nerve by Electroporation

In Vivo Gene Transfer to Schwann Cells in the Rodent Sciatic Nerve by Electroporation

In vivo genoverførsel til Schwann-celler i Rodent iskiasnerven ved elektroporering

The formation of the myelin sheath by Schwann cells (SCs) is essential for rapid conduction of nerve impulses along axons in the peripheral nervous ...

In Vivo Two-photon Imaging of Cortical Neurons in Neonatal Mice

In Vivo Two-photon Imaging of Cortical Neurons in Neonatal Mice

In Vivo To-foton billeddannelse af kortikale neuroner i Neonatal mus

Two-photon imaging is a powerful tool for the in vivo analysis of neuronal circuits in the mammalian brain. However, a limited number of in vivo imaging ...