Name: longitudinal in vivo imaging of the cerebrovasculature: relevance to cns diseases
Uploaded: 2016-12-06T14:00:00
Description: Watch this Scientific Journal Video about Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases at JoVE.com

Forfatterne vil gerne anerkende Ligue Francaise contre l&#39;Epilepsie (til MA-L), Institut National de la Santé et de la Recherche Médicale Grant AVENIR R12087FS (til FJ), give fra universitetet i Montpellier (til FJ) og tilskud fra Federation pour la Recherche sur le Cerveau (til NM). Vi anerkender den tekniske bistand fra Chrystel Lafont på IPAM in vivo imaging kerneplatform facilitet i Montpellier. Vi takker også Mary Vernov (Weill Cornell Medical College) til korrekturlæsning manuskriptet.

Mus er tilladt ad libitum adgang til mad, vand og holdt i en 12-timers lys-mørke-cyklus. Alle procedurer, der involverer forsøgsdyr var i overensstemmelse med nationale og europæiske love og blev godkendt af det franske Ministerium for uddannelse og videnskabelig forskning (CEEA-LR-00.651-01). I alt 6 transgen 5xFAD og 4 kuldsøster vildtype (WT) kontrolmus blev anvendt til denne procedure. 1. Præoperativ forberedelse <ol><li> Intraperitonealt (IP) injicere Methoxy-x 04 (10 mg / kg) 48 timer før ...

<ol>
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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=Abeta+is+targeted+to+the+vasculature+in+a+mouse+model+of+hereditary+cerebral+hemorrhage+with+amyloidosis.">Abeta is targeted to the vasculature in a mouse model of hereditary cerebral hemorrhage with amyloidosis.</a> Nat Neurosci. 7 (9), 954-960 (2004).</li><li>Oakley, 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=Intraneuronal+beta-amyloid+aggregates,+neurodegeneration,+and+neuron+loss+in+transgenic+mice+with+five+familial+Alzheimer's+disease+mutations:+potential+factors+in+amyloid+plaque+formation.">Intraneuronal beta-amyloid aggregates, neurodegeneration, and neuron loss in transgenic mice with five familial Alzheimer's disease mutations: potential factors in amyloid plaque formation.</a> J Neurosci. 26 (40), 10129-10140 (2006).</li><li>Liston, 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=Circadian+glucocorticoid+oscillations+promote+learning-dependent+synapse+formation+and+maintenance.">Circadian glucocorticoid oscillations promote learning-dependent synapse formation and maintenance.</a> Nat Neurosci. 16 (6), 698-705 (2013).</li><li>Yang, G., Pan, F., Parkhurst, C. N., Grutzendler, J., Gan, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thinned-skull+cranial+window+technique+for+long-term+imaging+of+the+cortex+in+live+mice.">Thinned-skull cranial window technique for long-term imaging of the cortex in live mice.</a> Nat Protoc. 5 (2), 201-208 (2010).</li><li>Klunk, W. 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=Imaging+Abeta+plaques+in+living+transgenic+mice+with+multiphoton+microscopy+and+methoxy-X04,+a+systemically+administered+Congo+red+derivative.">Imaging Abeta plaques in living transgenic mice with multiphoton microscopy and methoxy-X04, a systemically administered Congo red derivative.</a> J Neuropathol Exp Neurol. 61 (9), 797-805 (2002).</li><li>Yardeni, T., Eckhaus, M., Morris, H. D., Huizing, M., Hoogstraten-Miller, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retro-orbital+injections+in+mice.">Retro-orbital injections in mice.</a> Lab Anim (NY). 40 (5), 155-160 (2011).</li><li>Cao, V. 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=In+vivo+two-photon+imaging+of+experience-dependent+molecular+changes+in+cortical+neurons.">In vivo two-photon imaging of experience-dependent molecular changes in cortical neurons.</a> J Vis Exp. 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Den åbne kraniet teknik til in vivo to-foton mikroskopi frembyder fordelen af ubegrænsede imaging sessioner af store billeddannelse felter 13,14. Men denne teknik også producerer betændelse i området af interesse 14, ofte uforenelige eller påvirker neuro-vaskulære læse-outs 15. Tværtimod har den indsnævrede kraniet transkraniel teknik ikke resultere i neuro-inflammation, således at en pålidelig billeddannelse af cerebrovaskulære strukturer og plakakkumulering 1...

Hjernen vaskulatur er en multi-cellulære struktur, der er anatomisk og funktionelt koblet til neuroner. En dynamisk omformning af fartøjer strækker sig over hele hjernens udvikling og under progressionen af patologier i det centrale nervesystem (CNS) 1,2. Det er almindeligt accepteret, at cerebrovaskulær skade er kendetegnende for flere CNS-sygdomme, herunder epilepsi, Alzheimers sygdom (AD), traumatisk hjerneskade og encephalitis 3,4. Derfor sporing cerebrovaskulære ændringer in vivo bliver mærkbar ved modellering CNS-sygdomme, fra påbegyndelse og ind kroniske faser. Som cerebrovaskulære ændringer forekommer ofte samtidig med neuronal skade eller plasticitet, billedbehandling af neuro-karsystem udgør en vigtig indgang til dechifrere CNS sygdom patofysiologi. Denne protokol beskriver en langsgående to-foton baseret procedure at spore ombygning af cerebrovasculature i en musemodel afAD, en progressiv patologi præget af cerebrovaskulære defekter på store og små kaliber fartøjer på grund af amyloidogen plak deposition 5-7. Denne procedure giver mulighed for visualisering af amyloidaflejringer og sporing af deres position og vækst i forhold til neurovaskulær remodeling hele forløbet af sygdommen. Vital fluorescerende farvestoffer injiceres før hver billeddannelse session for visualisering af cerebrovasculature og amyloide plaques i transgene AD mus 8. Gentagne imaging sessioner af en ROI gennem en fortyndet kranium transkraniel vinduet er ikke-invasiv og den foretrukne metode til at vurdere neurovaskulær remodeling i den levende musehjerne 2,5,9,10. Proceduren nedenfor skitserer den kirurgiske protokol, erhvervelse og behandling image. Den tidlige progression af cerebral amyloid angiopati (CAA) for det meste på store leptomeningeal og gennemtrængende arterioler er præget.

<table border="0" cellpadding="0" cellspacing="0" width="627">
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		<col />
		<col />
		<col />
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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">methoxy-X04</td>
			<td style="width:125px;">tocris</td>
			<td align="right" style="width:119px;">4920</td>
			<td style="width:167px;">use 10 mg/Kg</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">FITC-Dextran 70Kda</td>
			<td style="width:125px;">sigma</td>
			<td align="right" style="width:119px;">46945</td>
			<td>use 100 mg/Kg</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">gelfoam/Bloxang</td>
			<td style="width:125px;">Bausch and Lomb</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">micorsurgical blade</td>
			<td style="width:125px;">surgistar</td>
			<td align="right" style="width:119px;">6900</td>
			<td>must be sharp and not dented</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">povidone-iodine</td>
			<td style="width:125px;">betadine</td>
			<td style="width:119px;"></td>
			<td>antisceptic solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">binocular stereomicroscope</td>
			<td style="width:125px;">olympus</td>
			<td style="width:119px;">SX10</td>
			<td>optimal image contrast is crucial for this procedure</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">2-photon microscope</td>
			<td style="width:125px;">zeiss</td>
			<td style="width:119px;"></td>
			<td>Zeiss LSM 710mp</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">fine scissors-toughcut</td>
			<td style="width:125px;">Fine science tools</td>
			<td>14058-09</td>
			<td>this scissors are optimized for cutting skin and soft tissue</td>
		</tr>
	</tbody>
</table>

longitudinal in vivo imaging of the cerebrovasculature: relevance to cns diseases

Remodeling af hjernen vaskulaturen er en fælles træk af hjernen patologier. In vivo-billeddannelsesteknikker er fundamentale for påvisning cerebrovaskulær plasticitet eller skader overtid og i forhold til neuronal aktivitet eller blodstrømmen. In vivo to-foton mikroskopi tillader studiet af den strukturelle og funktionelle plasticitet af store cellulære enheder i den levende hjerne. Især den indsnævrede-kraniet vindue præparat tillader visualisering af corticale områder af interesse (ROI) uden at inducere signifikant hjernebetændelse. Gentagne imaging sessioner af kortikale ROI er mulige, giver karakterisering af sygdomstilstande kendetegnende over en periode i progression af talrige CNS-sygdomme. Denne teknik adgang til pial strukturer i 250 um af hjernen er afhængig af påvisning af fluorescerende prober kodes af genetiske cellulære markører og / eller vitale farvestoffer. Sidstnævnte (f.eks fluorescerende dextraner) anvendes til at kortlægge Luminal rum for cerebrovaskulære strukturer. Germane protokollen beskrevet heri er anvendelsen af en in vivo markør af amyloidaflejringer, methoxy-O4, at vurdere Alzheimers sygdom (AD) progression. Vi beskriver også den post-erhvervelse billedbehandling bruges til at spore vaskulære ændringer og amyloid aflejringer. Mens øjeblikket fokus på en model for AD, den beskrevne protokol er relevant for andre CNS-lidelser, hvor patologiske cerebrovaskulære forandringer forekomme.

Denne protokol beskriver en fremgangsmåde til visualisering af cerebrovasculature og amyloidaflejringer overtid. Fluorescerende farvestoffer blev injiceret at mærke amyloide aflejringer (methoxy-XO4) 11 og at fylde cerebrovaskulære lumen (FITC-dextran) 1. 3D billede analyse softwaremoduler blev brugt til at skabe 3D-billeder af en konstant synsfelt fanget på hinanden følgende tidspunkter. Repræsentative billeder opnået i somatosensoriske cortex af 5XFAD mus <...

Watch this Scientific Journal Video about Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases at JoVE.com

Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

Dette håndskrift beskrives en procedure til at spore ombygning af cerebrovasculature under amyloid plaque akkumulation in vivo ved anvendelse langsgående to-foton mikroskopi. En tyndet-kranium præparat muliggør visualisering af fluorescerende farvestoffer til at vurdere progressionen af ​​cerebrovaskulær skade i en musemodel for Alzheimers sygdom.

Longitudinal<em&gt; In vivo</em&gt; Imaging af Cerebrovasculature: Relevans til CNS-sygdomme

Remodeling of the brain vasculature is a common trait of brain pathologies. In vivo imaging techniques are fundamental to detect cerebrovascular ...

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Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

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