Name: methylnitrosourea (mnu)-induced retinal degeneration and regeneration in the zebrafish: histological and functional characteristics
Uploaded: 2014-10-20T14:00:00
Description: Watch this Scientific Journal Video about Methylnitrosourea MNU-induced Retinal Degeneration and Regeneration in the Zebrafish: Histological and Functional Characteristics at JoVE.com

We thank Monika Kilchenmann, Federica Bisignani and Agathe Duda for their excellent technical assistance.

Alle eksperimenter klæbet til erklæring til anvendelse af dyr i øjen og Vision Research for Foreningen for Forskning i Vision og Oftalmologi (ARVO). 1. Dyr <ol><li> Oprethold vildtype-zebrafisk (Danio rerio) af AB (Oregon) stamme i alderen 6-12 måneder under standardbetingelser i vand med en temperatur på 26,5 ° C og en 14/10 timers lys / mørke-cyklus 15. </li><li> Følg retningslinjerne dyr pleje af de involverede institutioner for dyreforsøg efter godkendelsen...

<ol>
	<li>Haddad, S., Chen, C. A., Santangelo, S. L., Seddon, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetics+of+age-related+macular+degeneration:+a+review+of+progress+to+date.">The genetics of age-related macular degeneration: a review of progress to date.</a> Surv. Ophthalmol. 51 (4), 316-363 (2006).</li><li>Bhatti, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinitis+pigmentosa,+pigmentary+retinopathies,+and+neurologic+diseases.">Retinitis pigmentosa, pigmentary retinopathies, and neurologic diseases.</a> Curr. Neurol. Neurosci. Rep. 6 (5), 403-413 (2006).</li><li>Hartong, D. T., Berson, E. L., Dryja, T. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinitis+pigmentosa.">Retinitis pigmentosa.</a> Lancet. 368, 1795-1809 (2006).</li><li>Tsubura, A., Yoshizawa, K., Kuwata, M., Uehara, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+for+retinitis+pigmentosa+induced+by+MNU;+disease+progression,+mechanisms+and+therapeutic+trials.">Animal models for retinitis pigmentosa induced by MNU; disease progression, mechanisms and therapeutic trials.</a> Histol. Histopathol. 25, 233-248 (2010).</li><li>Machida, K., Urano, K., Yoshimura, M., Tsutsumi, H., Nomura, T., Usui, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Carcinogenic+comparative+study+on+rasH2+mice+produced+by+two+breeding+facilities.">Carcinogenic comparative study on rasH2 mice produced by two breeding facilities.</a> J. Toxicol. Sci. 33, 493-501 (2008).</li><li>Morton, 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=N-Methyl-N-Nitrosourea+(MNU):+A+positive+control+chemical+for+p53+/-+mouse+carcinogenicity+studies.">N-Methyl-N-Nitrosourea (MNU): A positive control chemical for p53+/- mouse carcinogenicity studies.</a> Toxicol. Pathol. 36, 926-931 (2008).</li><li>Terracini, B., Testa, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Carcinogenicity+of+a+single+administration+of+N-nitrosomethylurea:+a+comparison+between+newborn+and+5-week-old+mice+and+rats.">Carcinogenicity of a single administration of N-nitrosomethylurea: a comparison between newborn and 5-week-old mice and rats.</a> 24, 588-598 (1970).</li><li>Zulliger, R., Lecaud&#233;, S., Eigeldinger-Berthou, S., Wolf-Schnurrbusch, U. E. K., Enzmann, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Caspase-3-independent+photoreceptor+degeneration+by+N-methyl-N-nitrosourea+(MNU)+induces+morphological+and+functional+changes+in+the+mouse+retina.">Caspase-3-independent photoreceptor degeneration by N-methyl-N-nitrosourea (MNU) induces morphological and functional changes in the mouse retina.</a> Graefes Arch. Clin. Exp. Ophthalmol. 249, 859-869 (2011).</li><li>Tappeiner, 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=Characteristics+of+rod+regeneration+in+a+novel+zebrafish+retinal+degeneration+model+using+N-methyl-N-nitrosourea+(MNU).">Characteristics of rod regeneration in a novel zebrafish retinal degeneration model using N-methyl-N-nitrosourea (MNU).</a> PLOS One. 12, (2013).</li><li>Bilotta, J., Saszik, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+zebrafish+as+a+model+visual+system.">The zebrafish as a model visual system.</a> Int. J. Dev. Neurosci. 19, 621-629 (2001).</li><li>Fleisch, C., Neuhauss, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visual+Behavior+in+Zebrafish.">Visual Behavior in Zebrafish.</a> Zebrafish. 3, 191-201 (2006).</li><li>Hodel, C., Neuhauss, S. C. F., Biehmaier, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+course+and+development+of+light+adaptation+processes+in+the+outer+zebrafish+retina.">Time course and development of light adaptation processes in the outer zebrafish retina.</a> InterScience. 288, 653-662 (2006).</li><li>Gemberling, M., Bailey, T. J., Hyde, D. R., Poss, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+zebrafish+as+a+model+for+complex+tissue+regeneration.">The zebrafish as a model for complex tissue regeneration.</a> Trends Genet. 29 (11), 611-620 (2013).</li><li>Brockerhoff, S. E., Fadool, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetics+of+photoreceptor+degeneration+and+regeneration+in+zebrafish.">Genetics of photoreceptor degeneration and regeneration in zebrafish.</a> Cell Mol. Life Sci. 68, 651-659 (2011).</li><li>Brand, M., Granato, M., N&#252;sslein-Volhard, C., N&#252;sslein-Volhard, C., Dahm, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Keeping+and+raising+zebrafish.">Keeping and raising zebrafish.</a> Zebrafish: A Practical Approach. , 7-38 (2002).</li><li>Tappeiner, C., Gerber, S., Enzmann, V., Balmer, J., Jazwinska, A., Tschopp, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visual+acuity+and+contrast+sensitivity+of+adult+zebrafish.">Visual acuity and contrast sensitivity of adult zebrafish.</a> Front. Zool. 9 (1), 10 (2012).</li><li>Bailey, T. J., Fossum, S. L., Fimbel, S. M., Montgomery, J. E., Hyde, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+inhibitor+of+phagocytosis,+O-phospho-L-serine,+suppresses+M&#252;ller+glia+proliferation+and+cone+cell+regeneration+in+the+light-damaged+zebrafish+retina.">The inhibitor of phagocytosis, O-phospho-L-serine, suppresses M&#252;ller glia proliferation and cone cell regeneration in the light-damaged zebrafish retina.</a> Exp. Eye Res. 91, 601-612 (2010).</li><li>Nelson, C. M., Hyde, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=M&#252;ller+glia+as+a+source+of+neuronal+progenitor+cells+to+regenerate+the+damaged+zebrafish+retina.">M&#252;ller glia as a source of neuronal progenitor cells to regenerate the damaged zebrafish retina.</a> Adv. Exp. Med. Biol. 723, 425-430 (2012).</li><li>Prusky, G. T., Alam, N. M., Beekman, S., Douglas, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+quantification+of+adult+and+developing+mouse+spatial+vision+using+a+virtual+optomotor+system.">Rapid quantification of adult and developing mouse spatial vision using a virtual optomotor system.</a> Invest. Ophthalmol Vis. Sci. 45 (12), 4611-4616 (2004).</li><li>Beck, J. C., Gilland, E., Tank, D. W., Baker, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantifying+the+ontogeny+of+optokinetic+and+vestibulo-ocular+behaviors+in+zebrafish,+medaka,+and+goldfish.">Quantifying the ontogeny of optokinetic and vestibulo-ocular behaviors in zebrafish, medaka, and goldfish.</a> J. Neurophysiol. 92 (6), 3546-3561 (2004).</li><li>Fimbel, S. M., Montgomery, J. E., Burket, C. T., Hyde, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regeneration+of+Inner+Retinal+Neurons+after+Intravitreal+Injection+of+Ouabain+in.">Regeneration of Inner Retinal Neurons after Intravitreal Injection of Ouabain in.</a> Zebrafish. J. Neurosci. 27, 1712-1724 (2007).</li><li>Sherpa, 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=Ganglion+cell+regeneration+following+whole-retina+destruction+in+zebrafish.">Ganglion cell regeneration following whole-retina destruction in zebrafish.</a> Dev. Neurobiol. 68, 166-181 (2008).</li><li>Yurco, P., Cameron, 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=Responses+of+M&#252;ller+glia+to+retinal+injury+in+adult+zebrafish.">Responses of M&#252;ller glia to retinal injury in adult zebrafish.</a> Vision Res. 45, 991-1002 (2005).</li></ol>

Tidligere har vores gruppe overført MNU model fotoreceptordegenerering fra gnavere i zebrafisk systemet 9. De efterfølgende hændelser blev beskrevet og fulgt i op til 30 dage. I denne periode opstod fuldstændig retina morfologiske degeneration og regeneration efter den indledende behandling. Først histologi afslører en reduceret stang celletal fra dag 3 på med et minimum på dag 8. Tilsvarende TUNEL farvning identificerer apoptose af stangen fotoreceptorer 3 dage efter MNU behandling. Regenerering begy...

Vision er den mest afgørende mening for mennesket og dets leverfunktion har en stor samfundsøkonomisk virkning. I den udviklede verden, retinale degenerative sygdomme er den hyppigste årsag til synstab og blindhed blandt ældre voksne 1. Årsagen til de fleste degenerative retinale sygdomme er kun delvist forstået og terapeutiske løsninger til at genvinde tabte vision er meget begrænsede. Retinitis pigmentosa er et typisk eksempel på en retina degenerativ sygdom med primær fotoreceptor tab 2-3. N-methyl-N -nitrosourea (MNU) inducerer retinal degeneration og er derfor almindeligt brugt i gnavere at modellere sygdomme med primær fotoreceptor celledød 4. Det er et alkyleringsmiddel og fører til benigne og maligne tumorer, som normalt vises flere måneder efter eksponering 5-7. Desuden bevirker specifikke fotoreceptor celledød inden for en kort sigt observationsperiode. Derved tabet af retinal lag struktre og betydelig retinal udtynding blev observeret i en koncentrationsafhængig måde. Retina gliaceller blev aktiveret, men ingen ændringer i det retinale pigment epitel (RPE) findes. Endoplasmatiske reticulum (ER) stressrelateret apoptose synes at være den vigtigste vej for MNU aktion i nethinden 8. Vi har for nylig indført MNU som en kemisk model til at inducere fotoreceptordegenerering i zebrafisk 9. Blandt andre grunde har zebrafisk (Danio rerio) blevet vigtigt i visuel forskning grund af lighederne i dens visuelle system til andre hvirveldyr 10. Den ydre nethinde indeholder fotoreceptorer, som kan inddeles i fire forskellige kegle typer med peak følsomhedsberegninger i det ultraviolette, kort, mellemlang og lang bølgelængde af det synlige spektre og en stang fotoreceptor type. I den indre nukleare lag (INL) er cellelegemerne bipolære, vandret og amacrine interneuroner fundet, som welll som celle soma af Muller gliaceller. I det ydre netformige lag (OPL) dannes synaptiske kontakter mellem fotoreceptorer og den indre nethinde, mens cellelag nærmest linsen er ganglion cellelag (GC), hvilke komponenter danner lange axoner omfatter synsnerven og optisk tarmkanalen . Synaptiske kontakter mellem ganglieceller og cellerne i det indre kernelag er dannet i det indre netformige lag (IPL) 11. RPE ligger udenfor neurosensoriske nethinde og omgiver fotoreceptor ydre segmenter med lange apikale microvilli 12. Endvidere zebrafisk er meget regenerative og i stand til at regrow læderede hjerne, nethinde, rygmarv, hjerte og andre væv 13. Når beskadigelse af nethinden opstår, celler i INL, der formodes at være Müller celler aktiveres og har potentialet til at differentiere til forskellige retinale celletyper. Desuden er de også generere stang stamceller, der er placeret i ONL. Et andet såurce der forsyner nethinden af ​​voksne zebrafisk med nye celler er ciliaere randzone. Denne kilde er nødvendig for at opnå en konstant tæthed af stangen fotoreceptorer i de stadigt voksende zebrafisk øje 14. Den MNU model kan anvendes som en simpel og reproducerbar degenerering / regenerering fremgangsmåde for retinavæv. På grund af visse ligheder mellem biologiske processer i zebrafisk og i mennesker dette kunne åbne dørene for at identificere de involverede celledødsveje og at screene potentielle neurobeskyttende lægemidler. Baseret på en tidligere undersøgelse fra vores gruppe, vi nu illustrere metoderne for denne MNU-induceret zebrafisk model af retinal de- og deraf følgende regenerering, herunder funktionelle ændringer med ifølge laboratorium videoer 9.

<table border="0" cellpadding="0" cellspacing="0" >
	<tbody>
		<tr >
			<td >Acetic acid</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>A6283</td>
			<td></td>
		</tr>
		<tr >
			<td >Ammonia</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>294993</td>
			<td>0.80%</td>
		</tr>
		<tr >
			<td >Bovine serum albumine (BSA)</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>05470</td>
			<td></td>
		</tr>
		<tr >
			<td >Dako Pen</td>
			<td>Dako, Glostrup, Danmark</td>
			<td>S2002</td>
			<td></td>
		</tr>
		<tr >
			<td >DAPI mounting medium</td>
			<td>Vector Labs, Burlingame, CA, USA</td>
			<td>H-1200</td>
			<td></td>
		</tr>
		<tr >
			<td >Eosin</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>45260</td>
			<td></td>
		</tr>
		<tr >
			<td >Ethanol</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>2860</td>
			<td>100%, 96%, 70%</td>
		</tr>
		<tr >
			<td >Ethylenediaminetetraacetic acid (EDTA)</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>ED</td>
			<td></td>
		</tr>
		<tr >
			<td >Ethyl 3-aminobenzoate methanesulfonate salt</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>E10521</td>
			<td>Tricaine</td>
		</tr>
		<tr >
			<td >Eukitt</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>3989</td>
			<td></td>
		</tr>
		<tr >
			<td >Goat anti-rabbit Alexa 594</td>
			<td>Life Technologies, Zug, Switzerland</td>
			<td>A11012</td>
			<td></td>
		</tr>
		<tr >
			<td >Goat normal serum</td>
			<td>Dako, Glostrup, Danmark</td>
			<td>X0907</td>
			<td></td>
		</tr>
		<tr >
			<td >Hydrochloric acid</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>320331</td>
			<td>0.20%</td>
		</tr>
		<tr >
			<td >In situ Cell Death Detection Kit</td>
			<td>Roche Applied Sciences, Rotkreuz, Switzerland</td>
			<td>11684795910</td>
			<td>TUNEL Kit</td>
		</tr>
		<tr >
			<td >Mayer&#39;s hemalum solution</td>
			<td>Merck, Darmstadt, Germany</td>
			<td>109249</td>
			<td></td>
		</tr>
		<tr >
			<td >Methylnitrosourea (MNU)</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>N4766</td>
			<td>Toxic</td>
		</tr>
		<tr >
			<td >OptoMotry</td>
			<td>CerebralMechanics, Lethbridge, AB, Canada</td>
			<td>n.a.</td>
			<td></td>
		</tr>
		<tr >
			<td >Paraformaldehyde (PFA)</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>P6148</td>
			<td></td>
		</tr>
		<tr >
			<td >Phosphate buffered saline (PBS)</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>P5368</td>
			<td></td>
		</tr>
		<tr >
			<td >Proteinase K</td>
			<td>Dako, Glostrup, Danmark</td>
			<td>S3004</td>
			<td></td>
		</tr>
		<tr >
			<td >Rabbit anti-PCNA</td>
			<td>Santa Cruz Biotechnology, Santa Cruz, USA</td>
			<td>sc-33756</td>
			<td></td>
		</tr>
		<tr >
			<td >Superfrost Plus glass slides</td>
			<td>Gehard Menzel GmbH, Braunschweig, Germany</td>
			<td>10149870</td>
			<td></td>
		</tr>
		<tr >
			<td >Tris buffered saline (TBS)</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>P5912</td>
			<td></td>
		</tr>
		<tr >
			<td >Trizma&#210; base</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>T1503</td>
			<td></td>
		</tr>
		<tr >
			<td >Tween 20</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>P1379</td>
			<td></td>
		</tr>
		<tr >
			<td >Xylene</td>
			<td>Sigma-Aldrich, Buchs, Switzerland</td>
			<td>534056</td>
			<td></td>
		</tr>
		<tr >
			<td >Zebrafish (Danio rerio) AB (Oregon) strain</td>
			<td>University of Fribourg, Dept. of Biology</td>
			<td>n.a.</td>
			<td>Own fish facility</td>
		</tr>
	</tbody>
</table>

methylnitrosourea (mnu)-induced retinal degeneration and regeneration in the zebrafish: histological and functional characteristics

Retinale degenerative sygdomme, fx retinitis pigmentosa, med deraf følgende fotoreceptor skader tegner sig for størstedelen af synstab i den industrielle verden. Dyremodeller af central betydning for at studere sådanne sygdomme. I denne forbindelse fotoreceptoren-specifikke toksin N-methyl-N -nitrosourea (MNU) er blevet meget udbredt i gnavere til farmakologisk fremkalde nethindedegeneration. Tidligere har vi etableret en MNU-induceret nethindedegeneration model i zebrafisk, en anden populær model-system i visuel forskning. En fascinerende forskel til pattedyr er den vedvarende neurogenese i den voksne zebrafisk nethinden og dens regenerering efter skader. For at kvantificere denne observation har vi ansat visuelle skarphed målinger i den voksne zebrafisk. Derved blev den optokinetic refleks anvendes til at følge funktionelle ændringer i ikke-bedøvede fisk. Dette blev suppleret med histologi samt immunhistokemiske staining til apoptose (TUNEL) og proliferation (PCNA) at korrelere udviklingslandene morfologiske ændringer. Sammenfattende forekommer apoptose af fotoreceptorer tre dage efter MNU behandling, som er efterfulgt af en markant reduktion af celler i det ydre kernelag (ONL). Derefter observeres proliferation af celler i det indre kernelag (INL) og ONL. Heri viser vi, at ikke kun en fuldstændig histologisk, men også en funktionel regenerering sker over et tidsforløb på 30 dage. Nu skal vi illustrere de metoder, til at kvantificere og følge op zebrafisk retinal de- og regeneration ved hjælp af MNU i en video-format.

Synsstyrke: Den eksperimentelle opstilling [rumlige frekvens: 0,042 cirkler / grad (c / d); kontrast: 100%; afdrift hastighed: 20 grader / sekund (d / SRC); baggrundslys luminans: 152 cd / m 2] i denne undersøgelse aktiveret OKR vurdering af den voksne zebrafisk. Den gennemsnitlige varighed af VA måling var omkring 5 - 10 minutter til hver zebrafisk, som tolereres proceduren godt. Synsstyrke før MNU eksponering var 0,577 ± 0,014 cyklusser / grad (c / d). Figur 1</stron...

Watch this Scientific Journal Video about Methylnitrosourea MNU-induced Retinal Degeneration and Regeneration in the Zebrafish: Histological and Functional Characteristics at JoVE.com

Methylnitrosourea MNU-induced Retinal Degeneration and Regeneration in the Zebrafish: Histological and Functional Characteristics

Heri vi demonstrere kvantificering af retinal de- og regenerering og dens indvirkning på visuel funktion ved anvendelse af N-methyl-N -nitrosourea i den voksne zebrafisk. Tab af synsstyrke og nedsat fotoreceptor numre blev efterfulgt af proliferation i den indre nukleare lag. Komplet morfologiske og funktionelle regenerering forekom 30 dage efter den indledende behandling.

Methylnitrosourea (MNU) -inducerede nethindedegeneration og Regeneration i zebrafisk: Histologisk og funktionelle egenskaber

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Methylnitrosourea (MNU)-induced Retinal Degeneration and Regeneration in the Zebrafish: Histological and Functional Characteristics

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In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

In vivo Elektroporation af Morpholinos i den voksne zebrafisk Retina

Many devastating inherited eye diseases result in progressive and irreversible blindness because humans cannot regenerate dying or diseased retinal ...

In vivo Electroporation of Morpholinos into the Regenerating Adult Zebrafish Tail Fin

In vivo Electroporation of Morpholinos into the Regenerating Adult Zebrafish Tail Fin

In vivo elektroporation af Morpholinos i Regenerating Adult zebrafisk Tail Fin

Certain species of urodeles and teleost fish can regenerate their tissues. Zebrafish have become a widely used model to study the spontaneous regeneration ...

Gavaging Adult Zebrafish

Gavaging Voksen Zebrafisk

The zebrafish has become an important in vivo model in biomedical research. Effective methods must be developed and utilized to deliver compounds or ...

An Assay for Lateral Line Regeneration in Adult Zebrafish

En analyse for sidelinje Regeneration i Voksen Zebrafisk

Due to the clinical importance of hearing and balance disorders in man, model organisms such as the zebrafish have been used to study lateral line ...

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

Funktionel Opløsning og kanalaktivitet Målinger af renset Vildtype og Mutant CFTR Protein

The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a unique channel-forming member of the ATP Binding Cassette (ABC) superfamily of ...

Gap Junctional Intercellular Communication: A Functional Biomarker to Assess Adverse Effects of Toxicants and Toxins, and Health Benefits of Natural Products

Gap junktion intercellulær kommunikation: En funktionel biomarkør at vurdere negative virkninger af giftstoffer og toksiner, og Sundhedsmæssige fordele af Naturprodukter

This protocol describes a scalpel loading-fluorescent dye transfer (SL-DT) technique that measures intercellular communication through gap junction ...

Immunolabelling Myofiber Degeneration in Muscle Biopsies

Immunolabelling myofiber degeneration i muskelbiopsier

The necrosis of muscle fibres (myonecrosis) plays a central role in the pathogenesis of several muscle conditions, including muscular dystrophies. ...

Optical Sectioning and Visualization of the Intervertebral Disc from Embryonic Development to Degeneration

Optisk sektion og visualisering af den intervertebrale disk fra embryonisk udvikling til degeneration

Intervertebral disc (IVD) degeneration is a leading cause of low back pain and it entails a high degree of impairment for the affected individuals. To ...