Name: electrophysiological recording in the brain of intact adult zebrafish
Uploaded: 2013-11-19T17:00:00
Description: Watch this Scientific Journal Video about Electrophysiological Recording in the Brain of Intact Adult Zebrafish at JoVE.com

Dette arbejde blev støttet af NIH / NINDS Grant R01NS070159 (til TMD, JDL og ATS).

Alle eksperimentelle procedurer blev udført i nøje overensstemmelse med National Institutes of Health Guide til Pleje og anvendelse af forsøgsdyr og fulgte protokol # A2011 09-003, som blev revideret, godkendt og overvåget af University of Georgia Institutional Animal Care og brug Udvalg. 1.. Opsætning af udstyr <ol><li> Perfusionssystem for kraniotomi Immobilisering af den voksne nødvendiggør en intubation systemet til at levere opløst oxygen på fisken. En...

<ol>
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A. <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+brain+in+research+and+teaching:+a+simple+in+vivo+and+in+vitro+model+for+the+study+of+spontaneous+neural+activity.">The zebrafish brain in research and teaching: a simple in vivo and in vitro model for the study of spontaneous neural activity.</a> Adv Physiol Educ. 35, 188-196 (2011).</li><li>Makhankov, Y. V., Rinner, O., Neuhauss, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+inexpensive+device+for+non-invasive+electroretinography+in+small+aquatic+vertebrates.">An inexpensive device for non-invasive electroretinography in small aquatic vertebrates.</a> J. Neurosci. Methods. 135, 205-210 (2004).</li><li>Brockerhoff, S. 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=A+behavioral+screen+for+isolating+zebrafish+mutants+with+visual+system+defects.">A behavioral screen for isolating zebrafish mutants with visual system defects.</a> Proc. Natl. Acad. Sci. U.S.A.. 92, 10545-10549 (1995).</li><li>Baraban, S. C., Taylor, M. R., Castro, P. A., Baier, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pentylenetetrazole+induced+changes+in+zebrafish+behavior,+neural+activity+and+c-fos+expression.">Pentylenetetrazole induced changes in zebrafish behavior, neural activity and c-fos expression.</a> Neuroscience. 131, 759-768 (2005).</li><li>Baraban, S. 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=A+large-scale+mutagenesis+screen+to+identify+seizure-resistant+zebrafish.">A large-scale mutagenesis screen to identify seizure-resistant zebrafish.</a> Epilepsia. 48, 1151-1157 (2007).</li><li>Maximino, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Serotonin and anxiety: Neuroanatomical, pharmacological and functional aspects. , (2012).</li><li>Bally-Cuif, L., Vernier, P., Perry, S. F., Ekker, M., Farrell, A. P., Brauner, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Fish Physiology: Zebrafish. 29, (2010).</li><li>Kaslin, J., Nystedt, J. M., Ostergard, M., Peitsaro, N., Panula, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+orexin/hypocretin+system+in+zebrafish+is+connected+to+the+aminergic+and+cholinergic+systems.">The orexin/hypocretin system in zebrafish is connected to the aminergic and cholinergic systems.</a> J. Neurosci. 24, 2678-2689 (2004).</li><li>McLean, D. L., Fetcho, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ontogeny+and+innervation+patterns+of+dopaminergic,+noradrenergic,+and+serotonergic+neurons+in+larval+zebrafish.">Ontogeny and innervation patterns of dopaminergic, noradrenergic, and serotonergic neurons in larval zebrafish.</a> J. Comp. Neurol. 480, 38-56 (2004).</li><li>Mueller, T., Vernier, P., Wullimann, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+adult+central+nervous+cholinergic+system+of+a+neurogenetic+model+animal,+the+zebrafish+Danio+rerio.">The adult central nervous cholinergic system of a neurogenetic model animal, the zebrafish Danio rerio.</a> Brain Res. 1011, 156-169 (2004).</li><li>Higashijima, S., Schaefer, M., Fetcho, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurotransmitter+properties+of+spinal+interneurons+in+embryonic+and+larval+zebrafish.">Neurotransmitter properties of spinal interneurons in embryonic and larval zebrafish.</a> J. Comp. Neurol. 480, 19-37 (1002).</li><li>Tao, L., Lauderdale, J. D., Sornborger, A. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+Functional+Connectivity+between+Neuronal+Ensembles+with+Larval+Zebrafish+Transgenic+for+a+Ratiometric+Calcium+Indicator.">Mapping Functional Connectivity between Neuronal Ensembles with Larval Zebrafish Transgenic for a Ratiometric Calcium Indicator.</a> Front Neural Circuits. 5, 2 (2011).</li><li>Fan, 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=New+statistical+methods+enhance+imaging+of+cameleon+fluorescence+resonance+energy+transfer+in+cultured+zebrafish+spinal+neurons.">New statistical methods enhance imaging of cameleon fluorescence resonance energy transfer in cultured zebrafish spinal neurons.</a> J Biomed Opt. 12, 034017 (2007).</li><li>Burgess, H. A., Granato, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensorimotor+gating+in+larval+zebrafish.">Sensorimotor gating in larval zebrafish.</a> J. Neurosci. 27, 4984-4994 (2007).</li><li>Burgess, H. A., Schoch, H., Granato, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+retinal+pathways+drive+spatial+orientation+behaviors+in+zebrafish+navigation.">Distinct retinal pathways drive spatial orientation behaviors in zebrafish navigation.</a> Curr. Biol. 20, 381-386 (2010).</li><li>Mueller, K. P., Neuhauss, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+neurobiology:+how+larval+fish+orient+towards+the+light.">Behavioral neurobiology: how larval fish orient towards the light.</a> Curr. Biol. 20, 159-161 (2010).</li><li>Haug, M. F., Biehlmaier, O., Mueller, K. P., Neuhauss, S. C. <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+in+larval+zebrafish:+behavior+and+histology.">Visual acuity in larval zebrafish: behavior and histology.</a> Front. Zool. 7, 8 (2010).</li><li>Fetcho, J. R., Higashijima, S., McLean, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zebrafish+and+motor+control+over+the+last+decade.">Zebrafish and motor control over the last decade.</a> Brain Res.Rev. 57, 86-93 (2008).</li><li>Connaughton, V. P., Nelson, R., Bender, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+evidence+of+GABAA+and+GABAC+receptors+on+zebrafish+retinal+bipolar+cells.">Electrophysiological evidence of GABAA and GABAC receptors on zebrafish retinal bipolar cells.</a> Vis. Neurosci. 25, 139-153 (2008).</li><li>Kim, Y. J., Nam, R. H., Yoo, Y. M., Lee, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+functional+evidence+of+GABAergic+neurons+in+parts+of+the+brain+of+adult+zebrafish+(Danio+rerio).">Identification and functional evidence of GABAergic neurons in parts of the brain of adult zebrafish (Danio rerio).</a> Neurosci. Lett. 355, 29-32 (2004).</li><li>Sato, Y., Miyasaka, N., Yoshihara, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hierarchical+regulation+of+odorant+receptor+gene+choice+and+subsequent+axonal+projection+of+olfactory+sensory+neurons+in+zebrafish.">Hierarchical regulation of odorant receptor gene choice and subsequent axonal projection of olfactory sensory neurons in zebrafish.</a> J. Neurosci. 27, 1606-1615 (2007).</li><li>Westerfield, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The zebrafish book: a guide for the laboratory use of zebrafish (Brachydanio rerio). , (1993).</li><li>Lazarova, M., Samanin, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potentiation+by+yohimbine+of+pentylenetetrazol-induced+seizures+in+rats:+role+of+alpha+2+adrenergic+receptors.">Potentiation by yohimbine of pentylenetetrazol-induced seizures in rats: role of alpha 2 adrenergic receptors.</a> Pharmacol. Res. Commun. 15, 419-425 (1983).</li><li>Loscher, W., Honack, D., Fassbender, C. P., Nolting, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+technical,+biological+and+pharmacological+factors+in+the+laboratory+evaluation+of+anticonvulsant+drugs.+III.+Pentylenetetrazole+seizure+models.">The role of technical, biological and pharmacological factors in the laboratory evaluation of anticonvulsant drugs. III. Pentylenetetrazole seizure models.</a> Epilepsy res. 8, 171-189 (1991).</li><li>DeMicco, A., Cooper, K. R., Richardson, J. R., White, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+neurotoxicity+of+pyrethroid+insecticides+in+zebrafish+embryos.">Developmental neurotoxicity of pyrethroid insecticides in zebrafish embryos.</a> Toxicol Sci. 113, 177-186 (2010).</li><li>Arnolds, D. 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Denne protokol er blevet anvendt til at måle den neurale aktivitet af voksne zebrafisk in vivo. Med praksis, kan neurale aktivitet observeres konsekvent, selvom karakteristika (amplitude og form af hændelser) af den optagne aktivitet kan variere mellem de enkelte fisk. Udnyttelse af den ekstracellulære optagelse teknik kan forklare denne observation. Fremgangsmåden giver samtidig overvågning af et stort antal celler i en region 1, så variationer i positionen af den primære elektrode kan spille...

I denne artikel er en protokol, der er beskrevet for at opnå in vivo optagelser af neurale aktivitet i voksen zebrafisk. Anvendes ekstracellulære optagelse metoder, som giver spænding målinger af elektrisk aktivitet inden for et lille område af nervevæv. Denne undersøgelsesmetode involverer overvågning af et stort antal celler i en opfører dyr 1. Tidligere har slice optagelser udført i både voksne og larver, som har øje kop præparater og elektroretinogrammet optagelser. Disse eksperimenter er stort set blevet udført til detaljer fysiologiske reaktioner af forskellige sansesystemer 2-5. Indtil for nylig har intakte hjerne præparater kun været til rådighed til udførelse af elektrofysiologi med zebrafisk larve 3,6,7, hvor respiration og oxygendiffusion kan forekomme gennem huden. Vores forberedelse giver den indfødte neurologiske aktivitet en voksen zebrafisk, der skal måles, mens dyret stadig er ved fuld bevidsthed og bevidst of dens omgivelser. Zebrafisk (Danio rerio) aktuelt spiller en fundamental rolle som model for genetiske, toksikologiske, farmakologiske og fysiopatologiske undersøgelse 3. Zebrafisk har fået synlighed inden for neuroscience, fordi de deler omfattende homologi med pattedyr ved de genetiske, neurale og endokrine niveauer 8. Gennem det seneste årti, har standard neuroanatomisk og immunhistokemiske teknikker blevet brugt til at fastlægge det detaljerede karakteristisk organisering af zebrafisk nervesystem 9-12 og af fordelingen af forskellige neurotransmittere 3,8,13. For nylig har forskere flyttet deres fokus til funktionelle studier 14,15, hvoraf mange centrum på adfærdsmæssige processer 16-19 og elektrofysiologiske karakteristika sansesystemer 2,13,20. Et lille antal af disse undersøgelser har koncentreret sig om den elektriske aktivitet i specifikke områder af adult zebrafisk hjerne 21-23, men ikke blev gennemført ved hjælp af en in vivo-tilgang. Denne protokol kan tilpasses til elektrofysiologiske undersøgelser af både spontane og fremkaldte aktivitet inden zebrafisk nervesystem til at beskrive mønstre af aktivitet i bestemte områder af hjernen. Brugen af ​​denne teknik gør det muligt at foretage sammenligninger mellem den neurologiske aktivitet af unge larvestadier og voksne. Endvidere vores protokol tillader sammenligninger mellem genetiske eller farmakologiske ændringer. Sammen med andre metoder, såsom genteknologi eller farmakologiske forsøg, denne metode giver en ny mulighed for funktionel analyse af neuronal kommunikation og plasticitet i den intakte voksent dyr samt for potentielle applikationer, såsom at studere sen debut epilepsi eller neurodegenerative processer.

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		<tr height="43">
			<td height="43" style="height:43px;width:207px;">70% Ethanol</td>
			<td style="width:91px;">Decon Laboratories</td>
			<td style="width:111px;">2750HC</td>
			<td style="width:132px;">Dilute 100% to 70% with DI water</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:207px;">2 M Potassium Chloride</td>
			<td style="width:91px;">J.T. Baker</td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:207px;">2 M Sodium Chloride</td>
			<td style="width:91px;">J.T. Baker</td>
			<td style="width:111px;">3624-05</td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:207px;">0.4% Tris-Buffered Tricaine</td>
			<td style="width:91px;">Sigma-Aldrich</td>
			<td style="width:111px;">E10521</td>
			<td style="width:132px;">pH 7.2-7.4; stored at -20 oC</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:207px;">Pancuronium Bromide</td>
			<td style="width:91px;">Sigma-Aldrich</td>
			<td style="width:111px;">P1918</td>
			<td style="width:132px;">Diluted to 1 &#956;g/&#956;l in 1x phosphate buffered saline</td>
		</tr>
		<tr height="127">
			<td height="127" style="height:127px;width:207px;">Habitat water</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;">pH 7.0-7.4, conductivity of 400-450 &#956;S; maintained by Instant Ocean and Sodium Bicarbonate</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:207px;">Pentylenetetrazol</td>
			<td style="width:91px;">Sigma-Aldrich</td>
			<td style="width:111px;">P6500</td>
			<td style="width:132px;">Diluted to 300 mM in 1x phosphate buffered saline</td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;width:207px;">Nanofil syringe</td>
			<td style="width:91px;">World Precision Instruments, Inc.</td>
			<td style="width:111px;">06A</td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;width:207px;">34 G Beveled needle</td>
			<td style="width:91px;">World Precision Instruments, Inc.</td>
			<td style="width:111px;">NF34BV</td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">Sponge</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;">Small pore and chemical-free</td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:207px;">Foam-backed fine sand paper</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;">5 x 5 cm2 is large enough</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:207px;">9 V Battery</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:207px;">Wires with alligator clips</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;">Need 2</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:207px;">37 cm x 42 cm Kimwipe</td>
			<td style="width:91px;">Kimberly-Clark Professional</td>
			<td style="width:111px;">TW31KEM</td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:207px;">11 cm x 21 cm Kimwipe</td>
			<td style="width:91px;">Kimberly-Clark Professional</td>
			<td style="width:111px;">TW31KWP</td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:207px;">1/8 in diameter tube</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:207px;">1 cm diameter tube</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:207px;">1 mm diameter tube</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">Reducing valve with female Luer lock cap and silicone ferrule</td>
			<td style="width:91px;">Qosina</td>
			<td style="width:111px;">51505</td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">Microscope (Leica MZ APO)</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;">Another microscope can be used</td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;width:207px;">Vanna scissors</td>
			<td style="width:91px;">Roboz Surgical Instruments Co., Inc.</td>
			<td style="width:111px;">15018-10</td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;width:207px;">60 ml Luer lock syringe tubes</td>
			<td style="width:91px;">Becton, Dickinson and Company</td>
			<td style="width:111px;">309653</td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">3-way Stopcocks with Luer connections</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">1-way Stopcock with Luer connection</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">Fisherbrand 100 mm x 15 mm Petri dish</td>
			<td style="width:91px;">Fisher Scientific</td>
			<td style="width:111px;">NC9299146</td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">Fisherbrand 60 mm x 15 mm Petri dish</td>
			<td style="width:91px;">Fisher Scientific</td>
			<td style="width:111px;">S67961</td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:207px;">4 in Borosilicate capillary tube</td>
			<td style="width:91px;">World Precision Instruments</td>
			<td style="width:111px;">TW100F-4</td>
			<td style="width:132px;">Can contain a filament to aid in filling with solution</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:207px;">P-97 Flaming/Brown Micropipette Puller</td>
			<td style="width:91px;">Sutter Instrument Co.</td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">Digidata 1440</td>
			<td style="width:91px;">Molecular Devices</td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">Axon Aloclamp 900A</td>
			<td style="width:91px;">Molecular Devices</td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">Axoclamp software</td>
			<td style="width:91px;">Molecular Devices</td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">HS-9Ax 1U headstage</td>
			<td style="width:91px;">Molecular Devices</td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">0.010 in Silver wire</td>
			<td style="width:91px;">A-M Systems, Inc.</td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">Q-series electrode holder</td>
			<td style="width:91px;">Warner Instruments</td>
			<td style="width:111px;">QSW-A10P</td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:207px;">10 ml Luer lock syringe</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:207px;">1 mm x 15 in Tubing</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;">Connect Luer lock syringe to Q-series electrode holder</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:207px;">Micromanipulator</td>
			<td style="width:91px;">Warner Instruments</td>
			<td style="width:111px;"></td>
			<td style="width:132px;">Need 2</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:207px;">Microsoft-based PC</td>
			<td style="width:91px;">Dell</td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:207px;">Faraday Cage</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:207px;">Air Table</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:207px;">Dissecting Microscope</td>
			<td style="width:91px;"></td>
			<td style="width:111px;"></td>
			<td style="width:132px;"></td>
		</tr>
	</tbody>
</table>

electrophysiological recording in the brain of intact adult zebrafish

Tidligere har elektrofysiologiske studier i voksne zebrafisk været begrænset til at skære præparater eller til øje kop præparater og electrorentinogram optagelser. Dette papir beskriver, hvordan en voksen zebrafisk kan immobiliseres, intuberet og anvendes til in vivo elektrofysiologiske eksperimenter, som muliggør optagelse af neural aktivitet. Immobilisering af den voksne kræver en mekanisme med opløst oxygen til gæller i stedet for buccal og opercular bevægelse. Med vores teknik er dyr immobiliseres og perfunderet med habitat vand for at opfylde dette krav. En craniotomy udføres under tricaine methanesulfonate (MS-222, tricaine) anæstesi til at give adgang til hjernen. Den primære elektrode er derefter anbragt i kraniotomi vinduet til at optage ekstracellulære hjerneaktivitet. Gennem brug af en multitube perfusion system, kan en bred vifte af farmakologiske forbindelser administreres til voksne fisk, og eventuelle ændringer i neural aktivitetkan iagttages. Den metode ikke kun mulighed for observationer, der skal foretages om ændringer i neurologisk aktivitet, men det giver også mulighed for at foretage sammenligninger mellem larve og voksen zebrafisk. Dette giver forskerne mulighed for at identificere de ændringer i neurologisk aktivitet på grund af indførelsen af ​​forskellige forbindelser ved forskellige livsfaser.

Denne protokol er blevet anvendt til at måle den neurale aktivitet af voksne zebrafisk in vivo. Disse elektrofysiologiske optagelser er konsekvent og reproducerbar opnået. Figur 5 viser et repræsentativt eksempel på indfødte og inducerede ændringer af den neurale aktivitet af en voksen zebrafisk når pentylentetrazol (PTZ), en fælles chemoconvulsant 6,7,25,26, er indført i intubation opsætning. Den native neurologisk aktivitet af den voksne zebra...

Watch this Scientific Journal Video about Electrophysiological Recording in the Brain of Intact Adult Zebrafish at JoVE.com

Electrophysiological Recording in the Brain of Intact Adult Zebrafish

Dette papir beskriver, hvordan en voksen zebrafisk kan immobiliseres, intuberet og anvendes til in vivo elektrofysiologiske eksperimenter for at tillade optagelser og manipulation af neural aktivitet i et intakt dyr.

Elektrofysiologiske optagelse i hjernen af ​​intakte Voksen Zebrafisk

Previously, electrophysiological studies in adult zebrafish have been limited to slice preparations or to eye cup preparations and electrorentinogram ...

Research

JoVE Journal

Neuroscience

Electrophysiological Characterization of GFP-Expressing Cell Populations in the Intact Retina

Elektrofysiologiske Karakterisering af GFP-udtrykkende cellepopulationer i den intakte Retina

Studying the physiological properties and synaptic connections of specific neurons in the intact tissue is a challenge for those cells that lack ...

Forebrain Electrophysiological Recording in Larval Zebrafish

Forhjerne Elektrofysiologiske Optagelse i Larvernes zebrafisk

Epilepsy affects nearly 3 million people in the United States and up to 50 million people worldwide. Defined as the occurrence of spontaneous unprovoked ...

Recording Electrical Activity from Identified Neurons in the Intact Brain of Transgenic Fish

Optagelse elektrisk aktivitet fra Identificerede Neuroner i ubeskadiget Brain af transgene fisk

Understanding the cell physiology of neural circuits that regulate complex behaviors is greatly enhanced by using model systems in which this work can be ...

Micromanipulation of Gene Expression in the Adult Zebrafish Brain Using Cerebroventricular Microinjection of Morpholino Oligonucleotides

Mikromanipuleringer af genekspression i Adult zebrafisk Brain Brug Cerebroventricular mikroinjektion af Morpholino oligonukleotider

Manipulation of gene expression in tissues is  required to perform functional studies. In this paper, we demonstrate the  cerebroventricular ...

Simultaneous Electrophysiological Recording and Micro-injections of Inhibitory Agents in the Rodent Brain

Samtidig Elektrofysiologisk optagelse og Micro-injektioner af inhiberende Agents i Rodent Brain

Here we describe a method for the construction of a single-use &#8220;injectrode&#8221; using commercially accessible and affordable parts. A probing ...

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Elektrofysiologiske Metode til Recording Intracellulær Voltage Responses af<em&gt; Drosophila</em&gt; Fotoreceptorer og interneuroner til Light Stimuli<em&gt; In vivo</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

Recording Synaptic Plasticity in Acute Hippocampal Slices Maintained in a Small-volume Recycling-, Perfusion-, and Submersion-type Chamber System

Optagelse synaptisk plasticitet i akut hippocampus skiver vedligeholdes i en lille-volumen genanvendelse - Perfusion- og neddykning-type kammer System

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful ...

Modeling Amyloid-&#946;42 Toxicity and Neurodegeneration in Adult Zebrafish Brain

Modeling Amyloid-ÃƒÆ’Ã…Â½Ãƒâ€šÃ‚Â²42 Toxicity and Neurodegeneration in Adult Zebrafish Brain

Modellering Amyloid-β42 toksicitet og Neurodegeneration i voksen zebrafisk hjerne

Alzheimer's disease (AD) is a debilitating neurodegenerative disease in which accumulation of toxic amyloid-&#946;42 (A&#946;42) peptides leads to ...

Wireless Electrophysiological Recording of Neurons by Movable Tetrodes in Freely Swimming Fish

Trådløs elektrofysiologisk optagelse af neuroner af bevægelige Tetrodes i frit svømmehal

The neural mechanisms governing fish behavior remain mostly unknown, although fish constitute the majority of all vertebrates. The ability to record brain ...

Elektrofysiologiske optagelse i hjernen af ​​intakte Voksen Zebrafisk

Elektrofysiologiske optagelse i hjernen af intakte Voksen Zebrafisk