Name: a large lateral craniotomy procedure for mesoscale wide-field optical imaging of brain activity
Uploaded: 2017-05-07T13:00:00
Description: Watch this Scientific Journal Video about A Large Lateral Craniotomy Procedure for Mesoscale Wide-field Optical Imaging of Brain Activity at JoVE.com

Dette arbejde blev støttet af en naturvidenskab og teknik Forskningsråd of Canada (NSERC) Discovery Grant # 40.352, Campus Alberta for innovation Stol, Alberta Alzheimer Research Program til at MHM, og NSERC CREATE i BIF ph.d. fællesskab og AIHS postgraduate fællesskab til MK. Vi takker Pu Min Wang for udviklingen af ​​denne protokol og til kirurgisk træning, og Behroo Mirza Agha og Di Shao for dyrehold.

<html> Følgende protokol følger Universitet Lethbridge Animal Care udvalg (ACC) retningslinjer, og gennemføres i overensstemmelse med standarderne i den canadiske Rådet om Animal Care (CCAC). 1. Fremstilling <ol><li> For længerevarende studieophold, autoklaveres alle åbne kirurgiske forsyninger og sikre fastholdes, at steriliteten hele operationen. Hvis flere operationer er nødvendige, autoklave mellem operationer. </li><li> Sikre, at der er masser af hjerne-buffer på hånden (minds...

<ol>
	<li>Sigler, A., Mohajerani, M. H., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+rapid+redistribution+of+sensory-evoked+depolarization+through+existing+cortical+pathways+after+targeted+stroke+in+mice.">Imaging rapid redistribution of sensory-evoked depolarization through existing cortical pathways after targeted stroke in mice.</a> Proc Natl Acad Sci U S A. 106 (28), 11759-11764 (2009).</li><li>Shih, A. 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=Two-photon+microscopy+as+a+tool+to+study+blood+flow+and+neurovascular+coupling+in+the+rodent+brain.">Two-photon microscopy as a tool to study blood flow and neurovascular coupling in the rodent brain.</a> J Cereb Blood Flow Metab. 32 (7), 1277-1309 (2012).</li><li>Grinvald, A., Hildesheim, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=VSDI:+a+new+era+in+functional+imaging+of+cortical+dynamics.">VSDI: a new era in functional imaging of cortical dynamics.</a> Nat Rev Neurosci. 5 (11), 874-885 (2004).</li><li>Blinder, P., Shih, A. Y., Rafie, C., Kleinfeld, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topological+basis+for+the+robust+distribution+of+blood+to+rodent+neocortex.">Topological basis for the robust distribution of blood to rodent neocortex.</a> Proc Natl Acad Sci U S A. 107 (28), 12670-12675 (2010).</li><li>Mohajerani, 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=Spontaneous+cortical+activity+alternates+between+motifs+defined+by+regional+axonal+projections.">Spontaneous cortical activity alternates between motifs defined by regional axonal projections.</a> Nat Neurosci. 16 (10), 1426-1435 (2013).</li><li>Mohajerani, M. H., McVea, D. A., Fingas, M., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mirrored+bilateral+slow-wave+cortical+activity+within+local+circuits+revealed+by+fast+bihemispheric+voltage-sensitive+dye+imaging+in+anesthetized+and+awake+mice.">Mirrored bilateral slow-wave cortical activity within local circuits revealed by fast bihemispheric voltage-sensitive dye imaging in anesthetized and awake mice.</a> J Neurosci. 30 (10), 3745-3751 (2010).</li><li>Lippert, M. T., Takagaki, K., Xu, W., Huang, X., Wu, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+voltage-sensitive+dye+imaging+of+rat+cortical+activity+with+high+signal-to-noise+ratio.">Methods for voltage-sensitive dye imaging of rat cortical activity with high signal-to-noise ratio.</a> J Neurophysiol. 98 (1), 502-512 (2007).</li><li>Misgeld, T., Kerschensteiner, M. <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+imaging+of+the+diseased+nervous+system.">In vivo imaging of the diseased nervous system.</a> Nat Rev Neurosci. 7 (6), 449-463 (2006).</li><li>Kerr, J. N., Denk, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+in+vivo:+watching+the+brain+in+action.">Imaging in vivo: watching the brain in action.</a> Nat Rev Neurosci. 9 (3), 195-205 (2008).</li><li>Aronoff, 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=Long-range+connectivity+of+mouse+primary+somatosensory+barrel+cortex.">Long-range connectivity of mouse primary somatosensory barrel cortex.</a> Eur J Neurosci. 31 (12), 2221-2233 (2010).</li><li>McVea, D. A., Mohajerani, M. H., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Voltage-sensitive+dye+imaging+reveals+dynamic+spatiotemporal+properties+of+cortical+activity+after+spontaneous+muscle+twitches+in+the+newborn+rat.">Voltage-sensitive dye imaging reveals dynamic spatiotemporal properties of cortical activity after spontaneous muscle twitches in the newborn rat.</a> J Neurosci. 32 (32), 10982-10994 (2012).</li><li>Sweetnam, 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=Diabetes+impairs+cortical+plasticity+and+functional+recovery+following+ischemic+stroke.">Diabetes impairs cortical plasticity and functional recovery following ischemic stroke.</a> J Neurosci. 32 (15), 5132-5143 (2012).</li><li>Yin, Y. Q., 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+field+recordings+effectively+monitor+the+mouse+cortex+and+hippocampus+under+isoflurane+anesthesia.">In vivo field recordings effectively monitor the mouse cortex and hippocampus under isoflurane anesthesia.</a> Neural Regeneration Research. 11 (12), 1951-1955 (2016).</li><li>Sharp, P. 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=Comparison+of+stimulus-evoked+cerebral+hemodynamics+in+the+awake+mouse+and+under+a+novel+anesthetic+regime.">Comparison of stimulus-evoked cerebral hemodynamics in the awake mouse and under a novel anesthetic regime.</a> Scientific Reports. 5, 12621 (2015).</li><li>Kyweriga, M., Mohajerani, M. H., Kianianmomeni, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optogenetics:+Methods+and+Protocols.">Optogenetics: Methods and Protocols.</a> Methods in Molecular Biology. 1408, 251-265 (2016).</li><li>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=.">.</a> Imaging in neuroscience and development : a laboratory manual. , (2005).</li><li>Vanni, M. P., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesoscale+transcranial+spontaneous+activity+mapping+in+GCaMP3+transgenic+mice+reveals+extensive+reciprocal+connections+between+areas+of+somatomotor+cortex.">Mesoscale transcranial spontaneous activity mapping in GCaMP3 transgenic mice reveals extensive reciprocal connections between areas of somatomotor cortex.</a> J Neurosci. 34 (48), 15931-15946 (2014).</li><li>Xie, 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=Resolution+of+High-Frequency+Mesoscale+Intracortical+Maps+Using+the+Genetically+Encoded+Glutamate+Sensor+iGluSnFR.">Resolution of High-Frequency Mesoscale Intracortical Maps Using the Genetically Encoded Glutamate Sensor iGluSnFR.</a> J Neurosci. 36 (4), 1261-1272 (2016).</li><li>Chan, A. W., Mohajerani, M. H., LeDue, J. M., Wang, Y. T., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesoscale+infraslow+spontaneous+membrane+potential+fluctuations+recapitulate+high-frequency+activity+cortical+motifs.">Mesoscale infraslow spontaneous membrane potential fluctuations recapitulate high-frequency activity cortical motifs.</a> Nat Commun. 6, 7738 (2015).</li><li>Lim, D. 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=In+vivo+Large-Scale+Cortical+Mapping+Using+Channelrhodopsin-2+Stimulation+in+Transgenic+Mice+Reveals+Asymmetric+and+Reciprocal+Relationships+between+Cortical+Areas.">In vivo Large-Scale Cortical Mapping Using Channelrhodopsin-2 Stimulation in Transgenic Mice Reveals Asymmetric and Reciprocal Relationships between Cortical Areas.</a> Front Neural Circuits. 6, (2012).</li><li>Ferezou, 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=Spatiotemporal+dynamics+of+cortical+sensorimotor+integration+in+behaving+mice.">Spatiotemporal dynamics of cortical sensorimotor integration in behaving mice.</a> Neuron. 56 (5), 907-923 (2007).</li><li>Mohajerani, M. H., Aminoltejari, K., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+mini-strokes+produce+changes+in+interhemispheric+sensory+signal+processing+that+are+indicative+of+disinhibition+within+minutes.">Targeted mini-strokes produce changes in interhemispheric sensory signal processing that are indicative of disinhibition within minutes.</a> Proc Natl Acad Sci U S A. 108 (22), E183-E191 (2011).</li><li>Madisen, 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=Transgenic+mice+for+intersectional+targeting+of+neural+sensors+and+effectors+with+high+specificity+and+performance.">Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance.</a> Neuron. 85 (5), 942-958 (2015).</li></ol>

Denne innovative protokol for et stort kranie vindue muliggør samtidig billedbehandling over tidsmæssige og parietale områder af hjernebarken. Kombineret med optisk afbildning, kan det hjælpe at afsløre neurale dynamik inden kortikale områder under spontan og stimulus-induceret aktivitet. Denne ekspansive kraniotomi udsætter også en stor udvidelse af den korticale vaskulatur netværket, herunder den proximale ende af den midterste cerebrale arterie (MCA), muliggør in vivo-billeddannelse af blodstrømme...

Kraniotomi er en standardprocedure, som neurovidenskaberne bruger til at afsløre en del af hjernen. Siden starten af ​​elektrofysiologi har kraniotomi muliggjort usædvanlige gennembrud inden for neurovidenskab. Tæt kortlægning af cerebral cortex med elektroder har ført til forsøg, der tester hypoteser og teorier baseret på disse kort. Vi er for nylig kommet ind i en ny æra, hvor kraniotomi anvendes til in vivo billeddannelse af cortical blodgennemstrømning 1 , 2 , 3 og neurovaskulær arkitektur 4 , der muliggør real-time visualisering af kortikal aktivitet inden for de udsatte områder 5 , 6 , 7 . Selvom mange studier bruger craniotomier kombineret med in vivo optiske billedteknikker til at studere strukturen og funktionen af ​​kortikale neuroner, glia og cortiske vaskulatur 8, 9 er yderligere undersøgelser begrænset af små områder af blotlagt cortex (men se 10). Formålet med denne protokol er at tilvejebringe en fremgangsmåde til at skabe en stor lateral kraniotomi, udsætter hjernebarken fra midterlinjen til squamosal knogle, og strækker sig ud over bregma og lambda. Denne store kraniotomi muliggør samtidig modtagelse af foreningen cortex (retrospenialis, cingulate, og parietale), primær og sekundær motor, somatosensoriske, visuelle og auditive cortex. Denne fremgangsmåde er tidligere blevet kombineret med spænding følsomt farvestof imaging (VSDI) for at undersøge, hvordan flere kortikale områder interagerer med hinanden under spontan og stimulus-induceret cortical aktivitet 5, 11, 12. De mest udfordrende aspekter af denne procedure omfatter placering af hovedetaf dyret, fastsættelse hovedpladen, og undgå blødning mens adskillelse Tindingmusklen fra parietalknoglen. Det må desuden tages i de bore- og kraniet fjernelsesprocesser som skallen kurver i en skrå vinkel.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Heating Pad&#160;</td>
			<td>FHC</td>
			<td>40-90-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine Scissors</td>
			<td>Fine Science Tools</td>
			<td>14058-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps&#160;</td>
			<td>Fine Science Tools</td>
			<td>11251-35</td>
			<td>2 or more pairs are recommended</td>
		</tr>
		<tr>
			<td>Spring scissors</td>
			<td>Fine Science Tools</td>
			<td>15000-00, 15000-10</td>
			<td>1 pair should be designated for dura removal&#160;</td>
		</tr>
		<tr>
			<td>Jet tooth shade powder</td>
			<td>LANG Dental</td>
			<td>Jet Tooth Shade Powder</td>
			<td>to be mixed with the Jet Liquid</td>
		</tr>
		<tr>
			<td>Jet tooth shade liquid</td>
			<td>LANG Dental</td>
			<td>Jet Tooth Shade Liquid</td>
			<td>to be mixed wihth the Jet Powder&#160;</td>
		</tr>
		<tr>
			<td>Drill Heads - Carbide Burs FG 1/4 389</td>
			<td>Midwest Dental</td>
			<td>385201</td>
			<td></td>
		</tr>
		<tr>
			<td>Agarose Powder</td>
			<td>Sigma-Aldrich</td>
			<td>A9793</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelfoam</td>
			<td>Sinclair Dental Canada</td>
			<td>Pfizer Gelfoam</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Western Drug Distribution Centre Ltd</td>
			<td>124125</td>
			<td></td>
		</tr>
		<tr>
			<td>Lidocaine 2% Epinephrine</td>
			<td>Western Drug Distribution Centre Ltd</td>
			<td>125299</td>
			<td></td>
		</tr>
		<tr>
			<td>Dexamethazone 5 mg/mL</td>
			<td>Western Drug Distribution Centre Ltd</td>
			<td>125231</td>
			<td></td>
		</tr>
		<tr>
			<td>Butyl cyanoacrylate glue (VetBond)</td>
			<td>Western Drug Distribution Centre Ltd</td>
			<td>12612</td>
			<td></td>
		</tr>
	</tbody>
</table>

a large lateral craniotomy procedure for mesoscale wide-field optical imaging of brain activity

Kraniotomi er en almindeligvis udføres procedure for at blotlægge hjernen til in vivo eksperimenter. I muse forskning, de fleste laboratorier anvender en lille kraniotomi, typisk 3 mm x 3 mm. Denne protokol indfører en metode til at skabe en væsentlig større 7 mm x 6 mm kraniel vindue udsætter det meste af en cerebral hemisfære over muse tidsmæssige og parietale cortex (f.eks bregma 2,5-4,5 mm, lateral 0 - 6 mm). For at udføre denne operation, skal hovedet vippes ca. 30 ° og meget af Tindingmusklen skal trækkes tilbage. På grund af den store mængde af knoglefjernelse er denne procedure kun beregnet til akutte forsøg med dyret bedøvet under hele operationen og eksperiment. Den største fordel ved denne innovative store laterale kraniel vindue er at give samtidig adgang til både mediale og laterale områder af cortex. Denne store ensidige kraniel vindue kan anvendes til at studere de neurale dynamik mellem celler,samt mellem forskellige kortikale områder ved at kombinere flere elektrodeholdere elektrofysiologiske optagelser, billeddannelse af neuronal aktivitet (fx indre eller ydre billeddannelse), og optogenetic stimulation. Desuden er denne store kraniotomi også udsætter et stort område af kortikale blodkar, der giver mulighed for direkte manipulation af den laterale kortikale vaskulatur.

<html> At undersøge samspillet mellem kortikale områder i et enkelt halvkugle, brugte vi en stor kraniotomi strækker sig tværs over sinus sagittalis og 5 - 6 mm lateralt. Denne kraniel vindue inkluderet primære (motor, somatosensoriske, visuel, auditiv), sekundære (motor, visuelt), og forening (retrospenialis, cingulate, parietal forening) cortex af højre hjernehalvdel (figur 3A). For dette arbejde brugte vi spænding følsom farvestof (VSD) billeddann...

Watch this Scientific Journal Video about A Large Lateral Craniotomy Procedure for Mesoscale Wide-field Optical Imaging of Brain Activity at JoVE.com

A Large Lateral Craniotomy Procedure for Mesoscale Wide-field Optical Imaging of Brain Activity

Denne protokol udgør en metode til at skabe en stor ensidig kraniotomi over tidsmæssige og parietale områder af mus hjernebarken. Dette er specielt nyttigt for sandtidafbildning over et ekspansiv område af en hjernehalvdel.

En stor lateral kraniotomiprocedure for mesoscale bredfelt optisk billeddannelse af hjerneaktivitet

The craniotomy is a commonly performed procedure to expose the brain for in vivo experiments. In mouse research, most labs utilize a small craniotomy, ...

Research

JoVE Journal

Neuroscience

State-Dependency Effects on TMS:  A Look at Motive Phosphene Behavior

Transcranial magnetic stimulation (TMS) is a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate ...

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and ...

Live Imaging of the Ependymal Cilia in the Lateral Ventricles of the Mouse Brain

Live-Imaging af ependymale Cilia i laterale ventrikler af Mouse Brain

Multiciliated ependymal cells line the ventricles in the adult brain. Abnormal function or structure of ependymal cilia is associated with various ...

Modification of a Colliculo-thalamocortical Mouse Brain Slice, Incorporating 3-D printing of Chamber Components and Multi-scale Optical Imaging

Ændring af et Colliculo thalamus Mouse Brain Slice, Omfattende 3D-print af Afdeling Komponenter og Multi-skala Optisk Imaging

The ability of the brain to process sensory information relies on both ascending and descending sets of projections. Until recently, the only way to study ...

Large-scale Three-dimensional Imaging of Cellular Organization in the Mouse Neocortex

Storstilet tredimensional billeddannelse af cellulære organisation i mus Neocortex

The mammalian neocortex is composed of many types of excitatory and inhibitory neurons, each with specific electrophysiological and biochemical ...

Wide-field Single-photon Optical Recording in Brain Slices Using Voltage-sensitive Dye

Wide-felt enkelt-photon optisk optagelse i hjernen skiver ved hjælp af spændings følsom Dye

Wide-field single photon voltage-sensitive dye (VSD) imaging of brain slice preparations is a useful tool to assess the functional connectivity in neural ...

Real-Time fMRI Brain Mapping in Animals

Real-Time fMRI hjernekortlægning hos dyr

Dynamic fMRI responses vary largely according to the physiological conditions of animals either under anesthesia or in awake states. We developed a ...

Activity of Posterior Lateral Line Afferent Neurons during Swimming in Zebrafish

Aktivitet af Posterior Lateral Line Afferent Neuroner under svømning i zebrafisk

Sensory systems gather cues essential for directing behavior, but animals must decipher what information is biologically relevant. Locomotion generates ...

Craniotomy Procedure for Visualizing Neuronal Activities in Hippocampus of Behaving Mice

Craniotomy Procedure for visualisering af neuronale aktiviteter i Hippocampus af barbering mus

Imaging neuronal activities at single-cell resolution in awake behaving animals is a very powerful approach for the investigation of neural circuit ...

In Vivo Wide-Field and Two-Photon Calcium Imaging from a Mouse Using a Large Cranial Window

In Vivo Wide-Field and Two-Photon Calcium Imaging from a Mouse Using a Large Cranial Window

In vivo Bredfelts- og to-foton calciumbilleddannelse fra en mus ved hjælp af et stort kranievindue

Wide-field calcium imaging from the mouse&#39;s neocortex allows one to observe cortex-wide neural activity related to various brain functions. On the ...