Name: generation of local ca1 &#947; oscillations by tetanic stimulation
Uploaded: 2015-08-14T13:00:00
Description: Watch this Scientific Journal Video about Generation of Local CA1 ÃŽÂ³ Oscillations by Tetanic Stimulation at JoVE.com

Supported by APA to RJH, NHMRC program grant 400121 to SP, and NMHRC fellowship 1005050 to SP. CAR acknowledges the support of the ARC (FT0990628) and the DOWD fellowship scheme. The Florey Institute of Neuroscience and Mental Health is supported by Victorian State Government infrastructure funds.

Alle forsøg på mus blev godkendt af Florey Institute dyreetik komité. 1. Opsætning til Skæring Brain Skiver <ol><li> Der fremstilles en skærende opløsning omfattende (mM) 125 Cholin-CI, 2,5 KCI, 0,4 CaCl2, 6 MgCl2, 1,25 NaH 2 PO 4, 26 NaHCO3, 20 D-glucose mættet med carbogen gas (95% O2 -5 % CO 2) og en kunstig cerebrospinalvæske (aCSF) optagelse opløsning bestående af (mM) 125 NaCl, 2,5 KCI, 2 CaC...

<ol>
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En robust metode til at generere CA1 γ svingninger i akutte hjernen skiver er beskrevet. Svingninger genereret udspringer fra en lokal kreds muliggør en bedre mulighed for at styre og forstå neurofysiologiske grundlag af net svingninger 12. AMPA-receptorer, GABAA-receptorer, I h og T-typen Ca2 + kanaler er alle nødvendige for γ svingninger i denne model. Mens de lokale CA1 oscillationer beskrevet her kan robust genereret afhænger sikre, at hjernen skiver er sunde. Et afg...

Brain netværk svingninger forekomme inden distinkte frekvensbånd, der korrelerer til adfærdsmæssige tilstande. Hos gnavere hippocampus θ svingninger - er (5 10 Hz) observeret under sonderende adfærd 1,2, mens γ svingninger (20-80 Hz) associeret med forskellige kognitive processer, herunder perception og opmærksomhed 3,4. Synkrone γ netværksaktivitet er også impliceret i patologien af lidelser såsom epilepsi og skizofreni 5,6. For eksempel er γ svingninger menes at svare til områder af kortikale epileptiske foci 5,7,8 og kan anvendes som markører for pharmacosensitivity eller resistens, to vigtige områder af undersøgelsen i epilepsi forskning 9. Den hippocampale hjerne skive er en model, der har været almindeligt anvendt til at undersøge netværksaktivitet 10-12. Forskellige protokoller er blevet udviklet til at generere γ svingninger i hjernen skiver, der typisk involve farmakologisk modulation såsom lav Mg2 +, 4-aminopyridin (4AP), bicucullin og kaininsyre 12-17. Mangler af farmakologisk udløste svingninger er, at de opstår tilfældigt efter narkotika ansøgning og er ikke pålideligt genereres eller stabile over tid. Elektrisk udløsning γ oscillationer overvinde mange af disse problemer og har også den fordel, at tidsmæssigt låst til stimulerende begivenhed giver mulighed for episodisk registrering og analyse. Her en protokol er beskrevet til frembringelse af CA1 γ svingninger ved at levere en tetanic stimulation til stratum Oriens i hippocampale udsnit.

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			<td>Sigma-Aldrich</td>
			<td>Z3777</td>
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			<td>Sigma-Aldrich</td>
			<td>14340</td>
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			<td>6-cyano-7-nitroquinoxa- line-2,3-dione (CNQX)</td>
			<td>Sigma-Aldrich</td>
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			<td>Nickel</td>
			<td>Sigma-Aldrich</td>
			<td>266965</td>
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			<td>Carbamazepine</td>
			<td>Sigma-Aldrich</td>
			<td>C4024</td>
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			<td>(2R)-amino-5-phosphonopentano-ate (APV)</td>
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			<td>Choline-Cl</td>
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			<td>KCl</td>
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			<td>P9333-500G</td>
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			<td>NaH2PO4</td>
			<td>Sigma Aldrich</td>
			<td>S9638-250G</td>
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			<td>NaHCO3</td>
			<td>Sigma Aldrich</td>
			<td>S6297-250G</td>
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			<td>NaCl</td>
			<td>Sigma Aldrich</td>
			<td>S7653-5KG</td>
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			<td>Glucose</td>
			<td>Sigma Aldrich</td>
			<td>G8270-1KG</td>
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			<td>CaCl2.2H2O</td>
			<td>Sigma Aldrich</td>
			<td>223506-500G</td>
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			<td>MgCl2.6H2O</td>
			<td>Sigma Aldrich</td>
			<td>M2670-500G</td>
			<td></td>
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			<td>Electrode glass</td>
			<td>Harvard Apparatus&#160;</td>
			<td>GC150F-10</td>
			<td></td>
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			<td>Concentric bipolar stimulating metal electrode&#160;</td>
			<td>FHC</td>
			<td>CBBPF75</td>
			<td></td>
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			<td>Digital Isolator</td>
			<td>Getting Instruments</td>
			<td>Model BJN8-9V1&#160;</td>
			<td></td>
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			<td>Model 1800 amplifier</td>
			<td>A-M systems</td>
			<td>Model 1800 amplifier</td>
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			<td>Digitizer</td>
			<td>National Intruments</td>
			<td>NI USB-6211</td>
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			<td>Vibrotome</td>
			<td>Leica</td>
			<td>VT1200s</td>
			<td></td>
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generation of local ca1 &#947; oscillations by tetanic stimulation

Neuronale netværk svingninger er vigtige elementer i hjernens aktivitet i sundhed og sygdom, og kan moduleres af en række klinisk anvendte lægemidler. En protokol er tilvejebragt til at generere en model til undersøgelse af CA1 γ svingninger (20-80 Hz). Disse γ svingninger er stabile i mindst 30 minutter og afhænger excitatorisk og inhibitorisk synaptisk aktivitet udover aktivering af pacemaker strømme. Tetanically stimulerede svingninger har en række reproducerbare og let kvantificerbare karakteristika, herunder spike tæller, oscillation varighed, ventetid og frekvens, der rapporterer på netværket tilstand. Fordelene ved de elektrisk stimulerede svingninger omfatter stabilitet, reproducerbarhed og episodisk erhvervelse muliggør robust karakterisering af netværk funktion. Denne model af CA1 γ svingninger kan anvendes til at studere cellulære mekanismer og systematisk undersøge, hvordan neuronal netværksaktivitet er ændret i sygdom og af medikamenter.Sygdomstilstand farmakologi kan let inkorporeres ved anvendelse af hjerneskiver fra genetisk modificerede eller interventionelle dyremodeller for at muliggøre udvælgelse af lægemidler, som specifikt retter sygdomsmekanismer.

Tetanic stimulering af stratum Oriens genereret robuste og reproducerbare γ svingninger (35,4 ± 2,2 Hz), se figur 3B. For at demonstrere, at de svingninger blev genereret inden for den lokale CA1 netværk input fra CA3 blev afbrudt ved at skære skive i CA2 regionen ved hjælp af en bøjet 32 ​​G nål. Svingningerne egenskaber i de afskårne skiver adskilte sig ikke fra de uslebne skiver (p = 0,85; cut skiver 6.16 ± 1,1 spikes, n = 6; uslebne skiver 5,89 ± 0,8 spikes, n 6 =), hvilket indikerer, ...

Watch this Scientific Journal Video about Generation of Local CA1 ÃŽÂ³ Oscillations by Tetanic Stimulation at JoVE.com

Generation of Local CA1 ÃƒÅ½Ã‚Â³ Oscillations by Tetanic Stimulation

Svingninger er grundlæggende netværksegenskaber og moduleres af sygdom og narkotika. At studere hjerne-slice svingninger tillader karakterisering af isolerede net under kontrollerede forhold. Protokoller om tilberedning af akutte hjernen skiver for at fremkalde CA1 γ svingninger.

Generation af Lokale CA1 γ Svingninger ved tetanisk Stimulation

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Generation of Local CA1 &#947; Oscillations by Tetanic Stimulation

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Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice

Optogenetic medrivning af hippocampus Theta svingninger i opfører sig mus

Extensive data on relationships of neural network oscillations to behavior and organization of neuronal discharge across brain regions call for new tools ...

Recording Spatially Restricted Oscillations in the Hippocampus of Behaving Mice

Indspilning rumligt begrænset svingninger i Hippocampus adfærd mus

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed ...

Dissection of Local Ca2+ Signals in Cultured Cells by Membrane-targeted Ca2+ Indicators

Dissection of Local Ca2+ Signals in Cultured Cells by Membrane-targeted Ca2+ Indicators

Dissektion af lokale Ca2 + signaler i kulturperler celler af membran-målrettet Ca2 + indikatorer

Calcium ion (Ca2+) is a universal intracellular messenger molecule that drives multiple signaling pathways, leading to diverse biological outputs. The ...