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 Î³ 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 forsøk på mus ble godkjent av Florey Institute dyreetisk komité. 1. Oppsett for Cutting Brain Slices <ol><li> Tilbered en skjære oppløsning bestående av (mM) 125 kolin-Cl, 2,5 KCl, 0,4 CaCl2, 6 MgCl2, 1,25 NaH 2PO 4, 26 NaHCO3, 20 D-glukose mettet med gass karbogen (95% O 2 -5 % CO 2) og en kunstig cerebrospinalvæske (aCSF) opptak oppløsning bestående av (mM) NaCl 125, KCl 2,5, 2 CaCl2, 2 MgCl2,...

<ol>
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En robust metode for å generere CA1 γ svingninger i akutte hjerneskiver er beskrevet. Svingningene generert oppstå fra en lokal krets muliggjør en bedre mulighet for å kontrollere og forstå nevrofysiologiske grunnlaget for nettverks svingninger 12. AMPA-reseptorer, GABAA-reseptorer, I h og T-type Ca2 + kanaler er alt som kreves for γ svingninger i denne modellen. Mens de lokale CA1 svingninger beskrevet her kan bli robust generert dette er avhengig av at de hjerneskiver...

Brain nettverks svingninger forekommer innenfor forskjellige frekvensbånd som korrelerer til atferdstilstander. I gnagere, hippocampus θ svingninger - er (5 10 Hz) observert under utforskende atferd 1,2, mens γ svingninger (20 - 80 Hz) forbinder med ulike kognitive prosesser, herunder persepsjon og oppmerksomhet 3,4. Synkron γ nettverks aktivitet er også implisert i patologien av forstyrrelser slik som epilepsi og schizofreni 5,6. For eksempel er γ svingninger antas å tilsvare områder av kortikale epileptiske foci 5,7,8 og kan brukes som markører for pharmacosensitivity eller motstand, to viktige områder for undersøkelse i epilepsi forskning 9. Hippocampus hjernen skive er en modell som har vært mye brukt til å undersøke nettverksaktivitet 10-12. Ulike protokoller har blitt utviklet for å generere γ svingninger i hjerneskiver som vanligvis jegNvolve farmakologisk modulering slik som lav Mg2 +, 4-aminopyridin (4AP) bicuculline, og kainsyre 12-17. Mangler av farmakologisk utløste svingninger er at de opptrer tilfeldig etter medikament påføring og er ikke pålitelig genereres eller stabil over tid. Elektrisk utløst γ svingninger overkomme mange av disse problemer og har også fordelen av å være tidsmessig låst til den stimulerende arrangementet slik at for episodisk registrering og analyse. Her en protokoll er beskrevet for å generere CA1 γ svingninger ved å levere en tetanic stimulering til stratum Oriens i hippocampus skive.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>4-(N-Ethyl-N-phenylamino)-1,2- dimethyl-6-(methylamino) pyrimidinium chloride (ZD7288)</td>
			<td>Sigma-Aldrich</td>
			<td>Z3777</td>
			<td></td>
		</tr>
		<tr>
			<td>Biuculline</td>
			<td>Sigma-Aldrich</td>
			<td>14340</td>
			<td></td>
		</tr>
		<tr>
			<td>6-cyano-7-nitroquinoxa- line-2,3-dione (CNQX)</td>
			<td>Sigma-Aldrich</td>
			<td>C127</td>
			<td></td>
		</tr>
		<tr>
			<td>Nickel</td>
			<td>Sigma-Aldrich</td>
			<td>266965</td>
			<td></td>
		</tr>
		<tr>
			<td>Carbamazepine</td>
			<td>Sigma-Aldrich</td>
			<td>C4024</td>
			<td></td>
		</tr>
		<tr>
			<td>(2R)-amino-5-phosphonopentano-ate (APV)</td>
			<td>Tocris Bioscience</td>
			<td>0105</td>
			<td></td>
		</tr>
		<tr>
			<td>Retigabine</td>
			<td>ChemPacific</td>
			<td>150812-12-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Choline-Cl</td>
			<td>Sigma Aldrich</td>
			<td>C1879-5KG</td>
			<td></td>
		</tr>
		<tr>
			<td>KCl</td>
			<td>Sigma Aldrich</td>
			<td>P9333-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>NaH2PO4</td>
			<td>Sigma Aldrich</td>
			<td>S9638-250G</td>
			<td></td>
		</tr>
		<tr>
			<td>NaHCO3</td>
			<td>Sigma Aldrich</td>
			<td>S6297-250G</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Sigma Aldrich</td>
			<td>S7653-5KG</td>
			<td></td>
		</tr>
		<tr>
			<td>Glucose</td>
			<td>Sigma Aldrich</td>
			<td>G8270-1KG</td>
			<td></td>
		</tr>
		<tr>
			<td>CaCl2.2H2O</td>
			<td>Sigma Aldrich</td>
			<td>223506-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>MgCl2.6H2O</td>
			<td>Sigma Aldrich</td>
			<td>M2670-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrode glass</td>
			<td>Harvard Apparatus&#160;</td>
			<td>GC150F-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Concentric bipolar stimulating metal electrode&#160;</td>
			<td>FHC</td>
			<td>CBBPF75</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital Isolator</td>
			<td>Getting Instruments</td>
			<td>Model BJN8-9V1&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Model 1800 amplifier</td>
			<td>A-M systems</td>
			<td>Model 1800 amplifier</td>
			<td></td>
		</tr>
		<tr>
			<td>Digitizer</td>
			<td>National Intruments</td>
			<td>NI USB-6211</td>
			<td></td>
		</tr>
		<tr>
			<td>Vibrotome</td>
			<td>Leica</td>
			<td>VT1200s</td>
			<td></td>
		</tr>
	</tbody>
</table>

generation of local ca1 &#947; oscillations by tetanic stimulation

Nevrale nettverk svingninger er viktige funksjoner i hjernens aktivitet i helse og sykdom, og kan moduleres av en rekke klinisk brukt narkotika. En protokoll er anordnet for å generere en modell for å studere CA1 γ oscillasjoner (20 - 80 Hz). Disse γ svingninger er stabile i minst 30 minutter og avhenger av stimulerende og hemmende synaptisk aktivitet i tillegg til aktivering av pacemaker strømninger. Tetanically stimulert oscillasjoner har en rekke av reproduserbar og lett målbare karakteristikker inkludert pigg teller, oscillation varighet, ventetid og frekvens som rapporterer på nettverkstilstand. Fordelene med de elektrisk stimulerte svingninger inkluderer stabilitet, reproduserbarhet og episodisk oppkjøpet gjør det mulig robust karakterisering av nettverksfunksjonen. Denne modellen av CA1 γ svingninger kan brukes til å studere cellulære mekanismer og systematisk undersøke hvordan nettverks neuronal aktivitet er endret på sykdom og av medikamenter.Sykdomstilstand farmakologi lett kan innlemmes ved bruk av hjerneskiver fra genmodifiserte eller intervensjonsdyremodeller for å aktivere en rekke medikamenter som spesifikt retter sykdomsmekanismer.

Tetanic stimulering av stratum Oriens generert robuste og reproduserbare γ svingninger (35,4 ± 2,2 Hz), se figur 3B. For å demonstrere at oscillasjonene ble samlet inn i det lokale nettverket CA1 inngangene fra CA3 ble kuttet ved å skjære stykket i CA2 region ved hjelp av en bøyd 32 G nål. Oscillasjonen eiendommer i de skjære skiver seg ikke fra de uslipte skiver (p = 0,85; skjære skiver 6,16 ± 1,1 pigger, n = 6; uncut skiver 5,89 ± 0,8 pigger, n = 6), noe som indikerer at svingninger generer...

Watch this Scientific Journal Video about Generation of Local CA1 Î³ Oscillations by Tetanic Stimulation at JoVE.com

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

Svingninger er grunnleggende nettverksegenskaper og er modulert av sykdom og medikamenter. Studerer hjerne-slice svingninger tillater karakterisering av isolerte nettverk under kontrollerte forhold. Protokoller for utarbeidelse av akutte hjerneskiver for fremkaller CA1 γ svingninger.

Generasjon Lokale CA1 γ Svingninger av tetanic Stimulering

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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

Disseksjon av lokale Ca2 + signaler i kulturperler celler av membran målrettede Ca2 + indikatorer

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