Name: recording spatially restricted oscillations in the hippocampus of behaving mice
Uploaded: 2018-07-01T14:30:00
Description: Watch this Scientific Journal Video about Recording Spatially Restricted Oscillations in the Hippocampus of Behaving Mice at JoVE.com

Vi er takknemlige for Karin Winterhalter og Kerstin Semmler for teknisk assistanse. Dette arbeidet ble støttet av klyngen av fortreffelighet BrainLinks - BrainTools (eks 1086) tysk Research Foundation.

Alle metoder som involverer levende dyr har godkjent Regierungspräsidium Freiburg i samsvar med det tyske dyr velferd gjerning.
1. forberedelser
<ol><li>Design og bygge et riktig innsetting verktøy transiently bærer silisium sonden og elektroden koblingen under av implantasjon. Se figur 1 for en eksempel tilpasset bygget innsetting verktøyet.</li>
	<li>Nøye slipp silisium sonde og elektroden kontakten fra emballasjen med keramiske-tipped tang.</li>
	<li>Lø...

<ol>
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Økende bevis viser at hjernen svingninger i hippocampus nevrale kretser oppstår i diskret romlige domener10,11,16. CSD analyse reduserer drastisk påvirkning av volum ledning, en avgjørende forutsetning for studiet av lokale oscillation hendelser. Med denne videoen gir vi en guide til implanting silisium sonder i musen hippocampus for analyse av CSD data. Vi vise eksemplene CSD signalene skarp-bølge krusninger i CA1 og lokal...

Svingninger i LFP er kritisk involvert i informasjonsbehandling av nevrale kretser. De dekker et bredt spekter av frekvenser, alt fra treg bølger (~ 1 Hz) til rask rippel svingninger (~ 200 Hz)1. Forskjellige frekvensbånd forbindes med kognitive funksjoner inkludert minne, emosjonelle behandling og navigasjon2,3,4,5,6,7. Strøm over neuronal membraner utgjør den største delen av LFP signal8. Kasjoner inn cellen (f.eks via aktivering av glutamatergic eksitatoriske synapser) representerer en aktiv gjeldende vask (som kostnad forlater den ekstracellulære mediet). Derimot viser netto flyten av positive ladningen til den ekstracellulære mediet, for eksempel ved aktivering av GABAergic hemmende synapser, en aktiv strømkilde på denne plasseringen. I neuronal dipoler, er gjeldende synker sammen med passiv kilder og omvendt på grunn av kompenserende strøm påvirker membranen kostnad på fjerntliggende områder.
Det elektriske feltet produsert av eksterne nevrale prosesser kan også resultere i betydelig spenning avvisninger er på et opptak-elektrode og kan således anses som feilaktig som et lokalt arrangement. Dette volumet ledning utgjør en alvorlig utfordring til tolkningen av LFP signaler. CSD analyse gir informasjon om lokale gjeldende vasker og kilder underliggende LFP signaler og derfor består av å redusere virkningen av volum ledning8. I laminert strukturer som hippocampus, kan endimensjonal CSD signaler fås ved den andre romlige deriverte av LFP innspilt fra equidistant elektroder arrangert vinkelrett til laminær fly9. Ankomsten av kommersielt tilgjengelig lineær silisium sonder har tillatt forskere å utnytte CSD metoden for studier av lokale oscillation aktivitet prefiks. For eksempel har det vært vist at forskjellige gamma svingninger dukke opp i en lag-spesifikk måte i CA1 området10. Videre har CSD analyse identifisert uavhengig aktiveringspunkt gamma aktivitet i viktigste celle laget av dentate gyrus11. Viktigere, var disse funnene bare i lokale CSD men ikke LFP signaler. CSD analysen gir dermed et kraftig verktøy for å få innsikt i microcircuit driften av hippocampus.
I denne protokollen gir vi en omfattende guide for å få endimensjonal CSD signaler med silisium sonder. Disse metodene kan brukere undersøke lokaliserte oscillation hendelser i hippocampus oppfører mus.

<table border="0" cellpadding="0" cellspacing="0" width="1106">
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		<tr height="42">
			<td height="42" style="height:42px;width:199px;">Crocodile clamp with stand</td>
			<td style="width:381px;">Reichelt Elektronik</td>
			<td style="width:151px;">HALTER ZD-10D</td>
			<td style="width:376px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:199px;">Silicon probe</td>
			<td style="width:381px;">Cambridge Neurotech</td>
			<td style="width:151px;">P-series 32</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Stereoscope</td>
			<td style="width:381px;">Olympus</td>
			<td style="width:151px;">SZ51</td>
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			<td height="84" style="height:84px;width:199px;">Ground screws</td>
			<td style="width:381px;">Screws &#38; More GmbH (screwsandmore.de)</td>
			<td style="width:151px;">DIN 84 A2 M1x2</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Flux</td>
			<td style="width:381px;">Stannol</td>
			<td style="width:151px;">114018</td>
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		<tr height="42">
			<td height="42" style="height:42px;width:199px;">Ceramic-tipped forceps</td>
			<td style="width:381px;">Fine Science Tools</td>
			<td style="width:151px;">11210-60</td>
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			<td height="21" style="height:21px;width:199px;">Paraffine Wax</td>
			<td style="width:381px;">Sigma-Aldrich</td>
			<td style="width:151px;">327204</td>
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			<td height="42" style="height:42px;width:199px;">Cauterizer</td>
			<td style="width:381px;">Fine Science Tools</td>
			<td style="width:151px;">18010-00</td>
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			<td height="21" style="height:21px;width:199px;">Multimeter</td>
			<td style="width:381px;">Uni-T</td>
			<td style="width:151px;">UT61C</td>
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			<td height="21" style="height:21px;width:199px;">Ethanol</td>
			<td style="width:381px;">Carl Roth</td>
			<td style="width:151px;">9065.1</td>
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			<td height="21" style="height:21px;width:199px;">Pasteur pipettes</td>
			<td style="width:381px;">Carl Roth</td>
			<td style="width:151px;">EA65.1</td>
			<td></td>
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			<td height="42" style="height:42px;width:199px;">Heat sterilizer</td>
			<td style="width:381px;">Fine Science Tools</td>
			<td style="width:151px;">18000-45</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Stereotaxic frame</td>
			<td style="width:381px;">David Kopf</td>
			<td style="width:151px;">Model 1900</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Stereotaxic electrode holder</td>
			<td style="width:381px;">David Kopf</td>
			<td style="width:151px;">Model 1900</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Isoflurane</td>
			<td style="width:381px;">Abbvie</td>
			<td style="width:151px;">B506</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Oxygen concentrator</td>
			<td style="width:381px;">Respironix</td>
			<td style="width:151px;">1020007</td>
			<td></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:199px;">Buprenorphine</td>
			<td style="width:381px;">Indivior UK Limited</td>
			<td style="width:151px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Electrical shaver</td>
			<td style="width:381px;">Tondeo</td>
			<td style="width:151px;">Eco-XS</td>
			<td></td>
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			<td height="21" style="height:21px;width:199px;">Heating pad</td>
			<td style="width:381px;">Thermolux</td>
			<td style="width:151px;">463265/-67</td>
			<td></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:199px;">Surgical clamps</td>
			<td style="width:381px;">Fine Science Tools</td>
			<td style="width:151px;">18050-28</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Hydrogen peroxide</td>
			<td style="width:381px;">Sigma-Aldrich</td>
			<td style="width:151px;">H1009</td>
			<td></td>
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			<td height="21" style="height:21px;width:199px;">Sterile cotton wipes</td>
			<td style="width:381px;">Carl Roth</td>
			<td style="width:151px;">EH12.1</td>
			<td></td>
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			<td height="21" style="height:21px;width:199px;">Drill</td>
			<td style="width:381px;">Proxxon</td>
			<td style="width:151px;">Micromot 230/E</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:199px;">21G injection needle</td>
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			<td style="width:151px;">4657527</td>
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			<td height="42" style="height:42px;width:199px;">Phosphate buffer/phosphate buffered saline</td>
			<td style="width:381px;"></td>
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			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Stereotaxic atlas</td>
			<td style="width:381px;">Elsevier</td>
			<td style="width:151px;">9.78012E+12</td>
			<td></td>
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			<td height="42" style="height:42px;width:199px;">Surgical scissors</td>
			<td style="width:381px;">Fine Science Tools</td>
			<td style="width:151px;">14094-11</td>
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			<td height="42" style="height:42px;width:199px;">Surgical forceps</td>
			<td style="width:381px;">Fine Science Tools</td>
			<td style="width:151px;">11272-40</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:199px;">27G injection needles</td>
			<td style="width:381px;">B. Braun</td>
			<td style="width:151px;">4657705</td>
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			<td height="21" style="height:21px;width:199px;">Vaseline</td>
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			<td height="21" style="height:21px;width:199px;">Carprofen</td>
			<td style="width:381px;">Zoetis</td>
			<td style="width:151px;">Rimadyl 50mg/ml</td>
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		<tr height="42">
			<td height="42" style="height:42px;width:199px;">Recording amplifier</td>
			<td style="width:381px;">Intan Technologies</td>
			<td style="width:151px;">C3323</td>
			<td></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:199px;">USB acquisition board</td>
			<td style="width:381px;">Intan Technologies</td>
			<td style="width:151px;">C3004</td>
			<td></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:199px;">Recording cables</td>
			<td style="width:381px;">Intan Technologies</td>
			<td style="width:151px;">C3216</td>
			<td></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:199px;">Electrical commutator</td>
			<td style="width:381px;">Doric lenses</td>
			<td style="width:151px;">HRJ-OE_FC_12_HARW</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Acquisition software</td>
			<td>OpenEphys (www.open-ephys.org)</td>
			<td style="width:151px;">GUI</td>
			<td>allows platform-independent data acquisition</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:199px;">Computer for data acquisition</td>
			<td style="width:381px;"></td>
			<td style="width:151px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Analysis environment</td>
			<td>Python (www.python.org)</td>
			<td style="width:151px;"></td>
			<td>allows platform-independent data analysis</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Urethane</td>
			<td style="width:381px;">Sigma-Aldrich</td>
			<td style="width:151px;"></td>
			<td></td>
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			<td height="21" style="height:21px;width:199px;">Vibratome</td>
			<td style="width:381px;">Leica</td>
			<td style="width:151px;">VT1000</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Microscope slides</td>
			<td style="width:381px;">Carl Roth</td>
			<td style="width:151px;">H868.1</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Cover slips</td>
			<td style="width:381px;">Carl Roth</td>
			<td style="width:151px;">H878.2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Embedding medium</td>
			<td style="width:381px;">Sigma-Aldrich</td>
			<td style="width:151px;">81381-50G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Distilled water</td>
			<td style="width:381px;">Millipore</td>
			<td style="width:151px;">Milli Q</td>
			<td>Table-top machine for the production of distilled water</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:199px;">Tergazyme</td>
			<td style="width:381px;">Alconox</td>
			<td style="width:151px;">Tergazyme</td>
			<td></td>
		</tr>
	</tbody>
</table>

recording spatially restricted oscillations in the hippocampus of behaving mice

Lokale feltet potensial (LFP) kommer fra ion bevegelser over nevrale membraner. Siden spenningen av LFP elektroder gjenspeiler det summerte elektriske feltet av store mengder hjernevev, er trekke ut informasjon om lokale aktivitet utfordrende. Studere neuronal microcircuits, men krever en pålitelig skillet mellom virkelig lokale arrangementer og volum-gjennomførte signaler opprinnelse i fjerne hjernen områder. Gjeldende kilde tetthet (CSD) analyse tilbyr en løsning for dette problemet ved å gi informasjon om gjeldende vasker og kilder i elektrodene. I hjernen områder med laminær cytoarchitecture som hippocampus, kan endimensjonal CSD fås ved å estimere den andre romlige deriverte av LFP. Her beskriver vi en metode for å registrere multilaminar LFPs bruker lineær silisium sonder implantert i dorsal hippocampus. CSD spor beregnes langs personlige shanks av sonden. Denne protokollen dermed beskriver en fremgangsmåte for å løse romlig begrenset nevrale nettverk oscillerende prefiks fritt flytte mus.

Figur 1 illustrerer det innsetting verktøyet for implantering av silisium sonder. Opptak av kronisk implantert silikon sonder målretting CA1 området og granule celle laget av dentate gyrus er vist i figur 2. Vi registrerte LFPs fra sonden marrowbones under fri bevegelse i homecage. For å minimere effekten av volum ledning, ble innhentet signalene konvertert til CSD langs hver skaft av sonden (figur 2B</st...

Watch this Scientific Journal Video about Recording Spatially Restricted Oscillations in the Hippocampus of Behaving Mice at JoVE.com

Recording Spatially Restricted Oscillations in the Hippocampus of Behaving Mice

Denne protokollen beskriver innspillingen av lokale feltet potensialer med flere shank lineær silisium sonder. Konvertering av signaler ved hjelp av gjeldende kilde density analyse kan gjenoppbyggingen av lokale elektrisk aktivitet i musen hippocampus. Med denne teknikken, kan romlig begrenset hjernen svingninger bli studert i fritt flytte mus.

Innspillingen romlig begrenset oscillerende prefiks oppfører seg mus

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

This method can help answer key questions in the neuroscience field such as how neuronal network activities are organized on fine temporal and spatial scales. The main advantage of this technique is that using linear silicon probes current data analysis allows the reconstruction of local oscillations that cannot be easily detected in conventional field potential recordings. To begin this procedure, sterilize the surgical instruments with a hot bead sterilizer, then, wipe all the working surfaces with 70%ethanol.Next, mount the anesthetized animal in a stereotaxic frame by gently inserting ear bars into the ear canal. Once the head of the mouse is stabilized by the ear bars, place a mouthpiece over the snout for continuous isoflurane delivery, then apply ointment to the eyes to prevent drying out. Afterward, place a heating pad underneath the mouse and inject buprenorphine subcutaneously to ensure postoperative analgesia.Subsequently, shave its head and disinfect the skin with 70%ethanol. Using surgical scissors, make an incision on the skin along the midline of the skull and open the skin using surgical clamps. Align the head of the animal with the aid of a stereotaxic alignment tool to level bregma and lambda.There should be less than 50 micrometers of height offset between bregma and lambda. Furthermore, level the head along the medial-lateral axis by measuring the depth at defined distance from bregma on both left and right sides. Adjust the head if it is tilted.Clean the head with 3%hydrogen peroxide and dry it with sterile cotton wipes. Determine the location of the craniotomy relative to bregma using an appropriate stereotaxic atlas. Using a 0.9 millimeter drill head, drill two screw holes in the bone over the cerebellum to place the ground and reference screws.Additionally, drill one to three holes for anchoring screws to stabilize the implant. The location of anchor screws will depend on the location of the craniotomy. Insert the screws in the bone using a suitable screwdriver.Take care not to penetrate into the brain. Now, perform the craniotomy by slowly thinning the skull with the drill in a rectangular area around the implantation side. Frequently moisten the bone with sterilized phosphate buffer.Gently pierce the remaining thinned skull with a 27 gauge injection needle and remove it with a pair of tweezers. Next, carefully pierce the dura mater with a 27 gauge injection needle. Using a pair of tweezers, form a small hook by bending the tip of the needle and use this hook to remove the dura.Then, apply phosphate buffer to prevent the brain surface from drying out. Afterward, mount the electrode insertion tool on a stereotaxic holder and zero the probe on bregma. Move the probe to the stereotaxic coordinates over the craniotomy and slowly penetrate the brain surface.Make sure the probe shafts do not bend and avoid implanting through blood vessels. Then, slowly lower the probe until about 200 micrometers above the desired depth. Next, cover the craniotomy and shanks of the silicon probe with sterilized Vaseline for protection.Subsequently, apply dental cement to fix the base of the probe to the anchoring screw in the skull. Right after cement application, slowly move the probe to the target depth. Advance the last 200 micrometers after applying the cement to reduce lateral movement of the probe and ensure minimal tissue damage in the target area.After the cement has cured, release the probe from the insertion tool by melting the wax with a cauterizer. Then, release the connector board from the insertion device and position it at a suitable place on the skull using a crocodile clamp. Fix the connector board to the skull using dental cement.In the case of probe implantation in the hippocampus, place the connector board on the contralateral parietal bone. Then, solder the ground and reference wires of the connector board to the wires attached to the two screws over the cerebellum. Trim the protective cover to the correct height and place it over the silicon probe.Afterward, fix the cover of the connector board and skull using dental cement, avoiding the skin around the exposed skull. After the surgery, apply appropriate analgesic treatment to the mouse for at least two days. Single-house the mouse to prevent damage to the implant.Recordings from chronically implanted silicon probes targeting the CA1 area are shown here. Current source density analysis is applied along the individual shanks of the probe. In this example, a single sharp wave ripple event leads to a prominent current sink in stratum radiatum of CA1 and in the second example, two distinct sites in the granule cell layer of the dentate gyrus were recorded.The high gamma band was isolated by applying a 60 to 80 hertz band pass filter, revealing local gamma bursts on one of the two recording sites located in the granule cell layer. Note that the local gamma oscillation activity can only be detected after conversion of the recorded signals to CSD. CSD, but not LFP signals, identify the periods of phase and amplitude asynchrony between both recording sites.This segment shows a brief epoch of synchronized gamma activity. The traces here illustrate phase difference and amplitude difference index for LFP and CSD traces. After watching this video, you should have a good understanding of how to implant silicon probes in the hippocampus.

Research

JoVE Journal

Neuroscience

Derivation of Glial Restricted Precursors from E13 mice

Utledning av glial Begrensede Forstadier fra E13 mus

This is a protocol for derivation of glial restricted precursor (GRP) cells from the spinal cord of E13 mouse fetuses. These cells are early precursors ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

Storskala opptak av nevroner med bevegelige Silicon Sonder i oppfører Gnagere

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

Recording Gamma Band Oscillations in Pedunculopontine Nucleus Neurons

Opptak Gamma Band Svingninger i Pedunculopontine Nucleus Nerveceller

Synaptic efferents from the PPN are known to modulate the neuronal activity of several intralaminar thalamic regions (e.g., the ...

Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice

Optogenetic Entrainment av hippocampus Theta svingninger i oppfører seg mus

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

Construction of an Improved Multi-Tetrode Hyperdrive for Large-Scale Neural Recording in Behaving Rats

Byggingen av en forbedret flere sperrende Hyperdrive for store nevrale opptak i oppfører seg rotter

Monitoring the activity patterns of a large population of neurons over many days in awake animals is a valuable technique in the field of systems ...

Long-term Sensory Conflict in Freely Behaving Mice

Langsiktig sensoriske konflikt i fritt oppfører seg mus

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for ...

Investigating Long-term Synaptic Plasticity in Interlamellar Hippocampus CA1 by Electrophysiological Field Recording

Gransker langsiktig Synaptic plastisitet i Interlamellar hippocampus CA1 av elektrofysiologisk Field Recording

The study of synaptic plasticity in the hippocampus has focused on the use of the CA3-CA1 lamellar network. Less attention has been given to the ...

Preparation of Acute Slices from Dorsal Hippocampus for Whole-Cell Recording and Neuronal Reconstruction in the Dentate Gyrus of Adult Mice

Fremstilling av akutte skiver fra Dorsal Hippocampus for helcelleopptak og nevronal rekonstruksjon i Dentate Gyrus av voksne mus

Although the general architecture of the hippocampus is similar along its longitudinal axis, recent studies have revealed prominent differences in ...

Craniotomy Procedure for Visualizing Neuronal Activities in Hippocampus of Behaving Mice

Kraniotomiprosedyre for visualisering av nevronale aktiviteter i Hippocampus av oppførende mus

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