Name: examining monosynaptic connections in drosophila using tetrodotoxin resistant sodium channels
Uploaded: 2018-02-14T14:00:00
Description: Watch this Scientific Journal Video about Examining Monosynaptic Connections in Drosophila Using Tetrodotoxin Resistant Sodium Channels at JoVE.com

Vi vil gjerne takke Joshua Singer, Jonathan Schenk, samt gjennomtenkt korrekturlesere på manuskriptet. Vi vil også gjerne takke Ben White og Harold Zakon for diskusjoner om teknikken. Jonathan Schenk gitt dataene for figur 3A. Dette arbeidet ble støttet av en Whitehall stiftelse stipend og en NIH R21 til QG.

1. klargjør fluer og identifisere GAL4 promoter linjer
<ol><li>Velg en GAL4 promoter linje, og krysse den med fluer bærer både transgene UAS-NaChBac og en optogenetic transgene for aktivisering. Merk: UAS-CsChrimson11 er valgt på grunn av sin høye konduktans og enkel som aktiverer NaChBac-mediert handling potensialer. Både UAS-NaChBac og UAS-CsChrimson er tilgjengelig fra Bloomington aksjer depotet.</li>
	<li>Forberede all-trans retinal som lager løsning i etanol (35 mM) og lagre løsn...

<ol>
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K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole+Cell+Recordings+from+Brain+of+Adult+Drosophila.">Whole Cell Recordings from Brain of Adult Drosophila.</a> J Vis Exp. , (2007).</li><li>Wilson, R. I., Turner, G. C., Laurent, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transformation+of+olfactory+representations+in+the+Drosophila+antennal+lobe.">Transformation of olfactory representations in the Drosophila antennal lobe.</a> Science. 303, 366-370 (2004).</li><li>Kazama, H., Wilson, R. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Homeostatic+Matching+and+Nonlinear+Amplification+at+Identified+Central+Synapses.">Homeostatic Matching and Nonlinear Amplification at Identified Central Synapses.</a> Neuron. 58, 401-413 (2008).</li><li>Becnel, J., 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=DREADDs+in+Drosophila:+a+pharmacogenetic+approach+for+controlling+behavior,+neuronal+signaling,+and+physiology+in+the+fly.">DREADDs in Drosophila: a pharmacogenetic approach for controlling behavior, neuronal signaling, and physiology in the fly.</a> Cell Reports. 4, 1049-1059 (2013).</li><li>Armbruster, B. N., Li, X., Pausch, M. H., Herlitze, S., Roth, B. 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B., 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=Monosynaptic+Glutamatergic+Activation+of+Locus+Coeruleus+and+Other+Lower+Brainstem+Noradrenergic+Neurons+by+the+C1+Cells+in+Mice.">Monosynaptic Glutamatergic Activation of Locus Coeruleus and Other Lower Brainstem Noradrenergic Neurons by the C1 Cells in Mice.</a> J Neurosci. 33, 18792-18805 (2013).</li><li>Petreanu, L., Mao, T., Sternson, S. M., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+subcellular+organization+of+neocortical+excitatory+connections.">The subcellular organization of neocortical excitatory connections.</a> Nature. 457, 1142-1145 (2009).</li><li>Kloppenburg, P., Ferns, D., Mercer, A. 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TERPS analyse komplimenter gjeldende teknikker for krets sprengning ved å aktivere gjenkjenning av synapser mellom identifiserte neurons. Spesielt avslører tilnærming monosynaptic tilkoblinger forstand stanse handling potensialer med TTX under gjenoppretting excitability i Velg innbyggere neurons med TTX-ufølsom natrium kanalen NaChBac. Synaptiske utgivelsen er brakt frem av optogenetic stimulering mens postsynaptic hendelser overvåkes med hele celle innspillinger. TERPS skiller seg fra andre tilnærminger som rekke...

Et hovedmål for nevrovitenskap er å tilordne forbindelsene mellom neurons å forstå hvordan informasjon flyter gjennom kretser. Mange tilnærminger og ressurser har blitt tilgjengelig til å teste funksjonelle tilkoblingen mellom nerveceller i en rekke modell systemer1,2. For å generere de mest nøyaktige koblingsskjema bruker elektrofysiologi, er det viktig å løse om observerte forbindelser mellom to celler som er direkte og monosynaptic indirekte og polysynaptic. En gull-standard for å gjøre dette skillet i pattedyr neurons er å måle ventetiden mellom én action potentials i de presynaptic cellene og utbruddet av eksitatoriske postsynaptic strømmer (EPSCs) i andre Nevron. Monosynaptic tilkoblinger bør ha kort ventetid av noen millisekunder og lav variasjon3. Denne tilnærmingen kan være komplisert i virvelløse neurons fordi synapser kan oppstå i sin dendrites langt fra somatiske innspillingen området, forårsaker forsinkelser i påvisning på grunn av lange electrotonic avstander mellom postsynaptic conductances og opptak elektroden. Slike forsinkelser kan introdusere tvetydighet om poly-kontra monosynaptic bidrag. I tillegg liten synaptic hendelser kan forfalle før området somatiske innspillinger og kjøring sterkere presynaptic aktivitet, er sannsynlig å rekruttere polysynaptic hendelser.
Ulike teknikker er utviklet for å teste for monosynaptic forbindelser i dyr. En tilnærming bruker høy divalent kasjon løsninger (Hi-Di) som inneholder overflødig Mg++ og Ca++. Denne løsningen blokkerer polysynaptic tilkoblinger ved å redusere utgivelsen sannsynlighet og økende handling potensial terskel å favorisere gjenkjenning av monosynaptic inn4,5. Bestemme nøyaktig forholdet mellom Mg++ til Ca++ kreves, men er ikke trivielt og polysynaptic bidrag kan vare i selv beskjeden stimulering6. En alternativ tilnærming kalt GFP rekonstituering over Synaptic partnere (tak) drar nytte av nærhet mellom før og postsynaptic membraner på synapser å antyde en monosynaptic forbindelse 7. Her en komponent av grønne fluorescerende protein (GFP) er uttrykt i en Nevron, og komplementære fragmentet av molekylet er uttrykt i en antatte postsynaptic partner. Tilstedeværelsen av fluorescens angir at to neurons i nok nærheten tillater rekonstituering av GFP molekyl og innebærer eksistensen av en synapse. FORSTÅELSE kan rapportere synapser feilaktig, skjønt, hvis to celler har tett apposed membraner, som i en nerve eller fascicle. Varianter av forståelse eliminere slike falske positiver ved tethering presynaptic fragmentet av GFP direkte til synaptobrevin, slik at rekonstituering bare på aktive synapser8. Mens forståelse og variantene har vært medvirkende i å bestemme funksjonelle tilkobling i Drosophila CNS, kan noen tilkoblinger ikke gjøres synlig av forståelse om avstandene mellom pre og postsynaptic partnere er relativt stor. Dette er spesielt relevant i vurderingen av volum overføring forbundet med neuromodulation9 eller GABAergic hemming10.
Her er en komplementær og romanen teknikken demonstrert for testing direkte monosynaptic forbindelser i Drosophila CNS. Denne metoden kalles Tetrodotoxin foretatt motstand for undersøkelser synapser (TERPS), fungerer ved co uttrykk for tetrodotoxin (TTX)-resistente natrium kanal, NaChBac og optogenetic aktivator i en presynaptic celle under opptak fra antatte postsynaptic partnere9. I nærvær av TTX undertrykkes alle action potensial-mediert aktivitet i cellene ikke uttrykke NaChBac. NaChBac kanalen gjenoppretter selektivt excitability i målrettede presynaptic cellen(e), tillater lys-utløste synaptic overføring. Denne metoden tillater sterk aktivering av presynaptic neurons, slik at tilkoblinger kan løses postsynaptically ved somatiske opptak samtidig redusere sannsynligheten for rekruttering polysynaptic kretser. Viktigere, denne teknikken tillater studiet av volum overføring og avslører spredning av senderen fra en enkelt Nevron i en krets. I tillegg kan TERPS avsløre av kjemisk natur tilkoblingen gjennom konvensjonelle farmakologi. TERPS er egnet for bruk i noen modellsystem som tillater transgene uttrykket TTX tittelen natrium kanaler.

<table border="0" cellpadding="0" cellspacing="0" width="946">
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">UAS-csChrimson</td>
			<td style="width:219px;">Bloomington Drosophila Stock Center</td>
			<td style="width:344px;">55135</td>
			<td style="width:167px;">Used as a neural activator</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">UAS-NaChBac</td>
			<td>Bloomington Drosophila Stock Center</td>
			<td>9466</td>
			<td style="width:167px;">Resotores excitibility in cells in TTX</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Tetrodotoxin</td>
			<td>Tochris</td>
			<td>1078</td>
			<td style="width:167px;">Special permission may be needed to purchase TTX as it is a controlled substance</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">all trans-Retinal</td>
			<td>Sigma-Aldrichall trans-Retinal</td>
			<td>R2500</td>
			<td style="width:167px;">Require co-factor for channelrhodopsin</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Weldable 321 Stainless Steel&#160;Sheet,&#160;0.002&#34;&#160;Thick,&#160;10&#34;&#160;Wide</td>
			<td>McMaster Carr</td>
			<td>3254K7</td>
			<td style="width:167px;">Used to make custom fly holder. Custom foil can be laser cut at pololu.com from the provided PDF file</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Dissection Microscope</td>
			<td>Zeiss</td>
			<td>Stemi 2000-C</td>
			<td style="width:167px;">Used for dissection of preparation</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Waxer</td>
			<td>Almore</td>
			<td>Eectra Waxer 66000</td>
			<td style="width:167px;">Used during dissection to secure fly in foil</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Paraffin Wax</td>
			<td>Joann</td>
			<td>4917217</td>
			<td style="width:167px;">Used with waxer</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Number 5 forceps</td>
			<td>Fine Science Tools</td>
			<td>#5CO</td>
			<td style="width:167px;">Used for dissection and desheathing</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Dissection Scissors</td>
			<td>Fine Science Tools</td>
			<td>15001-08</td>
			<td style="width:167px;">Used to remove parts of the cuticle during dissection</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Tungsten wire</td>
			<td>A-M Systems</td>
			<td>797500</td>
			<td style="width:167px;">Use with electrolysis to make sharpened needles for dissection</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Reciculating Peristaltic Pump</td>
			<td>Simply Pumps (Amazon)</td>
			<td>PM200S</td>
			<td style="width:167px;">for recirculating TTX</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Speed Controller for peristaltic pump</td>
			<td>Zitrades (Amazon)</td>
			<td style="width:344px;">N/A</td>
			<td style="width:167px;">PWM Dimming Controller For LED Lights or Ribbon, 12 Volt 8 Amp,Adjustable Brightness Light Switch Dimmer Controller DC12V 8A 96W for Led Strip Light B</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Versa-Mount&#160;Precision Compressed Air Regulator</td>
			<td>McMaster Carr</td>
			<td style="width:344px;">1804T1</td>
			<td style="width:167px;">For applying positive pressure during patching</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Glass capilaries</td>
			<td>World Precision Instruments</td>
			<td style="width:344px;">TW150F-3</td>
			<td style="width:167px;">For patch pipettes</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Multipurpose Gauge</td>
			<td>McMaster Carr</td>
			<td>3846K431</td>
			<td style="width:167px;">Gauge for pressure regulator</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Electrophysiology Camera</td>
			<td>Dage MTI&#160;</td>
			<td>IR-1000</td>
			<td style="width:167px;">Any camera that works in the IR range (850 nm) will work. You do not want to use red illumination as this can activate csChrimson</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">IR LED</td>
			<td>Thorlabs&#160;</td>
			<td>M850F2</td>
			<td style="width:167px;">For oblique illumination</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">fiber optic for IR LED</td>
			<td>Thorlabs</td>
			<td>M89L01</td>
			<td style="width:167px;">Couples to IR LED</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Objective lens&#160;</td>
			<td>Olympus 40X</td>
			<td>LUMPlanFLN</td>
			<td style="width:167px;">This can be used on most microscipes and works well for visualizing fly neurons.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Amber LED</td>
			<td>Thorlabs</td>
			<td>M590L3d</td>
			<td style="width:167px;">For visualizing RFP and mCherry</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Blue LED</td>
			<td>Thorlabs</td>
			<td>M470L3d</td>
			<td style="width:167px;">For visualizing GFP</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">GFP filter set</td>
			<td>Chroma</td>
			<td>49011</td>
			<td style="width:167px;">For visualizing GFP or stimulating channel2rhodopsin</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Custom mCherry Filter Set</td>
			<td>Chroma</td>
			<td style="width:344px;">&#160;et580/25x and t600lpxr (from the 49306 set) but with an et610lp barrier/emission optic</td>
			<td style="width:167px;">Use only if you wish to patch identified neurons with channel2rhodopsin</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Dichroic to combine Amber and blue LED</td>
			<td>Thorlabs</td>
			<td style="width:344px;">DMLP550R</td>
			<td style="width:167px;">Use only patch under mCherry and excite channel2rhodopsin with blue light.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Red LED</td>
			<td>LEDSupply</td>
			<td>Cree XPE 620 - 630 nm</td>
			<td style="width:167px;">Used to drive csChrimson</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">LED Driver</td>
			<td>LEDSupply</td>
			<td>3021-D-E-1000</td>
			<td style="width:167px;">Used to drive LEDs for optogenetic stimulation</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Manipulator</td>
			<td>Sutter Instruments</td>
			<td>MP-225</td>
			<td style="width:167px;">Used to position pipette during recordings</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Patchclamp Amplifier</td>
			<td>A-M Systems</td>
			<td>Model 2400</td>
			<td style="width:167px;">An equivalent amplifier is suitable</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Bessel Filter</td>
			<td>Warner Instruments&#160;</td>
			<td>LPF 202A</td>
			<td style="width:167px;">Auxillary filter used to filter current trace to oscilloscope during patching.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Data Acquisition System&#160;</td>
			<td>National Instruments</td>
			<td>NI PCIe-6321&#160;&#160;&#160;&#160;&#160;&#160; 781044-01</td>
			<td style="width:167px;">Used to record data from amplifier to computer</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Connector Block - BNC Terminal BNC-2090A</td>
			<td>National Instruments</td>
			<td>779556-01</td>
			<td style="width:167px;">Used to connect amplifier to DAQ card.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Steel Foil&#160;</td>
			<td>McMaster Carr</td>
			<td>3254K7</td>
			<td style="width:167px;">Steel foil for custom recording chamber</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Magnets</td>
			<td>K&#38;J Magnetics</td>
			<td>D42</td>
			<td style="width:167px;">To secure recording chamber to ring stand</td>
		</tr>
		<tr height="100">
			<td height="100" style="height:100px;">1/16&#34; Cell Cast Acrylic Clear</td>
			<td>Pololu</td>
			<td></td>
			<td style="width:167px;">Used to make custom recording chamber. Acrylic can be laser cut at pololu.com from the provided PDF file</td>
		</tr>
	</tbody>
</table>

examining monosynaptic connections in drosophila using tetrodotoxin resistant sodium channels

Her, brukes en ny teknikk kalt Tetrotoxin (TTX) foretatt motstand for undersøkelser synapser (TERPS) for å teste for monosynaptic tilkoblinger mellom målet neurons. Metoden er avhengig av co uttrykk for en transgene aktivator med tetrodotoxin-resistente natrium kanalen, NaChBac, i en bestemt presynaptic Nevron. Forbindelser med antatte etter synaptic partnere bestemmes av hele celle opptak i nærvær av TTX, som blokkerer elektrisk aktivitet i nerveceller som ikke uttrykker NaChBac. Denne tilnærmingen kan endres for å arbeide med Aktivator eller kalsium bildebehandling som reporter tilkoblinger. TERPS legges til voksende verktøy tilgjengelig for å bestemme tilkobling i nettverk. Men er TERPS unik i at den rapporterer også pålitelig bulk eller volum overføring og smitteeffekter overføring.

TERPS brukes til å skille mellom mono- og polysynaptic bidrag i synaptic tilkoblinger mellom neurons. Mens svak stimulering av en celle kan brukes til å teste direkte forbindelser, rekrutterer kjører større presynaptic aktivitet ofte polysynaptic tilkoblinger (figur 1A). TERPS arbeider av co uttrykke TTX tittelen natrium kanalen NaChBac og optogenetic Aktivator og testing tilkoblinger i nærvær av TTX å eliminere polysynaptic tilkoblinger (<strong class...

Watch this Scientific Journal Video about Examining Monosynaptic Connections in Drosophila Using Tetrodotoxin Resistant Sodium Channels at JoVE.com

Examining Monosynaptic Connections in Drosophila Using Tetrodotoxin Resistant Sodium Channels

Denne artikkelen inneholder en metode for å teste monosynaptic tilkoblinger mellom neurons ved å bruke tetrodotoxin og tetrodotoxin-resistente natrium kanalen, NaChBac.

Undersøke Monosynaptic forbindelser i Drosophila bruke Tetrodotoxin motstandsdyktig natrium kanaler

Here, a new technique termed Tetrotoxin (TTX) Engineered Resistance for Probing Synapses (TERPS) is applied to test for monosynaptic connections between ...

Research

JoVE Journal

Neuroscience

Examining Monosynaptic Connections in Drosophila Using Tetrodotoxin Resistant Sodium Channels

Examining Local Network Processing using Multi-contact Laminar Electrode Recording

Cortical layers are ubiquitous structures throughout neocortex1-4 that consist of highly recurrent local networks. In recent years, significant progress ...

Correlating Behavioral Responses to fMRI Signals from Human Prefrontal Cortex: Examining Cognitive Processes Using Task Analysis

Samkjøre Behavioral Responses til fMRI Signaler fra Menneskelig prefrontal cortex: Undersøke kognitive prosesser Bruke Task Analysis

The aim of this methods paper is to describe how to implement a neuroimaging technique to examine complementary brain processes engaged by two similar ...

Visualization of Proprioceptors in Drosophila Larvae and Pupae

Visualization of Proprioceptors in Drosophila Larvae and Pupae

Visualisering av Proprioceptors i<em&gt; Drosophila</em&gt; Larver og pupper

Proprioception is the ability to sense the motion, or position, of body parts by responding to stimuli arising within the body. In fruitflies and other ...

Multi-photon Intracellular Sodium Imaging Combined with UV-mediated Focal Uncaging of Glutamate in CA1 Pyramidal Neurons

Multi-foton Intracellulær Sodium Imaging Kombinert med UV-mediert Focal Uncaging av Glutamat i CA1 Pyramideformet Neuroner

Multi-photon fluorescence microscopy has enabled the analysis of morphological and physiological parameters of brain cells in the intact tissue with high ...

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

Bruke Lineær Agarose kanaler å studere<em&gt; Drosophila</em&gt; Larve Crawling Behavior

Drosophila larval crawling is emerging as a powerful model to study neural control of sensorimotor behavior. However, larval crawling behavior on flat ...

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

En enkel måte å måle endringer i Reward-søkende atferd med<em&gt; Drosophila melanogaster</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

Quantitative Cell Biology of Neurodegeneration in Drosophila Through Unbiased Analysis of Fluorescently Tagged Proteins Using ImageJ

Quantitative Cell Biology of Neurodegeneration in Drosophila Through Unbiased Analysis of Fluorescently Tagged Proteins Using ImageJ

Kvantitativ cellebiologi mentalt tilbakestående da i Drosophila gjennom objektiv analyse av Fluorescently merket proteiner med ImageJ

With the rising prevalence of neurodegenerative diseases, it is increasingly important to understand the underlying pathophysiology that leads to neuronal ...

Quantification of Macronutrients Intake in a Thermogenetic Neuronal Screen using Drosophila Larvae

Quantification of Macronutrients Intake in a Thermogenetic Neuronal Screen using Drosophila Larvae

Kvantifisering av makronæringsstoffer Inntak i en termogenetisk neuronal skjerm ved hjelp av Drosophila larver

Foraging and feeding behaviors allow animals to access sources of energy and nutrients essential for their development, health, and fitness. Investigating ...

Unraveling the Role of Discrete Areas of the Rat Brain in the Regulation of Ovulation through Reversible Inactivation by Tetrodotoxin Microinjections

Unraveling rollen som diskrete områder av rottehjernen i regulering av eggløsning gjennom reversibel inaktivering av tetrodtoksin mikroinjeksjoner

Many experimental approaches have been used for studying the role of the brain in the regulation of ovulation. Examples include the lesion and ...

A Standardized Pipeline for Examining Human Cerebellar Grey Matter Morphometry using Structural Magnetic Resonance Imaging

En standardisert rørledning for undersøkelse av humant cerebellargrå materiemorfometri ved hjelp av strukturell magnetisk resonansavbildning

Multiple lines of research provide compelling evidence for a role of the cerebellum in a wide array of cognitive and affective functions, going far beyond ...