Name: an in vitro preparation for eliciting and recording feeding motor programs with physiological movements in aplysia californica
Uploaded: 2012-12-05T13:00:00
Description: Watch this Scientific Journal Video about An In Vitro Preparation for Eliciting and Recording Feeding Motor Programs with Physiological Movements in Aplysia californica at JoVE.com

Denna forskning stöddes av NIH bidrag NS047073 och NSF bevilja DMS1010434.

1. Framställning av lösningar <ol><li> För att bereda lösning magnesiumklorid som är isoton med havsvatten, i vilken djuren hålls (~ 1.000 millosmolar), markera en stor kanna i nivå med önskad volym. Fyll kannan med destillerat vatten till ungefär 80% av denna nivå, och väga den lämpliga mängden av magnesiumklorid-hexahydrat för att skapa en 333 mM lösning i den slutliga volymen. Lägg till magnesiumklorid till kannan, stäng locket och skaka kraftigt tills magnesiumklorid är helt upplöst, och sedan l...

<ol>
	<li>Neustadter, D. M., Herman, R. L., Drushel, R. F., Chestek, D. W., Chiel, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+kinematics+of+multifunctionality:+comparisons+of+biting+and+swallowing+in+Aplysia+californica.">The kinematics of multifunctionality: comparisons of biting and swallowing in Aplysia californica.</a> J. Exp. Biol. 210, 238-260 (2007).</li><li>Morton, D. W., Chiel, H. J. <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+buccal+nerve+activity+that+distinguishes+ingestion+from+rejection+can+be+used+to+predict+behavioral+transitions+in+Aplysia.">In vivo buccal nerve activity that distinguishes ingestion from rejection can be used to predict behavioral transitions in Aplysia.</a> J. Comp. Physiol. A. 172, 17-32 (1993).</li><li>Hurwitz, I., Neustadter, D., Morton, D. 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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+diverse+neuropeptide+cotransmitters+by+identified+motor+neurons+in+Aplysia.">Expression of diverse neuropeptide cotransmitters by identified motor neurons in Aplysia.</a> J. Neurosci. 11, 618-625 (1991).</li><li>Church, P. J., Lloyd, P. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activity+of+multiple+identified+motor+neurons+recorded+intracellularly+during+evoked+feedinglike+motor+programs+in+Aplysia.">Activity of multiple identified motor neurons recorded intracellularly during evoked feedinglike motor programs in Aplysia.</a> J. Neurophys. 72, 1794-1809 (1994).</li><li>Nargeot, R. N., Baxter, D. A., Byrne, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contingent-dependent+enhancement+of+rhythmic+motor+patterns:+an+in+vitro+analog+of+operant+conditioning.">Contingent-dependent enhancement of rhythmic motor patterns: an in vitro analog of operant conditioning.</a> J. Neurosci. 17, 8093-8105 (1997).</li><li>Kandel, E. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Behavioral biology of Aplysia. , (1979).</li><li>Scott, M. L., Govind, C. K., Kirk, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuromuscular+organization+of+the+buccal+system+in+Aplysia+californica.">Neuromuscular organization of the buccal system in Aplysia californica.</a> J. Comp. Neurol. 312, 207-222 (1991).</li><li>Susswein, A. J., Rosen, S. C., Gapon, S., Kupfermann, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+buccal+motor+programs+elicited+by+a+cholinergic+agonist+applied+to+the+cerebral+ganglion+of+Aplysia+californica.">Characterization of buccal motor programs elicited by a cholinergic agonist applied to the cerebral ganglion of Aplysia californica.</a> J. Comp. Physiol. A. 179, 509-524 (1996).</li><li>Kupfermann, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feeding+behavior+in+Aplysia:+A+simple+system+for+the+study+of+motivation.">Feeding behavior in Aplysia: A simple system for the study of motivation.</a> Behav. Biol. 10, 1-26 (1974).</li><li>Morton, D. W., Chiel, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+timing+of+activity+in+motor+neurons+that+produce+radula+movements+distinguishes+ingestion+from+rejection+in+Aplysia.">The timing of activity in motor neurons that produce radula movements distinguishes ingestion from rejection in Aplysia.</a> J. Comp. Physiol. A. 173, 519-536 (1993).</li><li>Weiss, K. R., Chiel, H. J., Koch, U., Kupfermann, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activity+of+an+identified+histaminergic+neuron,+and+its+possible+role+in+arousal+of+feeding+behavior+in+semi-intact+Aplysia.">Activity of an identified histaminergic neuron, and its possible role in arousal of feeding behavior in semi-intact Aplysia.</a> J. Neurosci. 6, 2403-2415 (1986).</li><li>Jing, J., Weiss, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+variants+of+a+motor+act+in+a+modular+and+hierarchical+motor+network.">Generation of variants of a motor act in a modular and hierarchical motor network.</a> Curr. Biol. 15, 1712-1721 (2005).</li><li>Jing, J., Weiss, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+mechanisms+of+motor+program+switching+in+Aplysia.">Neural mechanisms of motor program switching in Aplysia.</a> J. Neurosci. 21, 7349-7362 (2001).</li><li>Morgan, P. T., Jing, J., Vilim, F. S., Weiss, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interneuronal+and+peptidergic+control+of+motor+pattern+switching+in+Aplysia.">Interneuronal and peptidergic control of motor pattern switching in Aplysia.</a> J. Neurophysiol. 87, 49-61 (2002).</li><li>Jing, J., Cropper, E. C., Hurwitz, I., Weiss, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+construction+of+movement+with+behavior-specific+and+behavior-independent+modules.">The construction of movement with behavior-specific and behavior-independent modules.</a> J. Neurosci. 24, 6315-6325 (2004).</li></ol>

Tidigare arbete har präglat Aplysia motor program i intakta djur och i minskade preparat, såsom isolerad ganglier. I det intakta djuret, även inspelningar av enskilda neuroner har erhållits 5, sådana experiment är mycket svåra, och elektroder kan inte flyttas från neuron till neuron under matning. I isolerade ganglier kan utfodring rörelser som induceras av neural aktivitet inte observeras. Den suspenderade buckala massa förberedelse överbryggar gapet mellan dessa två ytterligheter. <...

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			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
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			<td>Sodium chloride</td>
			<td>Fisher Scientific</td>
			<td>S671</td>
			<td>Biological, Certified</td>
		</tr>
		<tr>
			<td>Potassium chloride</td>
			<td>Fisher Scientific</td>
			<td>P217</td>
			<td>Certified ACS</td>
		</tr>
		<tr>
			<td>Magnesium chloride hexahydrate</td>
			<td>Acros Organics</td>
			<td>19753</td>
			<td>99%</td>
		</tr>
		<tr>
			<td>Magnesium sulfate heptahydrate</td>
			<td>Fisher Scientific</td>
			<td>M63</td>
			<td>Certified ACS</td>
		</tr>
		<tr>
			<td>Calcium chloride dihydrate</td>
			<td>Fisher Scientifc</td>
			<td>C79</td>
			<td>Certified ACS</td>
		</tr>
		<tr>
			<td>Glucose (dextrose)</td>
			<td>Sigma-Aldrich</td>
			<td>G7528</td>
			<td>BioXtra</td>
		</tr>
		<tr>
			<td>MOPS buffer</td>
			<td>Acros Organics</td>
			<td>17263</td>
			<td>99%</td>
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			<td>Carbachol</td>
			<td>Acros Organics</td>
			<td>10824</td>
			<td>99%</td>
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			<td>Sodium hydroxide</td>
			<td>Fisher Scientific</td>
			<td>SS255</td>
			<td>Certified</td>
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			<td>Hydrochloric acid</td>
			<td>Fisher Scientific</td>
			<td>SA49</td>
			<td>Certified</td>
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		<tr>
			<td>Single-barreled capillary glass</td>
			<td>A-M Systems</td>
			<td>6150</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Flaming-Brown micropipette puller model P-80/PC</td>
			<td>Sutter Instruments</td>
			<td>&nbsp;</td>
			<td>Filament used: FT345B</td>
		</tr>
		<tr>
			<td>Enamel coated stainless steel wire</td>
			<td>California Fine Wire</td>
			<td>&nbsp;</td>
			<td>0.001D, coating h</td>
		</tr>
		<tr>
			<td>Household Silicone II Glue</td>
			<td>GE</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
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			<td>Duro Quick-Gel superglue</td>
			<td>Henkel corp.</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
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		<tr>
			<td>A-M Systems model 1700 amplifier</td>
			<td>A-M Systems</td>
			<td>&nbsp;</td>
			<td>Filter settings: 300-500 Hz nerves,10-500 Hz I2 muscle</td>
		</tr>
		<tr>
			<td>Pulsemaster Multi-Channel Stimulator</td>
			<td>World Precision Instruments</td>
			<td>A300</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Stimulus Isolator</td>
			<td>World Precision Instruments</td>
			<td>A360</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>AxoGraph X</td>
			<td>AxoGraph Scientific</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Veeder-Root Totalizing Counter</td>
			<td>Danaher</td>
			<td>C342-0562</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Gold Connector Pins</td>
			<td>Bulgin</td>
			<td>SA3148/1</td>
			<td>&nbsp;</td>
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		<tr>
			<td>Gold Connector Sockets</td>
			<td>Bulgin</td>
			<td>SA3149/1</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Sylgard 184 Silicone Elastomer</td>
			<td>Dow Corning</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
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			<td>100 x 50 mm Crystalizing Dish</td>
			<td>Pyrex</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
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		<tr>
			<td>High Vacuum Grease</td>
			<td>Dow Corning</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
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			<td>Pipet Tips</td>
			<td>Fisher Scientific</td>
			<td>21-375D</td>
			<td>&nbsp;</td>
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		<tr>
			<td>Minutien Pins</td>
			<td>Fine Science Tools</td>
			<td>26002-10</td>
			<td>&nbsp;</td>
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			<td>Modeling Clay</td>
			<td>Sargent Art</td>
			<td>22-4400</td>
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			<td>Silk Sutures</td>
			<td>Ethicon</td>
			<td>K89OH</td>
			<td>&nbsp;</td>
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			<td>Whisper Air Pump</td>
			<td>Tetra</td>
			<td>77849</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Aquarium Tubing</td>
			<td>Eheim</td>
			<td>7783</td>
			<td>12/16 mm</td>
		</tr>
		<tr>
			<td>Elite Airstone</td>
			<td>Hagen</td>
			<td>A962</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Vannas Spring Scissors</td>
			<td>Fine Science Tools</td>
			<td>15000-08</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Dumont #5 Fine Forceps</td>
			<td>Fine Science Tools</td>
			<td>11254-20</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Yaki Sushi Nori Seaweed</td>
			<td>Rhee Bros</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Kimwipes</td>
			<td>Kimberly-Clark</td>
			<td>34155</td>
			<td>&nbsp;</td>
		</tr>
	</tbody>
</table>

an in vitro preparation for eliciting and recording feeding motor programs with physiological movements in aplysia californica

Multifunktionalitet, förmåga en perifer struktur för att generera flera, distinkta beteenden 1, tillåter djur snabbt anpassa sina beteenden till föränderliga miljöer. Den marina mollusk Aplysia californica ger en lätthanterlig system för studier av multifunktionalitet. Under utfodring genererar Aplysia flera olika typer av beteenden som använder samma matningsanordning, den buckala massan. Den ganglierna som styr dessa beteenden innehåller ett antal stora identifierade, neuroner som är tillgängliga för elektrofysiologiska studier. Aktiviteten hos dessa nervceller har beskrivits i motor program som kan delas in i två typer, ingestive och egestive program, baserat på tidpunkten för neural aktivitet som stänger den relativa mat griparmen till den neurala aktivitet som protracts eller drar tillbaka griparmen 2. Men i isolerad ganglier de muskelrörelser som skulle ge dessa beteenden är frånvarande, vilket gör detsvårare att vara säker på huruvida motorns observerade program är korrelat riktiga beteenden. In vivo, nerv och muskel inspelningar har erhållits motsvarande matning program 2,3,4, men det är mycket svårt att direkt spela in från enskilda neuroner 5. Dessutom in vivo kan ingestive program delas in ytterligare bett och sväljer 1,2, en skillnad som är svår att göra i de flesta tidigare beskrivits in vitro preparat. Den suspenderade buckala massa preparat (figur 1) överbryggar klyftan mellan isolerade ganglier och intakta djur. I denna beredning ingestive beteenden - inklusive både bita och svälja - kan och egestive beteenden (avvisande) framkallas, samtidigt som enskilda nervceller kan spelas in från och stimuleras att använda extracellulära elektroder 6. Utfodring rörelser i samband med dessa olika beteenden kan RECORded, kvantifieras och direkt relaterade till motorns program. Motorn programmen i suspenderade buckala massa förberedelse verkar vara mer lika dem som observerats in vivo än är motor-program framkallas i isolerade ganglier. Sålunda kan motorn program i denna beredning är mer direkt relaterade till uppträdande in vivo, samtidigt som de enskilda neuroner är mer tillgängliga för inspelning och stimulering än hos intakta djur. Dessutom, som ett mellansteg mellan isolerade ganglier och intakta djur, resultaten från den suspenderade buckala massan kan hjälpa tolkning av data som erhållits i både mer reducerad och mer intakta inställningar. Den suspenderade buckala massa preparat är ett användbart verktyg för att karakterisera neurala kontrollen av multifunktionalitet i Aplysia.

When an extracellular electrode is positioned above a neuron's soma and used to stimulate the neuron, a one-for-one correspondence between spikes on the soma channel and on the nerve(s) the neuron projects to can be observed (Figure 6, left panel, stimulation of identified neuron B9). The soma channel (top channel) is set to stimulating mode when the current is applied (time 1 in the figure), and is then quickly switched to recording mode (time 2). By maintaining the position of the electrode, the...

Watch this Scientific Journal Video about An In Vitro Preparation for Eliciting and Recording Feeding Motor Programs with Physiological Movements in Aplysia californica at JoVE.com 

An In Vitro Preparation for Eliciting and Recording Feeding Motor Programs with Physiological Movements in Aplysia californica (Video) | JoVE 

Vi beskriver en teknik för att extracellulärt registrera och stimulera från nerver, muskler och individuellt identifierade nervceller<em&gt; In vitro</em&gt; Medan framkalla och observera olika typer av utfodring beteenden vid utfodring apparaten<em&gt; Aplysia</em&gt;.

En<em&gt; In Vitro</em&gt; Förberedelser för att framkalla och spela in program Feeding Motor med fysiologiska Rörelser i<em&gt; Aplysia californica</em

Multifunctionality, the ability of one peripheral  structure to generate multiple, distinct behaviors1, allows animals  to rapidly adapt their behaviors ...

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An In Vitro Preparation for Eliciting and Recording Feeding Motor Programs with Physiological Movements in Aplysia californica

Electrode Fabrication and Implantation in Aplysia californica for Multi-channel Neural and Muscular Recordings in Intact, Freely Behaving Animals

Electrode Fabrication and Implantation in Aplysia californica for Multi-channel Neural and Muscular Recordings in Intact, Freely Behaving Animals (Video) | JoVE 

Elektrod Fabrication och implantation i Aplysia californica För flera kanaler Neural och muskulös Inspelningar i intakt, fritt bete Djur

Recording from key nerves and muscles of Aplysia during feeding behavior allows us to study the patterns of neural control in an intact animal.  ...

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Precise patterns of division, migration and differentiation of neural progenitor cells are crucial for proper brain development and function1,2. To ...

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The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

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Registrering och analys av dygnsrytmen i Running-wheel aktivitet hos gnagare

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica

Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica (Video) | JoVE 

Extracellulärt Identifiera motoriska nervceller för en muskel Motor Pool i<em&gt; Aplysia californica</em

In animals with large  identified neurons (e.g. mollusks), analysis of motor pools is done using  intracellular techniques1,2,3,4. Recently,  we developed ...

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Isolation of Sensory Neurons of Aplysia californica for Patch Clamp Recordings of Glutamatergic Currents Video (Video) | JoVE

Isolering av sensoriska neuroner av<em&gt; Aplysia californica</em&gt; För Patch Clamp Inspelningar av glutamaterg Currents

The marine gastropod mollusk Aplysia californica has a venerable  history as a model of nervous system function, with particular significance in  studies ...

Functional Analysis of the Larval Feeding Circuit in Drosophila

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Funktionell analys av Larv Feeding Circuit i Drosophila</i

The serotonergic  feeding circuit in Drosophila  melanogaster larvae can be used to investigate neuronal substrates of  critical importance during the ...

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Elektro Metod för inspelning Intracellulärt Spännings svar av<em&gt; Drosophila</em&gt; Fotoreceptorer och Intern på ljusstimuli<em&gt; In Vivo</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains Video (Video) | JoVE

En enkel steg Dissection protokoll för hel-mount Framställning av Adult<em&gt; Drosophila</em&gt; Brains

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the ...

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons Video (Video) | JoVE

Preparat och protokoll för hela cellen Patch Clamp inspelningen av Xenopus laevis Tectal nervceller

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic ...