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 Video at JoVE.com

Cette recherche a été financée par le NIH et la NSF subvention NS047073 subvention DMS1010434.

1. Préparation des solutions <ol><li> À préparer une solution de chlorure de magnésium qui est isotonique à l&#39;eau de mer dans laquelle les animaux sont gardés (~ 1.000 millosmolar), marquer un grand bol au niveau du volume souhaité. Remplissez le pot avec de l&#39;eau distillée à environ 80% de ce niveau, et pesez la quantité appropriée de chlorure de magnésium hexahydraté pour créer une solution de 333 mm dans le volume final. Ajouter le chlorure de magnésium à la cruche, fermer le couvercle et ag...

<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. W., Chiel, H. J., Susswein, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activity+patterns+of+the+B31/B32+pattern+initiators+innervating+the+I2+muscle+of+the+buccal+mass+during+normal+feeding+movements+in+Aplysia+californica.">Activity patterns of the B31/B32 pattern initiators innervating the I2 muscle of the buccal mass during normal feeding movements in Aplysia californica.</a> J. Neurophys. 75, 1309-1326 (1996).</li><li>Cullins, M. J., 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=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.</a> J. Vis. Exp. (40), e1791 (2010).</li><li>Warman, E. N., 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=A+new+technique+for+chronic+single+extracellular+recording+in+freely+behaving+animals+using+pipette+electrodes.">A new technique for chronic single extracellular recording in freely behaving animals using pipette electrodes.</a> J. Neurosci. Methods. 57, 161-169 (1995).</li><li>Lu, H., Chestek, C. A., Shaw, K. M., 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=Selective+extracellular+stimulation+of+individual+neurons+in+ganglia.">Selective extracellular stimulation of individual neurons in ganglia.</a> J. Neural Eng. 5, 287-309 (2008).</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=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>

Des travaux antérieurs ont caractérisé l&#39;aplysie programmes moteurs chez l&#39;animal intact et dans la préparation réduits, tels que les ganglions isolé. Chez l&#39;animal intact, bien que des enregistrements de neurones individuels ont été obtenus 5, de telles expériences sont très difficiles, et les électrodes ne peut pas être déplacé d&#39;un neurone à pendant l&#39;alimentation. Dans les ganglions isolés, les mouvements alimentation induites par l&#39;activité neuronale peut...

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	<tbody>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<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>
		</tr>
		<tr>
			<td>Carbachol</td>
			<td>Acros Organics</td>
			<td>10824</td>
			<td>99%</td>
		</tr>
		<tr>
			<td>Sodium hydroxide</td>
			<td>Fisher Scientific</td>
			<td>SS255</td>
			<td>Certified</td>
		</tr>
		<tr>
			<td>Hydrochloric acid</td>
			<td>Fisher Scientific</td>
			<td>SA49</td>
			<td>Certified</td>
		</tr>
		<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>
		</tr>
		<tr>
			<td>Duro Quick-Gel superglue</td>
			<td>Henkel corp.</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<tr>
			<td>100 x 50 mm Crystalizing Dish</td>
			<td>Pyrex</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>High Vacuum Grease</td>
			<td>Dow Corning</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Pipet Tips</td>
			<td>Fisher Scientific</td>
			<td>21-375D</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Minutien Pins</td>
			<td>Fine Science Tools</td>
			<td>26002-10</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Modeling Clay</td>
			<td>Sargent Art</td>
			<td>22-4400</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Silk Sutures</td>
			<td>Ethicon</td>
			<td>K89OH</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<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

Multifonctionnalité, la capacité d&#39;une structure périphérique de générer plusieurs comportements distincts, 1, permet aux animaux de s&#39;adapter rapidement leurs comportements à des environnements changeants. Le mollusque marin Aplysia californica fournit un système souple pour l&#39;étude de la multifonctionnalité. Au cours de l&#39;alimentation, l&#39;aplysie génère plusieurs types distincts de comportements en utilisant l&#39;appareil d&#39;alimentation même, la masse buccale. Les ganglions qui contrôlent ces comportements contiennent un certain nombre de grands neurones identifiés qui sont accessibles à l&#39;étude électrophysiologique. L&#39;activité de ces neurones a été décrit dans les programmes moteurs qui peuvent être divisés en deux types, les programmes et ingestifs egestive, basé sur le timing de l&#39;activité neuronale qui ferme la pince par rapport à la nourriture de l&#39;activité neuronale qui prolonge ou se rétracte la pince 2. Toutefois, dans les ganglions isolés, les mouvements musculaires qui produisent ces comportements sont absents, ce qui en faitplus difficile d&#39;être certain que les programmes moteurs observés sont des corrélats des comportements réels. Dans enregistrements in vivo, nerveuses et musculaires ont été obtenus correspondant à des programmes d&#39;alimentation 2,3,4, mais il est très difficile d&#39;enregistrer directement à partir de neurones individuels 5. De plus, in vivo, les programmes ingestifs peut être divisée en morsures et 1,2 hirondelles, une distinction est difficile à faire dans la plupart décrite précédemment préparations in vitro. La préparation de la masse suspendue buccale (figure 1) comble le fossé entre les noyaux isolés et les animaux intacts. Dans cette préparation, les comportements ingestifs - y compris à la fois mordant et avaler - et des comportements egestive (rejet) peut être obtenue, en même temps que des neurones individuels peuvent être enregistrées et à partir de l&#39;utilisation d&#39;électrodes stimulées extracellulaires 6. Les mouvements d&#39;alimentation associés à ces comportements différents peuvent être enreded, quantifiés et directement liés aux programmes moteurs. Les programmes moteurs dans la préparation de la masse suspendue buccale semblent être plus semblables à ceux observés in vivo que sont les programmes moteurs suscité dans les ganglions isolé. Ainsi, les programmes moteurs de cette préparation peut être plus directement lié au comportement in vivo; dans le même temps, les neurones individuels sont plus accessibles à l&#39;enregistrement et la stimulation que chez les animaux intacts. En outre, comme une étape intermédiaire entre les noyaux isolés et les animaux intacts, les conclusions de la masse suspendue buccale peut faciliter l&#39;interprétation des données obtenues dans des contextes à la fois plus réduites et plus intacte. La préparation de la masse suspendue buccale est un outil utile pour caractériser le contrôle neural de la multifonctionnalité de l&#39;aplysie.

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 Video at JoVE.com

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

Nous décrivons une technique d&#39;enregistrement extracellulaire et stimuler les nerfs, les muscles et les neurones individuels identifiés<em&gt; In vitro</em&gt; Tout en recueillant et en observant les différents types de comportements alimentaires dans le dispositif d&#39;alimentation de<em&gt; Aplysie</em&gt;.

Un<em&gt; In Vitro</em&gt; Préparation Susciter et enregistrement des programmes d&#39;alimentation moteur avec les mouvements physiologiques en<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

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Fabrication des électrodes et l&#39;implantation de Aplysia californica Des enregistrements multi-canaux neuronaux et musculaires au Intact, Animaux Librement Behaving

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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Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo Video (Video) | JoVE

Méthode électrophysiologique de réponses d&#39;enregistrement intracellulaire tension de<em&gt; Drosophila</em&gt; Photorécepteurs et Interneurones à Light Stimuli<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

Une seule étape simple protocole Dissection pour Whole-mount Préparation des adultes<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

Préparations et protocoles pour toute cellule Patch Clamp enregistrement des neurones Tectal Xenopus laevis

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