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Shuttle-box avoidance learning is well-established in behavioral neuroscience. This protocol describes how shuttle-box learning in rodents can be combined with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical in vivo recordings as a tool to study multiple aspects of learning and perception.
Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the necessary equipment is now available by several commercial companies. This protocol provides a detailed description of a two-way shuttle-box avoidance learning paradigm in rodents (here Mongolian gerbils; Meriones unguiculatus) in combination with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical electrophysiological in vivo recordings. The detailed protocol is applicable to study multiple aspects of learning behavior and perception in different rodent species.
Site-specific ICMS of auditory cortical circuits as conditioned stimuli here is used as a tool to test the perceptual relevance of specific afferent, efferent and intracortical connections. Distinct activation patterns can be evoked by using different stimulation electrode arrays for local, layer-dependent ICMS or distant ICMS sites. Utilizing behavioral signal detection analysis it can be determined which stimulation strategy is most effective for eliciting a behaviorally detectable and salient signal. Further, parallel multichannel-recordings using different electrode designs (surface electrodes, depth electrodes, etc.) allow for investigating neuronal observables over the time course of such learning processes. It will be discussed how changes of the behavioral design can increase the cognitive complexity (e.g. detection, discrimination, reversal learning).
A fundamental aim of behavioral neuroscience is to establish specific links between neuronal structural and functional properties, learning, and perception. Neural activity associated with perception and learning can be studied by electrophysiological recording of action potentials and local field potentials in various brain structures at multiple sites. Whereas electrophysiological recordings provide correlative associations between neural activity and behavior, direct electrical intracortical microstimulation (ICMS) for over a century has been the most direct method for testing causal relationships of excited populations of neurons and their behavio....
All experiments presented in this work were conducted in agreement with the ethical standards defined by the German law for the protection of experimental animals. Experiments were approved by the ethics committee of the state of Saxony-Anhalt.
1. Custom-made Multichannel Electrode Arrays for Microstimulation and Recording
This section illustrates a representative example of shuttle-box learning in a Mongolian gerbil. The subject was trained to discriminate the ICMS site between two stimulation electrodes implanted 700 µm apart from each other in auditory cortex (Figures 1 and 2). Stimulation arrays can be customized in different spatial designs (Figure 1). Here, discrimination of the two ICMS sites was learned within 3 training sessions with presentation of 30 CS+ and CS- each (Figure 3A-C
This protocol describes a method of simultaneous site-specific ICMS and multi-channel electrophysiological recordings in a learning animal by using a two-way aversive foot-shock controlled shuttle-box system. The protocol emphasizes technical key concepts for such combination and points out the importance of grounding the animal only via its common ground electrode, leaving the gridfloor at a floating voltage. Here, auditory shuttle-box learning was applied to Mongolian gerbils as learning-related plastic reorganizations.......
The work was supported by grants from the Deustche Forschungsgemeinschaft DFG and the Leibniz-Institute for Neurobiology. We thank Maria-Marina Zempeltzi and Kathrin Ohl for technical assistance.
....Name | Company | Catalog Number | Comments |
Teflon-insulated stainless steel wire | California Fine Wire | diam. 50µm w/ isolation | |
Pin connector system | Molex Holding GmbH | 510470200 | 1.25 mm pitch PicoBlade |
TEM grid Quantifoil | Science Services | EQ225-N27 | |
Dental acrylic Paladur | Heraeus Kulzer | 64707938 | |
Hand-held drill OmniDrill35 | WPIÂ | 503599 | |
Ketamine 500mg/10ml | Ratiopharm GmbH | 7538837 | |
Rompun 2%, 25ml | Bayer Vital GmbH | 5066.0 | |
Sodium-Chloride 0.9%, 10ml | B.Braun AGÂ | PRID00000772 | |
Lubricant KY-Jelly | Johnson & Johnson | ||
Shuttle-box E10-E15 | Coulbourn Instruments | H10-11M-SC | |
Stimulus generator MCS STG 2000 | Multichannel Systems | ||
Plexon Headstage cable 32V-G20 | Plexon Inc. | HSC/32v-G20 | |
Plexon Headstage 32V-G20 | Plexon Inc. | HST/32v-G20 | |
PBX preamplifier 32 channels | Plexon Inc. | 32PBX box | |
Multichannel Acquisition System | Plexon Inc. | MAP 32/HLK2 | |
Cryostate CM3050 S | Leica Microsystems GmbH | ||
Signal processing Card Ni-Daq | National Instruments | ||
Lab StandardTM Stereotaxic Instruments | Stoelting Co. | ||
Audio attenator g.pah | g.pah Guger technologies | ||
Cresyl violet acetate | Roth GmbH | 7651.2 | |
Roticlear | Roth GmbH | A538.1 | |
Sodium acetate trihydrate | Roth GmbH | 6779.1 | |
Potassium hexacyanoferrat(II) trihydrate | Roth GmbH | 7974.2 | |
Di-sodium hydrogen phospahte dihydrate | Merck | 1,065,801,000 | |
ICM Impedance Conditioning Module | FHC | 55-70-0 | |
Animal Temperarture Controler | World Precision Instruments | ATC2000 |
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