Name: acute in vivo electrophysiological recordings of local field potentials and multi-unit activity from the hyperdirect pathway in anesthetized rats
Uploaded: 2017-06-22T15:00:00
Description: Watch this Scientific Journal Video about Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats at JoVE.com

We thank the Deutsche Forschungsgemeinschaft (DFG), KFO 247, for funding our study.

Experimental procedures were conducted in accordance with the German Animal Welfare Act (last revised in 2014) and European regulations (2010/63/EU). Experiments were approved by the local animal welfare authority (LaGeSo, Berlin), and conformed to local department and international guidelines.
NOTE: In the presented method two models of electrodes are used to record from the hyperdirect cortico-basal ganglia pathway that connects the primary motor cortex (M1) with the subthalamic nucleus (STN...

<ol>
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C., Poloskey, S. L., Bergstrom, D. A., Walters, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parafascicular+thalamic+nucleus+activity+in+a+rat+model+of+Parkinson's+disease.">Parafascicular thalamic nucleus activity in a rat model of Parkinson's disease.</a> Exp Neurol. 217 (2), 269-281 (2009).</li><li>Steriade, M., Nunez, A., Amzica, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+slow+(&lt;+1+Hz)+oscillation+of+neocortical+neurons+in+vivo:+depolarizing+and+hyperpolarizing+components.">A novel slow (&lt; 1 Hz) oscillation of neocortical neurons in vivo: depolarizing and hyperpolarizing components.</a> J Neurosci. 13 (8), 3252-3265 (1993).</li><li>Maggi, C. A., Meli, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suitability+of+urethane+anesthesia+for+physiopharmacological+investigations+in+various+systems.+Part+1:+General+considerations.">Suitability of urethane anesthesia for physiopharmacological investigations in various systems. Part 1: General considerations.</a> Experientia. 42 (2), 109-114 (1986).</li><li>Goldberg, J. A., Kats, S. S., Jaeger, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Globus+pallidus+discharge+is+coincident+with+striatal+activity+during+global+slow+wave+activity+in+the+rat.">Globus pallidus discharge is coincident with striatal activity during global slow wave activity in the rat.</a> J Neurosci. 23 (31), 10058-10063 (2003).</li><li>Karain, B., Xu, D., Bellone, J. A., Hartman, R. E., Shi, W. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+globus+pallidus+neurons:+functional+classification+and+effects+of+dopamine+depletion.">Rat globus pallidus neurons: functional classification and effects of dopamine depletion.</a> Synapse. 69 (1), 41-51 (2015).</li><li>Paasonen, 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=Comparison+of+seven+different+anesthesia+protocols+for+nicotine+pharmacologic+magnetic+resonance+imaging+in+rat.">Comparison of seven different anesthesia protocols for nicotine pharmacologic magnetic resonance imaging in rat.</a> Eur Neuropsychopharmacol. 26 (3), 518-531 (2016).</li><li>Mahmud, M., Vassanelli, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Processing+and+Analysis+of+Multichannel+Extracellular+Neuronal+Signals:+State-of-the-Art+and+Challenges.">Processing and Analysis of Multichannel Extracellular Neuronal Signals: State-of-the-Art and Challenges.</a> Front Neurosci. 10, 248 (2016).</li><li>Hadar, R., 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=Altered+neural+oscillations+and+elevated+dopamine+levels+in+the+reward+pathway+during+alcohol+relapse.">Altered neural oscillations and elevated dopamine levels in the reward pathway during alcohol relapse.</a> Behav Brain Res. 316, 131-135 (2017).</li><li>Voget, M., 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=Altered+local+field+potential+activity+and+serotonergic+neurotransmission+are+further+characteristics+of+the+Flinders+sensitive+line+rat+model+of+depression.">Altered local field potential activity and serotonergic neurotransmission are further characteristics of the Flinders sensitive line rat model of depression.</a> Behav Brain Res. 291, 299-305 (2015).</li></ol>

In the present study, the method is demonstrated how to record extracellular electrophysiological signals simultaneously from multiple sites of a given network using the example of the hyperdirect cortico-basal ganglia pathway that connects the M1 with the STN and SNr in rodents.
A critical step in the recording of small subcortical structures such as the STN is the precisely guided insertion of the recording electrodes into the target. In the presented method, taking care of two crucial steps...

Recent cumulative evidence on different neuropsychiatric disorders such as Parkinson&#180;s disease (PD) and schizophrenia strongly suggests that their pathophysiology is based on a critical dysfunction of extended neuronal circuits that often involve cortical and subcortical structures1,2,3. According to this theory, the clinical manifestations of the diseases arise as a consequence of an impaired information processing capacity of a network of cells instead of single cells or specific neuronal elements1,2,3. In order to enhance the understanding of this complex group of neuropsychiatric diseases and to find new treatment options, it is mandatory to characterize the neuronal dynamics of those disordered networks in human patients and in animal models in great detail. An excellent method to study large-scale networks in living subjects is multi-site electrophysiological recordings of extracellular potentials4. Using this method, it is possible to simultaneously assess local field potentials (LFPs), that primarily represent the temporal summation of excitatory and inhibitory postsynaptic currents and multi-unit activity (MUA), that is generated by presynaptic potentials5. The recording of extracellular potentials has several advantages over alternative methods to study networks, e.g., functional magnetic resonance imaging and calcium imaging, because it provides a higher temporal and spatial resolution and because it is not dependent on genetically engineered organisms5. However, the use of extracellular in vivo recordings is limited since it is an invasive technique that cannot be universally applied.
In vivo electrophysiological recordings can be performed in awake as well as in anesthetized animals6. Both methods are accompanied by specific pros and cons. Studies in awake animals allow the recording of brain signals during the performance of defined behavioral tasks, but are prone to movement related and other artifacts7,8. Recordings in anesthetized animals on the other hand offer the opportunity to assess LFPs and MUA with a minimum of artifacts at highly defined cortical synchronization states, but the results also differ to some extent to what can be found in awake subjects9,10,11.
In recent years, it has been demonstrated that the sampling of LFPs is especially useful to delineate pathological changes of network activity. A prominent example of this is research on the pathophysiology of PD in human patients and animal models of the disease, where it could be shown that enhanced beta oscillations in the cortico-basal ganglia loop are linked to parkinsonian motor symptoms12,13. As a consequence of this line of research, it is currently investigated if beta oscillations could be used as an online feedback biomarker for closed-loop deep brain stimulation14,15.
In the present study, a detailed description of acute multi-site in vivo electrophysiological recordings of LFPs and MUA in rats anesthetized with urethane is provided. It is demonstrated how a complete network, such as the hyperdirect cortico-basal ganglia pathway can be characterized electrophysiologically using standard and customized electrodes and how those electrodes can be build. It is especially emphasized how a precise targeting of basal ganglia nuclei can be achieved by combining stereotactic surgery together with the online registration of MUAs.

<table>
	<tbody>
		<tr>
			<td>Ag/AgCl custom epidural electrodes</td>
			<td>Goodfellow GmbH 
			D-61213 Bad Nauheim, Germany 
			info@goodfellow.com</td>
			<td>Product-ID AG005127 for 99.99% silver wire</td>
			<td>Ag/AgCl electrodes will allow for better signal quality, but may only be used in acute experiments. Possible replacement: Stainless steel electrodes</td>
		</tr>
		<tr>
			<td>Stereotaxic holder with acrylic block</td>
			<td>David Kopf Instruments, 
			7324 Elmo Street, Tujunga, CA 91042, USA</td>
			<td>Product ID Model 1770 Standard Electrode Holder</td>
			<td>Make sure the acrylic block has recesses which suit the electrode setup for the desired target. Acrylic blocks can easily be modified with a file to obtain the desired configuration. Possible replacement: Self-constructed electrode holders</td>
		</tr>
		<tr>
			<td>Tungsten microwire electrodes 1.5 M&#937; impedance</td>
			<td>Microprobes.com 
			18247-D Flower Hill Way&#160; Gaithersburg, Maryland, 20879 USA</td>
			<td>Product-ID WE3ST31.5A5-250um</td>
			<td>The 1.5 M&#937; is necessary to record MUA and LFP at the same time. Possible replacement: Microelectrodes of different materials can be used. The electrodes have to be straight, robust and as thin as possible.</td>
		</tr>
		<tr>
			<td>Rat alignment tool</td>
			<td>David Kopf Instruments, 
			7324 Elmo Street, Tujunga, CA 91042, USA</td>
			<td>Product ID Model 944 Rat Alignment Tool</td>
			<td>Allows the exact orientation of the brain to match stereotaxic atlases. Possible replacement: Stereotaxic holder with a cannula</td>
		</tr>
		<tr>
			<td>Two-component dental acrylic</td>
			<td>Associated Dental Products Ltd. 
			Kemdent Works, Purton, Swindon 
			Wiltshire, SN5 4HT, United Kingdom</td>
			<td>Simplex Rapid Powder Clear 225g, Product code: ACR803; Simplex Rapid Liquid 150ml, Product code: ACR920</td>
			<td>Depending in the electrodes used, superglue might be an easy alternative, if the electrodes are small and lightweight. Possible replacement: Superglue (Cyanacrylate-based)</td>
		</tr>
		<tr>
			<td>Faraday cage</td>
			<td>Self-construction</td>
			<td></td>
			<td>A proper Faraday cage will be the best protection from electromagnetic artifacts, but everything which can be formed into a box shape or applied to a frame and is made of conductive material may help. Possible replacement: Aluminum foil or copper mesh</td>
		</tr>
		<tr>
			<td>Electrophysiological setup with recording software and online spike-sorting capabilities</td>
			<td>OmniPlex&#174; Neural Data Acquisition System 
			Plexon Inc 
			6500 Greenville Avenue, Suite 700 
			Dallas, Texas 75206 
			USA</td>
			<td></td>
			<td>Offline sorting software is a potential alternative, multiple scripts and softwares can be found for free in the open source community.</td>
		</tr>
	</tbody>
</table>

acute in vivo electrophysiological recordings of local field potentials and multi-unit activity from the hyperdirect pathway in anesthetized rats

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the pathophysiological basis of these diseases, it is necessary to precisely characterize in which way the processing of information is disturbed between the different neuronal parts of the circuit. Using extracellular in vivo electrophysiological recordings, it is possible to accurately delineate neuronal activity within a neuronal network. The application of this method has several advantages over alternative techniques, e.g., functional magnetic resonance imaging and calcium imaging, as it allows a unique temporal and spatial resolution and does not rely on genetically engineered organisms. However, the use of extracellular in vivo recordings is limited since it is an invasive technique that cannot be universally applied. In this article, a simple and easy to use method is presented with which it is possible to simultaneously record extracellular potentials such as local field potentials and multiunit activity at multiple sites of a network. It is detailed how a precise targeting of subcortical nuclei can be achieved using a combination of stereotactic surgery and online analysis of multi-unit recordings. Thus, it is demonstrated, how a complete network such as the hyperdirect cortico-basal ganglia loop can be studied in anesthetized animals in vivo.

With the herein used recording electrodes, it is possible to sample LFPs from the primary motor cortex, the subthalamic nucleus and the substantia nigra pars reticulata and MUA from the STN and SNr. Initially, LFPs and multi-unit activity are recorded together in a broad-band signal. Thereafter, LFPs and MUAs are separated by bandpass filters (0.05-250 Hz for LFPs and 300-4,000 Hz for MUA).
For the correct targeting of subcortic...

Watch this Scientific Journal Video about Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats at JoVE.com

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

In this study, the methodology is presented on how to perform multi-site in vivo electrophysiological recordings from the hyperdirect pathway under urethane anesthesia.

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the ...

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