recording of local field potential from both hemispheres of the mouse brain

Recording of Local Field Potential from Both Hemispheres of the Mouse Brain

Prior to surgery, confirm the depth of anesthesia of the mouse by performing a tail or toe pinch with forceps. Next, position the mouse in a stereotaxic apparatus, and fix its head. Apply ointment on both eyes to keep them moist.
Then, shave the head and sterilize the area. Make a small incision of 12 to 15 millimeters in the middle of the shaved area. Using forceps, gently pull the scalp away from the midline. Afterward, separate the skin gently, and remove the residual tissue.
Clean the skull using hydrogen peroxide-coated cotton buds. Under a stereomicroscope, drill two small holes of 1- to 1.5-millimeter radii on both the left and right sides of the skull to allow insertion of the recording microelectrodes into the M2 regions.
Carefully, remove the dura mater with a tungsten needle, then, insert two separate recording microelectrodes filled with 0.5 molar sodium chloride into the holes at an angle of 60 degrees using mechanical micromanipulators.
For LFP recording, slowly lower the left and right glass electrodes to the M2 coordinates. For quality control, test the resistance of each electrode using the differential amplifier. Next, set the recording process at 0.1 Hertz high-pass and 1,000 Hertz low-pass with 1,000 times amplification. Collect the digitized raw LFP data in stable state for at least 60 seconds, with the mouse breathing evenly at two breaths per second under anesthesia.

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Nanyang Technological University

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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Animal anesthesia and surgery
<ol>
	<li>Weigh and anesthetize the mouse with your approved anesthesia regimen from your local animal care committee.</li>
	<li>Perform a tail or toe pinch with forceps to confirm deep anesthesia before surgery.</li>
	<li>Position the mouse in a stereotaxic apparatus and fix its head.</li>
	<li>Apply eye ointment on both eyes to keep them moist. Follow your local animal care guidelines regarding pre- and postoperative analgesia.</li>
	<li>Shave the hair using surgical clippers. Make a small incision (12-15 mm) in the middle of the exposed surgical area with scissors. Using forceps, gently pull the scalp away from the midline.</li>
	<li>Separate the skin gently and remove residual tissue. Clean the skull using hydrogen peroxide-coated cotton buds.</li>
	<li>Drill two small holes of radii 1.0-1.5 mm on both the skull&#39;s left and right sides to allow the recording microelectrodes to be inserted into the M2 regions under a stereomicroscope (Figure 1A). 
	NOTE: Stereotaxic locations of bilateral M2: 1.94 mm anterior to the bregma, 1.0 mm lateral to the midline, and 0.8-1.1 mm ventral to the dura.</li>
	<li>Remove the dura mater carefully with a tungsten needle.</li>
	<li>Pull glass borosilicate micropipettes (outer diameter: 1.0 mm) as recording microelectrodes with resistance of 1-2 M&#937;.</li>
	<li>Insert two separate recording microelectrodes filled with 0.5 M NaCl into the holes using mechanical micromanipulators (at 60&#176;, Figure 1B).</li>
</ol>
2. LFP recordings in bilateral M2 of mice
<ol>
	<li>Lower the left and right glass electrodes slowly into the appropriate coordinates of bilateral M2 (Figure 1C).</li>
	<li>For quality control, test the resistance of each electrode using the differential amplifier before capturing LFPs.</li>
	<li>Set the recording process at 0.1 Hz high-pass and 1,000 Hz low-pass with 1,000x amplification.</li>
	<li>Collect digitized raw LFP data of at least 60 s spontaneous activities in a stable state, with mice breathing evenly at a respiratory rate of 2 breaths per second under anesthesia.</li>
	<li>After recording, slowly raise the electrodes out of the brain, then euthanize the mice by fast cervical dislocation.</li>
	<li>Save the data and analyze it offline.</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>AC/DC Differential Amplifier</td>
			<td>A-M Systems</td>
			<td>Model 3000</td>
			<td></td>
		</tr>
		<tr>
			<td>Analog Digital converter</td>
			<td>Cambridge Electronic Design Ltd.</td>
			<td>Micro1401</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass borosilicate micropipettes</td>
			<td>Nanjing spring teaching experimental equipment company</td>
			<td>161230</td>
			<td>Outer diameter: 1.0mm</td>
		</tr>
		<tr>
			<td>Microelectrode puller</td>
			<td>Narishige</td>
			<td>PC-10</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Guangzhou Chemical Reagent Factory</td>
			<td>7647-14-5</td>
			<td></td>
		</tr>
		<tr>
			<td>Pin microelectrode holder</td>
			<td>World Precision Instruments, INC.</td>
			<td>MEH3SW10</td>
			<td></td>
		</tr>
		<tr>
			<td>Spike2</td>
			<td>Cambridge Electronic Design Ltd.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Stereomicroscope</td>
			<td>Zeiss</td>
			<td>435064-9020-000</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotaxic apparatus</td>
			<td>RWD Life Science</td>
			<td>68045</td>
			<td></td>
		</tr>
		<tr>
			<td>Urethane</td>
			<td>Sigma-Aldrich</td>
			<td>94300</td>
			<td></td>
		</tr>
	</tbody>
</table>

Source: Chen, Y., et al. <a href="https://app.jove.com/v/59310">Evaluation of Hemisphere Lateralization with Bilateral Local Field Potential Recording in Secondary Motor Cortex of Mice.</a> J. Vis. Exp. (2019).
This video demonstrates in vivo recording of local field potential (LFP) from bilateral M2 areas of the mouse brain. An anesthetized mouse is secured and prepared for the procedure. To record LFPs, insert microelectrodes into the target coordinates of the bilateral M2 areas, which are responsible for complex motor movements.

<img alt="Figure 1" src="/files/ftp_upload/22759/22759_figure 1.jpg" /> 
Figure 1: Diagram of the simultaneous LFP recording procedure. (A) Stereotaxic mouse with skull exposed and dura mater removed for in vivo bilateral recording of LFPs in left and right M2. (B) Two glass microelectrodes in touch with the cortical surface in the hole drilled simultaneously. (C) Recording microelectrodes and Ag/AgCl wires as reference electrodes positioned at appropriate sites.

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Animal ...

Begin with an anesthetized mouse with its head secured in a stereotactic frame.
Shave and sterilize the scalp.
Make an incision and remove the residual tissue to expose the skull.
Clean the skull with a disinfectant and drill holes at the target locations.
Remove the outer layer of the brain.
Insert microelectrodes filled with an ionic solution into the holes at a desired angle.
Lower both microelectrodes into the target coordinates of the brain.
Test the electrodes&#39; resistance and adjust the recording settings to ensure the reliable acquisition of local field potential or LFP data.
LFP reflects voltage changes resulting from the collective activity of many neurons within a small region.
Record LFPs from both hemispheres to compare the neural activity between them.

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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.

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

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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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Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in adult mammalian animals remains elusive. In this study, we describe an experimental procedure for measuring calcium dynamics through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy. This method enables recordings and quantifications of neural activity in bilateral mouse brain regions one at a time in the same experiment for a prolonged period in vivo. Key aspects of this method, which can be completed within an hour, include minimally invasive surgery procedures for creating dual optical windows, and the use of 2P imaging. Although we only demonstrate the technique in the S1 area, the method can be applied to other regions of the living brain facilitating the elucidation of structural and functional complexities of brain neural networks.

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

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Non-Invasive Modulation and Robotic Mapping of Motor Cortex in the Developing Brain

Mapping the motor cortex with transcranial magnetic stimulation (TMS) has potential to interrogate motor cortex physiology and plasticity but carries unique challenges in children. Similarly, transcranial direct current stimulation (tDCS) can improve motor learning in adults but has only recently been applied to children. The use of tDCS and emerging techniques like high&#8211;definition tDCS (HD-tDCS) require special methodological considerations in the developing brain. Robotic TMS motor mapping may confer unique advantages for mapping, particularly in the developing brain. Here, we aim to provide a practical, standardized approach for two integrated methods capable of simultaneously exploring motor cortex modulation and motor maps in children. First, we describe a protocol for robotic TMS motor mapping. Individualized, MRI-navigated 12x12 grids centered on the motor cortex guide a robot to administer single-pulse TMS. Mean motor evoked potential (MEP) amplitudes per grid point are used to generate 3D motor maps of individual hand muscles with outcomes including map area, volume, and center of gravity. Tools to measure safety and tolerability of both methods are also included. Second, we describe the application of both tDCS and HD-tDCS to modulate the motor cortex and motor learning. An experimental training paradigm and sample results are described. These methods will advance the application of non-invasive brain stimulation in children.

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Mapping the motor cortex with transcranial magnetic stimulation (TMS) has potential to interrogate motor cortex physiology and plasticity but carries ...

Recording of Local Field Potential from Both Hemispheres of the Mouse Brain

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Recording of Local Field Potential from Both Hemispheres of the Mouse Brain