assessing the effects of toxins on chick embryo neural network development using multielectrode arrays

Assessing the Effects of Toxins on Chick Embryo Neural Network Development Using Multielectrode Arrays

On the day of acquisition, replace the culture medium with fresh neurobasal medium, and return the arrays to the CO2 incubator for a minimum of two hours before recording. Start the recording software and set the temperature of the multi-electrode array to 37 degrees Celsius by clicking on the temperature icon. Set the acquisition parameters by selecting streams in the left window panel, and right-clicking on the muse icon. Then, select Add Processing and Spike Detector and click OK in the pop-up window. Spike Detector 6 by STD will appear below the file name.
Next, right-click on the Spike Detector, select Add Processing and Burst Detector, and click OK in the pop-up window. Burst Detector ISI will appear below the muse icon. Then right-click on Burst Detector, select Add Processing and Neural Statistics Compiler. In the pop-up window, ensure that File Header, Aggregated Well Statistics, and Synchrony are selected. Click OK.
Statistics Compiler will appear below. Click on the clock icon at the bottom of the screen and change the information in the Settings section to record every 5.1 minutes and record for five minutes. This will be started immediately and will be executed once. After retrieving the multi-electrode array from the incubator, place it on the recording unit and lock it into place. Start recording the network activity by clicking on Start Record in the scheduled recording section.

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1. Recording Neuronal Network Activity
<ol>
	<li>On the day of acquisition, exchange the culture medium with fresh neurobasal medium and return the MEAs to the CO2 incubator for a minimum of 2 h before recording. Start the recording software and set the temperature of the MEA to 37 &#176;C by clicking on the temperature icon. 
	NOTE: Recording can begin as early as day 3 of plating and can continue as long as the cultures are alive.</li>
	<li>Set up the acquisition parameters by right-clicking on the &#34;Muse&#34; icon (under Streams in the left window panel) and select &#34;Add Processing&#34; and &#34;Spike Detector&#34; and click OK in the pop up window. &#34;Spike Detector (6 x STD)&#34; will appear below the file name.</li>
	<li>Next right click on the Spike Detector, select &#34;Add Processing&#34; and &#34;Burst Detector&#34; and click OK in the pop up window. &#34;Burst Detector&#34; (ISI) will appear below the Muse icon.</li>
	<li>Next right click on Burst Detector, select &#34;Add Processing&#34; and &#34;Neural Statistics Compiler.&#34; In the pop up window, ensure that File Header, Aggregated Well Statistics and Synchrony are selected. Click OK. &#34;Statistics Compiler&#34; will appear below. 
	NOTE: This will set the adaptive threshold crossing (6x STD filter), the inter-spike interval at 100 ms with a minimum number of spikes at 5, the mean firing rate detection at 10 s with a synchrony parameter at 20 ms, and the minimum spike rate at 0.083333 spikes per min.</li>
	<li>Set up the scheduled recording for the recording time of 5 minutes (300,000 ms) by clicking on the clock icon at the bottom. This is achieved by changing the information in the setting section to record every 5.1 minutes and record for 5 minutes. This will be started immediately and will be executed once.</li>
	<li>Ensure that the temperature of the MEA has reached 37 &#176;C before moving the MEA. Remove the MEA from the incubator, place it on the recording unit, and lock down the MEA. Start recording the network activity by clicking on the start record in the scheduled recording section or the record icon in the &#34;File Play&#34; window. Typically, a recording time of 5 minutes (300,000 ms) is sufficient, although longer recordings of up to 10 minutes can be obtained.</li>
	<li>After recording, return the MEA to the incubator. 
	NOTE: Care must be taken to maintain sterility and to minimize the time the MEA is outside the incubator.</li>
</ol>

Source: Sanchez, K. R., et.al. <a href="https://app.jove.com/v/56300">Assessment of the Effects of Endocrine Disrupting Compounds on the Development of Vertebrate Neural Network Function Using Multi-electrode Arrays.</a> J. Vis. Exp. (2018).
This video demonstrates the use of multi-electrode arrays (MEA) to study the effects of toxins on early embryonic chick neuronal cultures. By monitoring the synchronous activity of the neurons in the neuronal network, the MEA captures the functional dynamics of the neuronal network in response to toxin exposure. Reduced synchrony and firing rates upon toxin exposure highlight its detrimental impact on neuronal network maturation and function.

1. Recording Neuronal Network Activity

	On the day of acquisition, exchange the culture medium with fresh neurobasal medium and return the MEAs to the ...

Begin with adherent cultures of early embryonic chick neurons on extracellular matrix-coated multielectrode arrays, or MEAs.
The base of the MEAs contains grids of tiny electrodes that record neuronal activity.
In the test culture, neurons grow in the presence of a toxin, while in the control culture, neurons grow without the toxin.
Before recording, replace the media in both MEAs with toxin-free media and incubate briefly.
Next, place one MEA in the recording device and begin recording.
Neurons communicate via electrical impulses called action potentials or spikes.
At the synapse, these action potentials trigger neurotransmitter release, propagating the signal to the next neuron.
The MEA electrodes record the spikes generated by the synaptically connected neurons.&#160;
In the control condition, the healthy mature neuronal network exhibits synchronous spiking activity.
In the test culture, reduced spiking and synchrony suggest impaired neuronal network maturation caused by the toxin.

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Assessing the Effects of Toxins on Chick Embryo Neural Network Development Using Multielectrode Arrays

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