Abstract
The aim of this paper was to investigate the influence of four different external factors (acetylcholine, ethanol, temperature and lidocaine hydrochloride) on PC12 quasi-neuronal networks by multielectrode-array-based Voltage Threshold Measurement Method (VTMM). At first, VTMM was employed to measure the lowest amplitude of the voltage stimulating pulses that could just trigger the action potential from PC12 quasi-neuronal networks under normal conditions, and the amplitude was defined as the normal voltage threshold (VTh). Then the changes of the VTh of PC12 quasi-neuronal networks treated by the four external factors were tested respectively. The results showed the normal VTh of PC12 quasi-neuronal networks was 36 mV. The VTh has negative correlation with the concentration of acetylcholine and has positive correlation with the concentration of ethanol. The curves of the correlation of the VTh with temperature and the concentration of lidocaine hydrochloride were U-shaped and Λ-shaped respectively. Comparing with our earlier studies on hippocampal neuronal networks and hippocampal slices, PC12 quasi-neuronal networks not only had the same typical voltage threshold characteristic, but also had similar changes on electrical excitability when treated by the four external factors mentioned above. Therefore, the rapid-formed PC12 quasi-neuronal networks could replace neuronal networks in proper conditions, and VTMM could be used to analyze the influence of external factors on the electrical excitability of PC12 quasi-neuronal networks.
Highlights
Using the voltage threshold principle of voltage clamp during single neuron action potential measurement for reference, Voltage Threshold Measurement Method (VTMM) can quantitatively study the influence of different factors on the electrical excitability of neuronal networks by finding out the voltage threshold (VTh) which could trigger the responses from the networks in resting status and testing the changes of the VTh under the effect of applied factors
The objective of this paper is to investigate the effects of acetylcholine (ACh), ethanol, temperature and lidocaine hydrochloride (LDH) on VTh of PC12 quasi-neuronal networks through VTMM, respectively, and further to compare the results with that of our previous studies completed with hippocampal neuronal networks and hippocampal slices
According to the principle of VTMM and the definition of normal VTh, the influence of the applied external factors was inhibitory to PC12 quasi-neuronal networks when the VTh was higher than the normal VTh and was excitatory when the VTh was lower than the normal VTh
Summary
Thomas Jr. invented microelectrode array (MEA), which could be used to measure the electrical activities of neuronal networks cultured on its surface and implemented patch clamp’s limitation that the electrophysiological characteristics of only one single neuron could be tested each time [1]. In order to study the electrophysiological characteristics of neuronal networks in an easier and faster way, our group has developed the Voltage Threshold Measurement Method (VTMM) based on MEA techniques [6]. Using the voltage threshold principle of voltage clamp during single neuron action potential measurement for reference, VTMM can quantitatively study the influence of different factors on the electrical excitability of neuronal networks by finding out the voltage threshold (VTh) which could trigger the responses from the networks in resting status and testing the changes of the VTh under the effect of applied factors. It is valuable to find a simple network model as a substitute for normal neuronal networks for VTMM and studying the electrophysiological characteristics of neuronal networks in an easier and faster way
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