Simultaneous recording of the action potential and its whole-cell associated ion current on NG108-15 cells cultured over a MWCNT electrode

Abstract

It is well known that, in excitable cells, the dynamics of the ion currents (I i) is extremely important to determine both the magnitude and time course of an action potential (A p). To observe these two processes simultaneously, we cultured NG108-15 cells over a multi-walled carbon nanotubes electrode (MWCNTe) surface and arranged a two independent Patch Clamp system configuration (Bi-Patch Clamp). The first system was used in the voltage or current clamp mode, using a glass micropipette as an electrode. The second system was modified to connect the MWCNTe to virtual ground. While the A p was recorded through the micropipette electrode, the MWCNTe was used to measure the underlying whole-cell current. This configuration allowed us to record both the membrane voltage (V m) and the current changes simultaneously. Images acquired by atomic force microscopy (AFM) and scanning electron microscopy (SEM) indicate that cultured cells developed a complex network of neurites, which served to establish the necessary close contact and strong adhesion to the MWCNTe surface. These features were a key factor to obtain the recording of the whole-cell I i with a high signal to noise ratio (SNR). The experimental results were satisfactorily reproduced by a theoretical model developed to simulate the proposed system. Besides the contribution to a better understanding of the fundamental mechanisms involved in cell communication, the developed method could be useful in cell physiology studies, pharmacology and diseases diagnosis.

Carbon-Nanotube-Wrapped Spider Silks for Directed Cardiomyocyte Growth and Electrophysiological Detection

“…The current traces of devices D1–D3 demonstrated regular firing frequencies (Figure d, left panels) and signal shapes (Figure d, right panels). The electrophysiological signal observed is similar to that reported in other studies of the whole–cell ion current recorded by the multiwalled CNT electrode . Meanwhile, the stable and regular current traces further suggested that robust interfaces were formed between CNT–silk and cardiomycytes.…”

“…The electrophysiological signal observed is similar to that reported in other studies of the whole−cell ion current recorded by the multiwalled CNT electrode. 22 Meanwhile, the stable and regular current traces further suggested that robust interfaces were formed between CNT−silk and cardiomycytes. These results show that the CNT−silk-based devices are capable of recording the spontaneous electrophysiological activity of different cardiomy- cytes simultaneously, and the electrophysiological changes that might occur during the primary culture can be identified and characterized.…”

Simultaneous recording of electrical activity and the underlying ionic currents in NG108-15 cells cultured on gold substrate

“…The latter problem had been partially solved by other approaches ( Dietrich et al., 2002 ; Banyasz et al., 2011 ; Barra, 1996 ; Bean, 2007 ; Berecki et al., 2005 ; Doerr et al., 1990 ; Economo et al., 2010 ; Nowotny et al., 2006 ; Molnar and Hickman, 2007 ; Wilders, 2006 ; Hille, 1994 ; Levitan, 1994 ). Very recently, it was fully solved by using carbon nanotubes (maintained at virtual ground) as a substrate for cell culturing ( Morales-Reyes et al., 2016 ).…”

“…with good signal to noise ratio) of the action potentials and the underlying ionic currents ( Dietrich et al., 2002 ; Banyasz et al., 2011 ; Barra, 1996 ; Bean, 2007 ; Berecki et al., 2005 ; Doerr et al., 1990 ; Economo et al., 2010 ; Nowotny et al., 2006 ; Molnar and Hickman, 2007 ; Wilders, 2006 ). Recently, this limitation was overcome by using carbon nanotubes as a substrate for cell culturing ( Morales-Reyes et al., 2016 ). In this work, we propose that gold substrates can be used instead of carbon nanotubes to perform simultaneous measurements of electrical activity and the underlying membrane currents.…”

Functional Consequences of Keratan Sulfate Sulfation in Electrosensory Tissues and in Neuronal Regulation

“…The culture of neurons on peptide coated gold surfaces [265,266] and multielectrode microarrays has facilitated the measurement of generated membrane action potentials and whole-cell ion currents. [267][268][269] Culture of NG108-15 cells on gold surfaces has improved the analysis of intracellular ioncurrents and membrane potentials in cultured neurons. [98] AFM and SEM has also been applied to the characterization of these cells [270][271][272] and to determine the roles of regulatory ion channels, [273,274] and how these are modulated by G-coupled receptors [275] and the sensory input of fillopodia in neuronal cultures.…”