Neuron–transistor coupling: interpretation of individual extracellular recorded signals

The electrical coupling of randomly migrating neurons from rat explant brain-stem slice cultures to the gates of non-metallized field-effect transistors (FETs) has been investigated. The objective of our work is the precise interpretation of extracellular recorded signal shapes in comparison to the usual patch-clamp protocols to evaluate the possible use of the extracellular recording technique in electrophysiology. The neurons from our explant cultures exhibited strong voltage-gated potassium currents through the plasma membrane. With an improved noise level of the FET set-up, it was possible to record individual extracellular responses without any signal averaging. Cells were attached by patch-clamp pipettes in voltage-clamp mode and stimulated by voltage step pulses. The point contact model, which is the basic model used to describe electrical contact between cell and transistor, has been implemented in the electrical simulation program PSpice. Voltage and current recordings and compensation values from the patch-clamp measurement have been used as input data for the simulation circuit. Extracellular responses were identified as composed of capacitive current and active potassium current inputs into the adhesion region between the cell and transistor gate. We evaluated the extracellular signal shapes by comparing the capacitive and the slower potassium signal amplitudes. Differences in amplitudes were found, which were interpreted in previous work as enhanced conductance of the attached membrane compared to the average value of the cellular membrane. Our results suggest rather that additional effects like electrodiffusion, ion sensitivity of the sensors or more detailed electronic models for the small cleft between the cell and transistor should be included in the coupling model.

[1]  M. Hatten Central nervous system neuronal migration. , 1999, Annual review of neuroscience.

[2]  Andreas Offenhäusser,et al.  Long-term recording system based on field-effect transistor arrays for monitoring electrogenic cells in culture , 1998 .

[3]  A. Hodgkin,et al.  A quantitative description of membrane current and its application to conduction and excitation in nerve , 1990 .

[4]  M. Tacconi Neuronal Death: Is There a Role for Astrocytes? , 1998, Neurochemical Research.

[5]  Peter Fromherz,et al.  FREQUENCY DEPENDENT SIGNAL TRANSFER IN NEURON TRANSISTORS , 1997 .

[6]  A. Offenhäusser,et al.  Electrical recordings from rat cardiac muscle cells using field-effect transistors. , 1999, Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics.

[7]  J. Storm Potassium currents in hippocampal pyramidal cells. , 1990, Progress in brain research.

[8]  H. Aldskogius,et al.  Central neuron–glial and glial–glial interactions following axon injury , 1998, Progress in Neurobiology.

[9]  Stefano Vassanelli,et al.  Transistor Probes Local Potassium Conductances in the Adhesion Region of Cultured Rat Hippocampal Neurons , 1999, The Journal of Neuroscience.

[10]  P. Fromherz,et al.  Noninvasive neuroelectronic interfacing with synaptically connected snail neurons immobilized on a semiconductor chip , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[11]  B. Sakmann,et al.  Single-Channel Recording , 1995, Springer US.

[12]  Peter Fromherz,et al.  Recombinant maxi-K channels on transistor, a prototype of iono-electronic interfacing , 2001, Nature Biotechnology.

[13]  A. Offenhäusser,et al.  Field-effect transistor array for monitoring electrical activity from mammalian neurons in culture. , 1997, Biosensors & bioelectronics.

[14]  B. Sakmann,et al.  Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches , 1981, Pflügers Archiv.

[15]  Weis,et al.  Neuron transistor: Electrical transfer function measured by the patch-clamp technique. , 1993, Physical review letters.

[16]  J. W. Parce,et al.  Detection of cell-affecting agents with a silicon biosensor. , 1989, Science.

[17]  B. Gähwiler Slice cultures of cerebellar, hippocampal and hypothalamic tissue , 1984, Experientia.

[18]  Peter Fromherz,et al.  Extracellular recording with transistors and the distribution of ionic conductances in a cell membrane , 1999, European Biophysics Journal.

[19]  B. Sakmann,et al.  Single-Channel Recording , 1983, Springer US.

[20]  B D DeBusschere,et al.  Portable cell-based biosensor system using integrated CMOS cell-cartridges. , 2001, Biosensors & bioelectronics.

[21]  David W. Tank,et al.  Sealing cultured invertebrate neurons to embedded dish electrodes facilitates long-term stimulation and recording , 1989, Journal of Neuroscience Methods.

[22]  D. Oliver,et al.  Distinct K Currents Result in Physiologically Distinct Cell Types in the Inferior Colliculus of the Rat , 2001, The Journal of Neuroscience.

[23]  F. J. Sigworth,et al.  Design of the EPC-9, a computer-controlled patch-clamp amplifier. 2. Software , 1995, Journal of Neuroscience Methods.

[24]  Martin Jenkner,et al.  BISTABILITV OF MEMBRANE CONDUCTANCE IN CELL ADHESION OBSERVED IN A NEURON TRANSISTOR , 1997 .

[25]  D. Stenger,et al.  Development and Application of Cell-Based Biosensors , 1999, Annals of Biomedical Engineering.

[26]  D. Schmitt-Landsiedel,et al.  A 128 /spl times/ 128 CMOS bio-sensor array for extracellular recording of neural activity , 2003, 2003 IEEE International Solid-State Circuits Conference, 2003. Digest of Technical Papers. ISSCC..

[27]  P. Fromherz,et al.  A neuron-silicon junction: a Retzius cell of the leech on an insulated-gate field-effect transistor. , 1991, Science.

[28]  A. Offenhäusser,et al.  Electrophysiological recordings of patterned rat brain stem slice neurons. , 2002, Biomaterials.

[29]  Andreas Offenhäusser,et al.  Aligned microcontact printing of biomolecules on microelectronic device surfaces , 2001, IEEE Transactions on Biomedical Engineering.

[30]  B. Gähwiler Organotypic monolayer cultures of nervous tissue , 1981, Journal of Neuroscience Methods.

[31]  P. Fromherz,et al.  Neuron–silicon junction with voltage‐gated ionic currents , 1998, The European journal of neuroscience.

[32]  S. Ingebrandt Characterisation of the cell-transistor coupling , 2001 .

[33]  B. Eversmann,et al.  A 128 × 128 CMOS bio-sensor array for extracellular recording of neural activity , 2003 .

[34]  B. Gähwiler,et al.  Distinct modes of channel gating underlie inactivation of somatic K+ current in rat hippocampal pyramidal cells in vitro. , 1996, The Journal of physiology.

[35]  W. Knoll,et al.  Modulation of the growth and guidance of rat brain stem neurons using patterned extracellular matrix proteins , 2001, Neuroscience Letters.

[36]  F. Sigworth Design of the EPC-9, a computer-controlled patch-clamp amplifier. 1. Hardware , 1995, Journal of Neuroscience Methods.

[37]  Joseph J Pancrazio,et al.  A portable microelectrode array recording system incorporating cultured neuronal networks for neurotoxin detection. , 2003, Biosensors & bioelectronics.