Dissociated cortical networks show spontaneously correlated activity patterns during in vitro development

In vitro cultured neuronal networks coupled to microelectrode arrays (MEAs) constitute a valuable experimental model for studying changes in the neuronal dynamics at different stages of development. After a few days in culture, neurons start to connect each other with functionally active synapses, forming a random network and displaying spontaneous electrophysiological activity. The patterns of collective rhythmic activity change in time spontaneously during in vitro development. Such activity-dependent modifications play a key role in the maturation of the network and reflect changes in the synaptic efficacy, fact widely recognized as a cellular basis of learning, memory and developmental plasticity. Getting advantage from the possibilities offered by the MEAs, the aim of our study is to analyze and characterize the natural changes in dynamics of the electrophysiological activity at different ages of the culture, identifying peculiar steps of the spontaneous evolution of the network. The main finding is that between the second and the third week of culture, the network completely changes its electrophysiological patterns, both in terms of spiking and bursting activity and in terms of cross-correlation between pairs of active channels. Then the maturation process can be characterized by two main phases: modulation and shaping in the synaptic functional connectivity of the network (within the first and second week) and general moderate correlated activity, spread over the entire network, with connections properly formed and stabilized (within the fourth and fifth week).

[1]  P. S. Wolters,et al.  Longterm stability and developmental changes in spontaneous network burst firing patterns in dissociated rat cerebral cortex cell cultures on multielectrode arrays , 2004, Neuroscience Letters.

[2]  William Bialek,et al.  Spikes: Exploring the Neural Code , 1996 .

[3]  G. Gross,et al.  NMDA receptor-dependent periodic oscillations in cultured spinal cord networks. , 2001, Journal of neurophysiology.

[4]  M. Grattarola,et al.  In vitro 2-D networks of neurons characterized by processing the signals recorded with a planar microtransducer array , 1997, IEEE Transactions on Biomedical Engineering.

[5]  B. Sutor,et al.  Dye coupling between pyramidal neurons in developing rat prefrontal and frontal cortex is reduced by protein kinase A activation and dopamine , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[6]  M. Nedergaard,et al.  Direct signaling from astrocytes to neurons in cultures of mammalian brain cells. , 1994, Science.

[7]  Eisaku Maeda,et al.  Experimental analysis of neuronal dynamics in cultured cortical networks and transitions between different patterns of activity , 1997, Biological Cybernetics.

[8]  Gregory J Brewer,et al.  Isolation and culture of adult rat hippocampal neurons , 1997, Journal of Neuroscience Methods.

[9]  H. Robinson,et al.  Spontaneous periodic synchronized bursting during formation of mature patterns of connections in cortical cultures , 1996, Neuroscience Letters.

[10]  Daniel A. Wagenaar,et al.  The Neurally Controlled Animat: Biological Brains Acting with Simulated Bodies , 2001, Auton. Robots.

[11]  Thomas M. McKenna,et al.  Enabling Technologies for Cultured Neural Networks , 1994 .

[12]  M. Corner,et al.  Dynamics and plasticity in developing neuronal networks in vitro. , 2005, Progress in brain research.

[13]  R. Balice-Gordon,et al.  Astrocytes Regulate Inhibitory Synapse Formation via Trk-Mediated Modulation of Postsynaptic GABAA Receptors , 2005, The Journal of Neuroscience.

[14]  Thoralf Opitz,et al.  Spontaneous development of synchronous oscillatory activity during maturation of cortical networks in vitro. , 2002, Journal of neurophysiology.

[15]  L. Martin,et al.  Long-term culture of mouse cortical neurons as a model for neuronal development, aging, and death. , 2002, Journal of neurobiology.

[16]  G. Gross,et al.  A closed flow chamber for long-term multichannel recording and optical monitoring , 1994, Journal of Neuroscience Methods.

[17]  Masahiro Kawahara,et al.  Formation and maturation of synapses in primary cultures of rat cerebral cortical cells: an electron microscopic study , 1993, Neuroscience Research.

[18]  G. P. Moore,et al.  Neuronal spike trains and stochastic point processes. I. The single spike train. , 1967, Biophysical journal.

[19]  Shimon Marom,et al.  Learning in ex-vivo developing networks of cortical neurons. , 2005, Progress in brain research.

[20]  H. Robinson,et al.  The mechanisms of generation and propagation of synchronized bursting in developing networks of cortical neurons , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[21]  C. Shatz,et al.  Synaptic Activity and the Construction of Cortical Circuits , 1996, Science.

[22]  Y. Ben-Ari.  Developing networks play a similar melody , 2001, Trends in Neurosciences.

[23]  Shimon Marom,et al.  Development, learning and memory in large random networks of cortical neurons: lessons beyond anatomy , 2002, Quarterly Reviews of Biophysics.

[24]  Steve M. Potter,et al.  Controlling Bursting in Cortical Cultures with Closed-Loop Multi-Electrode Stimulation , 2005, The Journal of Neuroscience.

[25]  F. Pfrieger,et al.  Synaptic efficacy enhanced by glial cells in vitro. , 1997, Science.

[26]  J. Palva,et al.  Synaptic GABA(A) activation inhibits AMPA-kainate receptor-mediated bursting in the newborn (P0-P2) rat hippocampus. , 2000, Journal of neurophysiology.

[27]  M. Heuschkel,et al.  The generation of rhythmic activity in dissociated cultures of rat spinal cord , 2001, The European journal of neuroscience.

[28]  P. Renaud,et al.  Spatiotemporal characterization of rhythmic activity in rat spinal cord slice cultures , 2001, The European journal of neuroscience.

[29]  J. Matias Palva,et al.  Fast Network Oscillations in the Newborn Rat HippocampusIn Vitro , 2000, The Journal of Neuroscience.

[30]  C. Goodman,et al.  Mechanisms and molecules that control growth cone guidance. , 1996, Annual review of neuroscience.

[31]  T. Sejnowski,et al.  Correlated neuronal activity and the flow of neural information , 2001, Nature Reviews Neuroscience.

[32]  Luca Berdondini,et al.  Towards an embodied in vitro electrophysiology: the NeuroBIT project , 2004, Neurocomputing.

[33]  K. Muramoto,et al.  Frequency of synchronous oscillations of neuronal activity increases during development and is correlated to the number of synapses in cultured cortical neuron networks , 1993, Neuroscience Letters.

[34]  Michael J. O'Donovan,et al.  Blockade and Recovery of Spontaneous Rhythmic Activity after Application of Neurotransmitter Antagonists to Spinal Networks of the Chick Embryo , 1998, The Journal of Neuroscience.

[35]  G Shahaf,et al.  Learning in Networks of Cortical Neurons , 2001, The Journal of Neuroscience.

[36]  A. Peinado.  Immature neocortical neurons exist as extensive syncitial networks linked by dendrodendritic electrical connections. , 2001, Journal of neurophysiology.

[37]  H. Müller,et al.  GABA and glutamate receptor development of cultured neurons from rat hippocampus, septal region, and neocortex , 1990, Synapse.

[38]  Michael J. O'Donovan.  The origin of spontaneous activity in developing networks of the vertebrate nervous system , 1999, Current Opinion in Neurobiology.

[39]  Wim L. C. Rutten,et al.  Long-term characterization of firing dynamics of spontaneous bursts in cultured neural networks , 2004, IEEE Transactions on Biomedical Engineering.

[40]  N. Ziv,et al.  Dopamine-induced dispersion of correlations between action potentials in networks of cortical neurons. , 2004, Journal of neurophysiology.

[41]  G. Brewer,et al.  Optimized survival of hippocampal neurons in B27‐supplemented neurobasal™, a new serum‐free medium combination , 1993, Journal of neuroscience research.

[42]  Steve M. Potter,et al.  A new approach to neural cell culture for long-term studies , 2001, Journal of Neuroscience Methods.

[43]  V. Parpura,et al.  Vesicular Glutamate Transporter-Dependent Glutamate Release from Astrocytes , 2004, Journal of Neuroscience.

[44]  Vincent Torre,et al.  Toward the neurocomputer: image Processing and pattern recognition with neuronal cultures , 2005, IEEE Transactions on Biomedical Engineering.

[45]  Alessandro Vato,et al.  Spike Manager: A New Tool for Spontaneous and Evoked Neuronal Networks Activity Characterization , 2003, Neurocomputing.

[46]  H. Lüscher,et al.  Emerging Network Activity in Dissociated Cultures of Neocortex: Novel Electrophysiological Protocols and Mathematical Modeling , 2006 .

[47]  Giorgio Carmignoto,et al.  Reciprocal communication systems between astrocytes and neurones , 2000, Progress in Neurobiology.

[48]  A Kawana,et al.  Periodic synchronized bursting and intracellular calcium transients elicited by low magnesium in cultured cortical neurons. , 1993, Journal of neurophysiology.

[49]  A. Araque,et al.  Tripartite synapses: glia, the unacknowledged partner , 1999, Trends in Neurosciences.

[50]  M Giugliano,et al.  Single-neuron discharge properties and network activity in dissociated cultures of neocortex. , 2004, Journal of neurophysiology.

[51]  S. Nelson,et al.  Homeostatic plasticity in the developing nervous system , 2004, Nature Reviews Neuroscience.

[52]  R. Yuste,et al.  Neuronal domains in developing neocortex. , 1992, Science.

[53]  M Tedesco,et al.  In vitro cortical neuronal networks as a new high-sensitive system for biosensing applications. , 2005, Biosensors & bioelectronics.

[54]  Vincent Torre,et al.  Statistical properties of information processing in neuronal networks , 2005, The European journal of neuroscience.

[55]  M. Feller,et al.  Spontaneous Correlated Activity in Developing Neural Circuits , 1999, Neuron.

[56]  P G Nelson,et al.  Activity-dependent development of the vertebrate nervous system. , 1992, International review of neurobiology.

[57]  M. Chiappalone,et al.  Networks of neurons coupled to microelectrode arrays: a neuronal sensory system for pharmacological applications. , 2003, Biosensors & bioelectronics.

[58]  Sergio Martinoia,et al.  Electrophysiological activity modulation by chemical stimulation in networks of cortical neurons coupled to microelectrode arrays : A biosensor for neuropharmacological applications , 2005 .

[59]  Joseph J Pancrazio,et al.  Measuring synchronization in neuronal networks for biosensor applications. , 2004, Biosensors & bioelectronics.

[60]  G. Turrigiano.  Homeostatic plasticity in neuronal networks: the more things change, the more they stay the same , 1999, Trends in Neurosciences.

[61]  Lawrence C. Katz,et al.  Coordinate activity in retinal and cortical development , 1993, Current Opinion in Neurobiology.

[62]  H. Robinson,et al.  Simultaneous induction of pathway-specific potentiation and depression in networks of cortical neurons. , 1999, Biophysical journal.

[63]  C. Knox,et al.  Detection of neuronal interactions using correlation analysis , 1981, Trends in Neurosciences.

[64]  Yen-Chung Chang,et al.  Development of excitatory synapses in cultured neurons dissociated from the cortices of rat embryos and rat pups at birth , 2002, Journal of neuroscience research.

[65]  Alessandro Vato,et al.  Burst detection algorithms for the analysis of spatio-temporal patterns in cortical networks of neurons , 2005, Neurocomputing.

[66]  Yasuhiko Jimbo,et al.  Activity-dependent enhancement in the reliability of correlated spike timings in cultured cortical neurons , 1999, Biological Cybernetics.