Functional screening of traditional antidepressants with primary cortical neuronal networks grown on multielectrode neurochips

We optimized the novel technique of multielectrode neurochip recordings for the rapid and efficient screening of neuroactivity. Changes in the spontaneous activity of cultured networks of primary cortical neurons were quantified to evaluate the action of drugs on the firing dynamics of complex network activity. The multiparametric assessment of electrical activity changes caused by psychoactive herbal extracts from Hypericum, Passiflora and Valeriana, and various combinations thereof revealed a receptor‐specific and concentration‐dependent inhibition of the firing patterns. The spike and burst rates showed significant substance‐dependent effects and significant differences in potency. The effects of specific receptor blockades on the inhibitory responses provided evidence that the herbal extracts act on gamma‐amino butyric acid (GABA) and serotonin (5‐HT) receptors, which are recognized targets of pharmacological antidepressant treatment. A biphasic effect, serotonergic stimulation of activity at low concentrations that is overridden by GABAergic inhibition at higher concentrations, is apparent with Hypericum alone and the triple combination of the extracts. The more potent neuroactivity of the triple combination compared to Hypericum alone and the additive effect of Passiflora and Valeriana suggest a synergy between constituent herbal extracts. The extracts and their combinations affected the set of derived activity parameters in a concomitant manner suggesting that all three constituent extracts and their combinations have largely similar modes of action. This study also demonstrates the sensitivity, selectivity and robustness of neurochip recordings for high content screening of complex mixtures of neuroactive substances and for providing multiparametric information on neuronal activity changes to assess the therapeutic potential of psychoactive substances.

[1]  M. Mahoney,et al.  St. John's wort. , 2005, American family physician.

[2]  H. Schulz,et al.  Investigation of Pharmacokinetic Data of Hypericin, Pseudohypericin, Hyperforin and the Flavonoids Quercetin and Isorhamnetin Revealed from Single and Multiple Oral Dose Studies with a Hypericum Extract Containing Tablet in Healthy Male Volunteers , 2005, Arzneimittelforschung.

[3]  A. Nahrstedt,et al.  Biologically Active and Other Chemical Constituents of the Herb of Hypericum perforatum L. , 1997, Pharmacopsychiatry.

[4]  J. Cott.  In Vitro Receptor Binding and Enzyme Inhibition by Hypericum perforatum Extract , 1997, Pharmacopsychiatry.

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

[6]  Effect of blood plasma components from patients with hepatic encephalopathy on electrophysiological activity of primary frontal cortex networks in vitro , 2003 .

[7]  Pal Pacher,et al.  Trends in the development of new antidepressants. Is there a light at the end of the tunnel? , 2004, Current medicinal chemistry.

[8]  P. Connolly,et al.  Advantages of using microfabricated extracellular electrodes for in vitro neuronal recording , 1995, Journal of neuroscience research.

[9]  S. Hansen,et al.  Adhyperforin as a contributor to the effect of Hypericum perforatum L. in biochemical models of antidepressant activity. , 2001, Life sciences.

[10]  W. Müller,et al.  Inhibition of Synaptosomal Uptake of 3H-L-glutamate and 3H-GABA by Hyperforin, a Major Constituent of St. John's Wort: The Role of Amiloride Sensitive Sodium Conductive Pathways , 2000, Neuropsychopharmacology.

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

[12]  Veronika Butterweck,et al.  Mechanism of Action of St John’s Wort in Depression , 2003, CNS drugs.

[13]  G. Gross,et al.  Drug evaluations using neuronal networks cultured on microelectrode arrays. , 2000, Biosensors & bioelectronics.

[14]  C. Cavadas,et al.  In vitro study on the interaction of Valeriana officinalis L. extracts and their amino acids on GABAA receptor in rat brain. , 1995, Arzneimittel-Forschung.

[15]  P. Schmidt,et al.  Quantitative analysis of the active components and the by-products of eight dry extracts of Hypericum perforatum L. (St John's Wort). , 2002, Phytochemical analysis : PCA.

[16]  R. Chamuleau,et al.  Encephalopathy and Nitrogen Metabolism in Liver Failure , 2003, Springer Netherlands.

[17]  S. Caccia.  Antidepressant-like components of Hypericum perforatum extracts: an overview of their pharmacokinetics and metabolism. , 2005, Current drug metabolism.

[18]  B. Gaster,et al.  St John's wort for depression: a systematic review. , 2000, Archives of internal medicine.

[19]  F. Petty.  GABA and mood disorders: a brief review and hypothesis. , 1995, Journal of affective disorders.

[20]  E. Ernst,et al.  St John's wort: Prozac from the plant kingdom. , 2001, Trends in pharmacological sciences.

[21]  G. Gross,et al.  Quantification of acute neurotoxic effects of trimethyltin using neuronal networks cultured on microelectrode arrays. , 2000, Neurotoxicology.

[22]  P Pacher,et al.  Current trends in the development of new antidepressants. , 2001, Current medicinal chemistry.

[23]  P G Nelson,et al.  Mouse spinal cord in cell culture. I. Morphology and intrinsic neuronal electrophysiologic properties. , 1977, Journal of neurophysiology.

[24]  R. Soulimani,et al.  Behavioural effects of Passiflora incarnata L. and its indole alkaloid and flavonoid derivatives and maltol in the mouse. , 1997, Journal of ethnopharmacology.

[25]  C. Faro,et al.  Pharmacological effects of extracts from Valeriana adscendens Trel. II. Effects on GABA uptake and amino acids , 2003, Phytotherapy research : PTR.

[26]  E. Perl,et al.  Microelectrode arrays for stimulation of neural slice preparations , 1997, Journal of Neuroscience Methods.

[27]  J. Massagué.  TGF-beta signal transduction. , 1998, Annual review of biochemistry.

[28]  C. Legéndy,et al.  Bursts and recurrences of bursts in the spike trains of spontaneously active striate cortex neurons. , 1985, Journal of neurophysiology.

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

[30]  M. Claeys,et al.  Mass spectral characterization of C-glycosidic flavonoids isolated from a medicinal plant (Passiflora incarnata). , 1991, Journal of chromatography.

[31]  W. Burkard,et al.  Extracts and constituents of Hypericum perforatum inhibit the binding of various ligands to recombinant receptors expressed with the Semliki Forest virus system. , 1999, Journal of receptor and signal transduction research.

[32]  Guenter W. Gross,et al.  Recording of spontaneous activity with photoetched microelectrode surfaces from mouse spinal neurons in culture , 1982, Journal of Neuroscience Methods.

[33]  G. Gross,et al.  Transparent indium-tin oxide electrode patterns for extracellular, multisite recording in neuronal cultures , 1985, Journal of Neuroscience Methods.

[34]  S. Kumar,et al.  Anti-anxiety studies on extracts of Passiflora incarnata Linneaus. , 2001, Journal of ethnopharmacology.

[35]  Josef Hölzl,et al.  Lignans isolated from valerian: identification and characterization of a new olivil derivative with partial agonistic activity at A(1) adenosine receptors. , 2002, Journal of natural products.

[36]  G. Gross,et al.  Stimulation of monolayer networks in culture through thin-film indium-tin oxide recording electrodes , 1993, Journal of Neuroscience Methods.

[37]  J. Pine.  Recording action potentials from cultured neurons with extracellular microcircuit electrodes , 1980, Journal of Neuroscience Methods.

[38]  A. Hirshfeld,et al.  Inhibition of vesicular uptake of monoamines by hyperforin. , 2002, Life sciences.

[39]  M. Baraldi,et al.  Behavioral characterisation of the flavonoids apigenin and chrysin. , 2000, Fitoterapia.

[40]  J. Lisman.  Bursts as a unit of neural information: making unreliable synapses reliable , 1997, Trends in Neurosciences.

[41]  R. Baughman,et al.  Primary culture of identified neurons from the visual cortex of postnatal rats , 1986, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[42]  B. Roth,et al.  In vitro receptor screening of pure constituents of St. John's wort reveals novel interactions with a number of GPCRs , 2002, Psychopharmacology.

[43]  Chun-Su Yuan,et al.  The Gamma-Aminobutyric Acidergic Effects of Valerian and Valerenic Acid on Rat Brainstem Neuronal Activity , 2004, Anesthesia and analgesia.

[44]  G. Gross,et al.  A new fixed-array multi-microelectrode system designed for long-term monitoring of extracellular single unit neuronal activity in vitro , 1977, Neuroscience Letters.

[45]  S. Bhattacharya,et al.  Hyperforin as a possible antidepressant component of hypericum extracts. , 1998, Life sciences.

[46]  Bruce C. Wheeler,et al.  Gold-coated microelectrode array with thiol linked self-assembled monolayers for engineering neuronal cultures , 2004, IEEE Transactions on Biomedical Engineering.

[47]  V. Schulz.  [Incidence and clinical relevance of interactions and side-effects of hypericum preparations]. , 2000, Praxis.

[48]  M Bove,et al.  An array of Pt-tip microelectrodes for extracellular monitoring of activity of brain slices. , 1999, Biosensors & bioelectronics.

[49]  Makoto Taketani,et al.  Chronic multichannel recordings from organotypic hippocampal slice cultures: protection from excitotoxic effects of NMDA by non-competitive NMDA antagonists , 2002, Journal of Neuroscience Methods.

[50]  Steve M. Potter,et al.  Distributed Processing in Cultured Neuronal Networks Chapter 4 , 2001 .

[51]  J. Ortiz,et al.  Effects of Valeriana Officinalis Extracts on [3H]Flunitrazepam Binding, Synaptosomal [3H]GABA Uptake, and Hippocampal [3H]GABA Release , 1999, Neurochemical Research.

[52]  H. Volz,et al.  Potential treatment for subthreshold and mild depression: a comparison of St. John's wort extracts and fluoxetine. , 2000, Comprehensive psychiatry.

[53]  J. Streit,et al.  Regular oscillations of synaptic activity in spinal networks in vitro. , 1993, Journal of neurophysiology.

[54]  Guenter W Gross,et al.  Histiotypic electrophysiological responses of cultured neuronal networks to ethanol. , 2003, Alcohol.

[55]  Erik D Herzog,et al.  Circadian rhythms in mouse suprachiasmatic nucleus explants on multimicroelectrode plates , 1997, Brain Research.

[56]  Jürg Streit,et al.  Mechanisms controlling bursting activity induced by disinhibition in spinal cord networks , 2002, The European journal of neuroscience.

[57]  L. Ballerini,et al.  Spontaneous rhythmic bursts induced by pharmacological block of inhibition in lumbar motoneurons of the neonatal rat spinal cord. , 1996, Journal of neurophysiology.

[58]  Anupam Sharma,et al.  Passiflora: a review update. , 2004, Journal of ethnopharmacology.

[59]  Guenter W. Gross,et al.  Potassium and calcium channel dependence of bursting in cultured neuronal networks , 1994, Brain Research.

[60]  P J Houghton,et al.  The Scientific Basis for the Reputed Activity of Valerian , 1998, The Journal of pharmacy and pharmacology.

[61]  G. Gross,et al.  Substance identification by quantitative characterization of oscillatory activity in murine spinal cord networks on microelectrode arrays , 2004, The European journal of neuroscience.

[62]  M. Schubert-Zsilavecz,et al.  Hyperforin modifies neuronal membrane properties in vivo , 2004, Neuroscience Letters.

[63]  J. Medina,et al.  Possible anxiolytic effects of chrysin, a central benzodiazepine receptor ligand isolated from Passiflora Coerulea , 1994, Pharmacology Biochemistry and Behavior.