Development of multi-electrode array screening for anticonvulsants in acute rat brain slices.
The acute hippocampal brain slice preparation is an important in vitro screening tool for potential anticonvulsants. Application of 4-aminopyridine (4-AP) or removal of external Mg(2+) ions induces epileptiform bursting in slices which is analogous to electrical brain activity seen in status epilepticus states. We have developed these epileptiform models for use with multi-electrode arrays (MEAs), allowing recording across the hippocampal slice surface from 59 points. We present validation of this novel approach and analyses using two anticonvulsants, felbamate and phenobarbital, the effects of which have already been assessed in these models using conventional extracellular recordings. In addition to assessing drug effects on commonly described parameters (duration, amplitude and frequency), we describe novel methods using the MEA to assess burst propagation speeds and the underlying frequencies that contribute to the epileptiform activity seen. Contour plots are also used as a method of illustrating burst activity. Finally, we describe hitherto unreported properties of epileptiform bursting induced by 100 microM 4-AP or removal of external Mg(2+) ions. Specifically, we observed decreases over time in burst amplitude and increase over time in burst frequency in the absence of additional pharmacological interventions. These MEA methods enhance the depth, quality and range of data that can be derived from the hippocampal slice preparation compared to conventional extracellular recordings. It may also uncover additional modes of action that contribute to anti-epileptiform drug effects.
Hill
Andrew J
AJ
Reading School of Pharmacy, University of Reading, Whiteknights, Reading RG6 6UB, United Kingdom. a.j.hill@reading.ac.uk
Jones
Nicholas A
NA
Williams
Claire M
CM
Stephens
Gary J
GJ
Whalley
Benjamin J
BJ
D015761:4-Aminopyridine / Q000009:adverse effects; D000818:Animals; D000831:Animals, Newborn; D000927:Anticonvulsants / Q000494:pharmacology*; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation* / Q000379:methods; D004566:Electrodes; D004594:Electrophysiology / Q000379:methods; D004827:Epilepsy / Q000188:drug therapy / Q000209:etiology; D000078328:Felbamate; D005260:Female; D006624:Hippocampus / Q000187:drug effects; D066298:In Vitro Techniques; D008275:Magnesium Deficiency / Q000150:complications; D008564:Membrane Potentials / Q000187:drug effects; D010634:Phenobarbital / Q000494:pharmacology; D048448:Phenylcarbamates / Q000494:pharmacology; D011409:Propylene Glycols / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors
D000927:Anticonvulsants; D048448:Phenylcarbamates; D011409:Propylene Glycols; D015761:4-Aminopyridine; D000078328:Felbamate; D010634:Phenobarbital