Organotypic Cultures as Tools for Testing Neuroactive Drugs – Link Between In-Vitro and In-Vivo Experiments

B.      Drexler; H.      Hentschke; B.      Antkowiak; C.      Grasshoff

Abstract

The development of neuroactive drugs is a time consuming procedure. Candidate drugs must be run through a battery of tests, including receptor studies and behavioural tests on animals. As a rule, numerous substances with promising properties as assessed in receptor studies must be eliminated from the development pipeline in advanced test phases because of unforeseen problems like intolerable side-effects or unsatisfactory performance in the whole organism. Clearly, test systems of intermediate complexity would alleviate this inefficiency. In this review, we propose cultured organotypic brain slices as model systems that could bridge the ‘interpolation gap’ between receptors and the brain, with a focus on the development of new general anaesthetics with lesser side effects. General anaesthesia is based on the modulation of neurotransmitter receptors and other conductances located on neurons in diverse brain regions, including cerebral cortex and spinal cord. It is well known that different components of general anaesthesia, e.g. hypnosis and immobility, are produced by the depression of neuronal activity in distinct brain regions. The ventral horn of the spinal cord is an important structure for the induction of immobility. Thus, the potentially immobilizing effects of a newly designed drug can be estimated from its depressant effect on neuronal network activity in cultured spinal slices. A drugs sedative and hypnotic potential can be examined in cortical cultures. Combined with genetically engineered mice, this approach can point to receptor subtypes most relevant to the drugs intended net effect and in return can help in the design of more selective drugs. In conclusion, the use of organotypic cultures permits predictions of neuroactive properties of newly designed drugs on an intermediate level, and should therefore open up avenues for a more creative and economic drug development process.

Keywords: Organotypic cultures, anaesthetics, cortex, spinal cord, GABA(A) receptor, benzodiazepines, immobilizing effects, Metabotropic GABAB receptors, ionotropic GABAA receptors, amnestic, allosteric binding site, amygdala, hippocampus, GABAergic interneurons, dopaminergic cells, brain stem, cerebellum, neocortex, Pyramidal neurons, thalamic axon, cholinergic neurons, human embryo kidney (HEK), antiepileptic, propofol, local field potential (LFP), electroencenphalogram (EEG), synchronizing, rocuronium, mivacurium, bicuculline, potassium channels, inhibitory postsynaptic currents, extrasynaptic sites, phosphorylation, neurotransmitters