title: High-resolution spatio-temporal bioactivity of a novel peptide revealed by optical imaging in rat orbitofrontal cortex in vitro: possible implications for neurodegenerative diseases.
abstract: Acetylcholinesterase (AChE) is now well known to have a secondary, non-enzymatic function independent of cholinergic transmission. In the last decade, the part of the molecule responsible for this action has been identified, i.e. a 14 amino acid peptide fragment ('T14'), deriving from the C-terminus of AChE: this peptide has been shown to be bioactive in a range of preparations and to act at an allosteric site on α₇ nicotinic acetylcholine receptors (α₇-nAChRs). Of particular significance is the finding that AChE-related peptides trigger calcium-induced neurotoxicity that may be pivotal in the process of neurodegenerative diseases, such as Alzheimer's. However to date all studies have been performed on isolated cell preparations. The aim of this study was therefore to characterise the bioactivity of T14 on meso-scale in vitro cortical networks ('neuronal assemblies') from rat brain slices containing orbitofrontal cortex. Local field potential (LFP) recordings showed that the T14 peptide has a selective, holistic action on cortical networks in a modulatory biphasic manner i.e. predisposing excitation at concentrations of up to 1 μM, after which the trend is reversed in favour of inhibition at higher doses (>1 μM). By contrast, a scrambled variant of the T14 peptide sequence (S14), showed no significant changes in neuronal activation. Optical imaging using voltage-sensitive dyes (VSDI) corroborated the electrophysiological findings and also provided further insight into the spatial dynamics of the effects of the peptide: T14 application had a facilitatory effect i.e. increased the time-course of activation at sub-micromolar concentrations only (700 nM) without significantly affecting the spread of evoked assemblies. Moreover: co-applying T14 with the α₇-nAChR competitive antagonist methyllycaconitine (MLA) produced inhibition in activation synchrony not seen with either agent on their own, suggesting an additive inhibitory effect. In conclusion, the T14 peptide derived from AChE produced a dose-dependent biphasic modulation of cortical networks activity dependent on the α₇-nAChR: these findings should thus provide a more comprehensive insight into the immediate actions of a novel bioactive agent of high potential relevance to neurodegenerative disorders such as Alzheimer's disease.
authors.affiliation: Department of Pharmacology, University of Oxford, Mansfield Road, Oxford OX1 3QT, United Kingdom. scott.badin@seh.ox.ac.uk
authors.forename: Antoine-Scott
authors.initials: AS
authors.lastname: Badin
authors.forename: John
authors.initials: J
authors.lastname: Eraifej
authors.forename: Susan
authors.initials: S
authors.lastname: Greenfield
mesh_terms: D000110:Acetylcholinesterase / Q000737:chemistry*; D000157:Aconitine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D008297:Male; D019636:Neurodegenerative Diseases / Q000209:etiology / Q000378:metabolism; D061848:Optical Imaging; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology*; D017397:Prefrontal Cortex / Q000187:drug effects* / Q000378:metabolism* / Q000502:physiology; D051381:Rats; D064569:alpha7 Nicotinic Acetylcholine Receptor / Q000819:agonists* / Q000037:antagonists & inhibitors
chemical_list: D010446:Peptide Fragments; D064569:alpha7 Nicotinic Acetylcholine Receptor; C054634:methyllycaconitine; D000110:Acetylcholinesterase; D000157:Aconitine
keywords: Acetylcholinesterase (AChE); Alzheimer's disease; C-terminal peptide; Field potentials; Methyllycaconitine (MLA); Optical imaging; Orbitofrontal cortex; Rat brain slices; α(7)-Nicotinic receptor