Dataset Information

Pharmacological characterization of a non-inactivating outward current observed in mouse cerebellar Purkinje neurones.

ABSTRACT: Whole-cell patch clamp recordings were used to investigate the properties of a non-inactivating outward current observed in mouse cerebellar Purkinje neurones at a holding potential of -20 mV. Increasing the external potassium (K(+)) concentration from 3 mM to 20 mM produced a rightward shift in the observed reversal potential of approximately 30 mV or approximately 40 mV for a K(+)-or a caesium (Cs(+))-based intracellular solution respectively, indicating the outward current was a K(+) current. The outward current was partially inhibited by the K(+) channel blocker, tetraethylammonium (TEA; IC(50)=0.15 mM). Subsequently, the background or TEA-insensitive current was measured in the presence of 1 mM TEA. The background current was reversibly inhibited by barium (Ba(2+); 300 microM, 50%) and potentiated by the application of arachidonic acid (AA; 1 mM, 62%). The volatile anaesthetic, halothane (1 mM), and the neuroprotectant, riluzole (500 microM), both reversibly inhibited the background current by 54% and 36% respectively. The background current was insensitive to changes in both intracellular and extracellular acidification. The GABA(B) and mu-opioid receptor agonists, baclofen and [D-Ala(2), N-MePhe(4)-Gly-ol(5)] enkephalin (DAMGO) both reversibly potentiated the outward current by 42% and 26% respectively. In contrast, the metabotropic glutamate receptor and acetylcholine receptor agonists, (S)-3,5-dihydroxyphenylglycine (DHPG) and muscarine both reversibly inhibited the outward current by 48% and 42% respectively. These data suggest that cerebellar Purkinje neurones possess a background current which shares several properties with recently cloned two-pore K(+) channels, particularly THIK-1.

SUBMITTER: Bushell T

PROVIDER: S-EPMC1573182 | biostudies-literature | 2002 Feb

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Pharmacological characterization of a non-inactivating outward current observed in mouse cerebellar Purkinje neurones.

Bushell Trevor T Clarke Catherine C Mathie Alistair A Robertson Brian B

British journal of pharmacology 20020201 3

Whole-cell patch clamp recordings were used to investigate the properties of a non-inactivating outward current observed in mouse cerebellar Purkinje neurones at a holding potential of -20 mV. Increasing the external potassium (K(+)) concentration from 3 mM to 20 mM produced a rightward shift in the observed reversal potential of approximately 30 mV or approximately 40 mV for a K(+)-or a caesium (Cs(+))-based intracellular solution respectively, indicating the outward current was a K(+) current. ...[more]

PMID: 11834618

Similar Datasets

Project description:Despite our fine-grain anatomical knowledge of the cerebellar cortex, electrophysiological studies of circuit information processing over the last fifty years have been hampered by the difficulty of reliably assigning signals to identified cell types. We approached this problem by assessing the spontaneous activity signatures of identified cerebellar cortical neurones. A range of statistics describing firing frequency and irregularity were then used, individually and in combination, to build Gaussian Process Classifiers (GPC) leading to a probabilistic classification of each neurone type and the computation of equi-probable decision boundaries between cell classes. Firing frequency statistics were useful for separating Purkinje cells from granular layer units, whilst firing irregularity measures proved most useful for distinguishing cells within granular layer cell classes. Considered as single statistics, we achieved classification accuracies of 72.5% and 92.7% for granular layer and molecular layer units respectively. Combining statistics to form twin-variate GPC models substantially improved classification accuracies with the combination of mean spike frequency and log-interval entropy offering classification accuracies of 92.7% and 99.2% for our molecular and granular layer models, respectively. A cross-species comparison was performed, using data drawn from anaesthetised mice and decerebrate cats, where our models offered 80% and 100% classification accuracy. We then used our models to assess non-identified data from awake monkeys and rabbits in order to highlight subsets of neurones with the greatest degree of similarity to identified cell classes. In this way, our GPC-based approach for tentatively identifying neurones from their spontaneous activity signatures, in the absence of an established ground-truth, nonetheless affords the experimenter a statistically robust means of grouping cells with properties matching known cell classes. Our approach therefore may have broad application to a variety of future cerebellar cortical investigations, particularly in awake animals where opportunities for definitive cell identification are limited.

Project description:RationaleA chromosomal haplotype producing cardiac overexpression of dipeptidyl peptidase-like protein-6 (DPP6) causes familial idiopathic ventricular fibrillation. The molecular basis of transient outward current (I(to)) in Purkinje fibers (PFs) is poorly understood. We hypothesized that DPP6 contributes to PF I(to) and that its overexpression might specifically alter PF I(to) properties and repolarization.ObjectiveTo assess the potential role of DPP6 in PF I(to).Methods and resultsClinical data in 5 idiopathic ventricular fibrillation patients suggested arrhythmia origin in the PF-conducting system. PF and ventricular muscle I(to) had similar density, but PF I(to) differed from ventricular muscle in having tetraethylammonium sensitivity and slower recovery. DPP6 overexpression significantly increased, whereas DPP6 knockdown reduced, I(to) density and tetraethylammonium sensitivity in canine PF but not in ventricular muscle cells. The K(+)-channel interacting β-subunit K(+)-channel interacting protein type-2, essential for normal expression of I(to) in ventricular muscle, was weakly expressed in human PFs, whereas DPP6 and frequenin (neuronal calcium sensor-1) were enriched. Heterologous expression of Kv4.3 in Chinese hamster ovary cells produced small I(to); I(to) amplitude was greatly enhanced by coexpression with K(+)-channel interacting protein type-2 or DPP6. Coexpression of DPP6 with Kv4.3 and K(+)-channel interacting protein type-2 failed to alter I(to) compared with Kv4.3/K(+)-channel interacting protein type-2 alone, but DPP6 expression with Kv4.3 and neuronal calcium sensor-1 (to mimic PF I(to) composition) greatly enhanced I(to) compared with Kv4.3/neuronal calcium sensor-1 and recapitulated characteristic PF kinetic/pharmacological properties. A mathematical model of cardiac PF action potentials showed that I(to) enhancement can greatly accelerate PF repolarization.ConclusionsThese results point to a previously unknown central role of DPP6 in PF I(to), with DPP6 gain of function selectively enhancing PF current, and suggest that a DPP6-mediated PF early-repolarization syndrome might be a novel molecular paradigm for some forms of idiopathic ventricular fibrillation.

Dataset Information

Pharmacological characterization of a non-inactivating outward current observed in mouse cerebellar Purkinje neurones.

Publications

Pharmacological characterization of a non-inactivating outward current observed in mouse cerebellar Purkinje neurones.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets