Dataset Information

Comparison of biophysical properties of ?1?2 and ?3?2 GABAA receptors in whole-cell patch-clamp electrophysiological recordings.

ABSTRACT: In the present study we have characterized the biophysical properties of wild-type (WT) ?1?2 and ?3?2 GABAA receptors and probed the molecular basis for the observed differences. The activation and desensitization behavior and the residual currents of the receptors expressed in HEK293 cells were determined in whole-cell patch clamp recordings. Kinetic parameters of ?1?2 and ?3?2 activation differed significantly, with ?1?2 and ?3?2 exhibiting rise times (10-90%) of 24 ± 2 ms and 51 ± 7 ms, respectively. In contrast, the two receptors exhibited largely comparable desensitization behavior with decay currents that could be fitted to exponential functions with two or three components. Most notably, the two receptor compositions displayed different degrees of desentization, with the residual currents of ?1?2 and ?3?2 constituting 34 ± 2% and 21 ± 2% of the peak current, respectively. The respective contributions of the extracellular domains and the transmembrane/intracellular domains of the ?-subunit to these physiological profiles were next assessed in recordings from cells expressing ??2 receptors comprising chimeric ?-subunits. The rise times displayed by ?1ECD/?3TMD?2 and ?3ECD/?1TMD?2 receptors were intermediate to those of WT ?1?2 and WT ?3?2, and the distribution of the different components of the current decays exhibited by the two chimeric receptors followed the same pattern as the two WT receptors. The residual current exhibited by ?1ECD/?3TMD?2 (23 ± 3%) was similar to that of ?3?2 but significantly different from that of ?1?2, whereas the residual current displayed by ?3ECD/?1TMD?2 (27 ± 2%) was intermediate to and did not differ significantly from either of the WT receptors. This points to molecular differences in the transmembrane/intracellular domains of the ?-subunit as the main determinants of the observed differences in receptor physiology between ?1?2 and ?3?2 receptors.

SUBMITTER: Olander ER

PROVIDER: S-EPMC7263626 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Comparison of biophysical properties of α1β2 and α3β2 GABAA receptors in whole-cell patch-clamp electrophysiological recordings.

Olander Emma Rie ER Janzen Dieter D Villmann Carmen C Jensen Anders A AA

PloS one 20200601 6

In the present study we have characterized the biophysical properties of wild-type (WT) α1β2 and α3β2 GABAA receptors and probed the molecular basis for the observed differences. The activation and desensitization behavior and the residual currents of the receptors expressed in HEK293 cells were determined in whole-cell patch clamp recordings. Kinetic parameters of α1β2 and α3β2 activation differed significantly, with α1β2 and α3β2 exhibiting rise times (10-90%) of 24 ± 2 ms and 51 ± 7 ms, respe ...[more]

PMID: 32479525

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Project description:A lot of efforts have been made to understand the structure and function of neocortical circuits. In fact, a promising way to understand the functions of cortical circuits is the classification of the neural types, based on their different properties. Recent studies focused on applying modern computational methods to classify neurons based on molecular, morphological, physiological, or mixed of these criteria. Although there are studies in the literature on in vitro/vivo extracellular or in vitro intracellular recordings, a study on the classification of neuronal types using in vivo whole-cell patch-clamp recordings is still lacking. We thus proposed a novel semi-supervised classification method based on waveform shape of neurons' spikes using in vivo whole-cell patch-clamp recordings. We, first, detected spike candidates. Then discriminative features were extracted from the time samples of the spikes using discrete cosine transform. We then extracted the center of clusters using fuzzy c-mean clustering and finally, the neurons were classified using the minimum distance classifier. We distinguished three types of neurons: excitatory pyramidal cells (Pyr) and two types of inhibitory neurons: GABAergic- parvalbumin positive (PV), and somatostatin positive (SST) non-pyramidal cells in layer II/III of the mice primary visual cortex. We used 10-fold cross validation in our study. The classification accuracy for PV, Pyr, and SST was 91.59 ± 1.69, 97.47 ± 0.67, and 89.06 ± 1.99, respectively. Overall, the algorithm correctly classified 92.67 ± 0.54% of the cells, confirming the relative robustness of the discriminant functions. The performance of the method was further assessed on in vitro recordings by using a pool of 50 neurons from Allen institute Cell Types Database (5 major subtypes of neurons: Pyr, PV, SST, 5HT3a, and vasoactive intestinal peptide (VIP) cells). Its overall accuracy was 84.13 ± 0.81% on this data set using cross validation framework. The proposed algorithm is thus a promising new tool in recognizing cell's type with high accuracy in laboratories using in vivo/vitro whole-cell patch-clamp recording technique. The developed programs and the entire dataset are available online to interested readers.

Dataset Information

Comparison of biophysical properties of ?1?2 and ?3?2 GABAA receptors in whole-cell patch-clamp electrophysiological recordings.

Publications

Comparison of biophysical properties of α1β2 and α3β2 GABAA receptors in whole-cell patch-clamp electrophysiological recordings.

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