Dataset Information

Direct whole-cell patch-clamp recordings from small boutons in rodent primary neocortical neuron cultures.

ABSTRACT: Direct electrical recordings from conventional boutons in the mammalian central nervous system have proven challenging due to their small size. Here, we provide a protocol for direct whole-cell patch-clamp recordings from small presynaptic boutons of primary dissociated cultured neurons of the rodent neocortex. We describe steps to prepare primary neocortical cultures and recording pipettes, followed by identifying boutons and establishing a whole-cell bouton recording. We then provide details on precise pipette capacitance compensation required for high-resolution current-clamp recordings from boutons. For further details on the use and execution of this protocol, please refer to Ritzau-Jost et al.¹.

SUBMITTER: Ritzau-Jost A

PROVIDER: S-EPMC10026040 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature

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Direct whole-cell patch-clamp recordings from small boutons in rodent primary neocortical neuron cultures.

Ritzau-Jost Andreas A Nerlich Jana J Kaas Thomas T Krueger Martin M Tsintsadze Timur T Eilers Jens J Barbour Boris B Smith Stephen M SM Hallermann Stefan S

STAR protocols 20230314 2

Direct electrical recordings from conventional boutons in the mammalian central nervous system have proven challenging due to their small size. Here, we provide a protocol for direct whole-cell patch-clamp recordings from small presynaptic boutons of primary dissociated cultured neurons of the rodent neocortex. We describe steps to prepare primary neocortical cultures and recording pipettes, followed by identifying boutons and establishing a whole-cell bouton recording. We then provide details o ...[more]

PMID: 36920913

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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

Direct whole-cell patch-clamp recordings from small boutons in rodent primary neocortical neuron cultures.

Publications

Direct whole-cell patch-clamp recordings from small boutons in rodent primary neocortical neuron cultures.

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