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An automated planar patch-clamp approach to measure the membrane potential and resting membrane currents in a human cerebrovascular endothelial cell line.
Soda, Teresa; Negri, Sharon; Scarpellino, Giorgia; Berra-Romani, Roberto; De Sarro, Giovambattista; Moccia, Francesco; Brunetti, Valentina.
Afiliación
  • Soda T; Department of Health Science, University Magna Graecia of Catanzaro, Catanzaro 88100, Italy.
  • Negri S; Laboratory of General Physiology, Department of Biology and Biotechnology "L. Spallanzani", University of Pavia, Pavia 27100, Italy.
  • Scarpellino G; Laboratory of General Physiology, Department of Biology and Biotechnology "L. Spallanzani", University of Pavia, Pavia 27100, Italy.
  • Berra-Romani R; Department of Biomedicine, School of Medicine, Benemérita Universidad Autónoma de Puebla, Puebla 72410, Mexico.
  • De Sarro G; Department of Health Science, University Magna Graecia of Catanzaro, Catanzaro 88100, Italy.
  • Moccia F; Laboratory of General Physiology, Department of Biology and Biotechnology "L. Spallanzani", University of Pavia, Pavia 27100, Italy. Electronic address: francesco.moccia@unimol.it.
  • Brunetti V; Laboratory of General Physiology, Department of Biology and Biotechnology "L. Spallanzani", University of Pavia, Pavia 27100, Italy.
J Neurosci Methods ; 410: 110248, 2024 Oct.
Article en En | MEDLINE | ID: mdl-39117152
ABSTRACT

BACKGROUND:

The conventional "whole-cell patch-clamp" recording technique is widely used to measure the resting membrane potential (VM) and to dissect the underlying membrane ionic conductances in isolated vascular endothelial cells. NEW

METHOD:

Herein, we assessed whether the automated patch-clamp (APC) technology, which replaces the traditional patch-pipette with a planar substrate to permit researchers lacking formal training in electrophysiology to generate large amounts of data in a relatively short time, can be used to characterize the bioelectrical activity of vascular endothelial cells. We assessed whether the Port-a-Patch planar patch-clamp system, which is regarded as the smallest electrophysiological rig available on the market, can be used to measure the VM and resting membrane currents in the human cerebrovascular endothelial cell line, hCMEC/D3. COMPARISON WITH EXISTING

METHODS:

We demonstrated that the Port-a-Patch planar patch-clamp system provides the same values of the resting VM as those provided by the conventional patch-clamp technique. Furthermore, the APC technology provides preliminary data demonstrating that the resting VM of hCMEC/D3 cells is primarily contributed by Cl- and Na+, as demonstrated with the patch-clamp technique for many other endothelial cell types.

CONCLUSIONS:

The Port-a-Patch planar patch-clamp system can be successfully used to measure the resting VM and the underlying membrane ionic conductances in hCMEC/D3 cells. We envisage that this easy-to-use APC system could also be extremely useful for the investigation of the membrane currents that can be activated by chemical, thermal, optical, and mechanical stimuli in this cell line as well as in other types of isolated vascular endothelial cells.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas de Placa-Clamp / Células Endoteliales / Potenciales de la Membrana Límite: Humans Idioma: En Revista: J Neurosci Methods Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas de Placa-Clamp / Células Endoteliales / Potenciales de la Membrana Límite: Humans Idioma: En Revista: J Neurosci Methods Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos