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NMDA receptor activity in circulating red blood cells: methods of detection


Makhro, Asya; Kaestner, Lars; Bogdanova, Anna (2017). NMDA receptor activity in circulating red blood cells: methods of detection. In: Burnashev, Nail; Szepetowski, Pierre. NMDA Receptors. Hatfield, Hertfordshire, UK: Springer, 265-282.

Abstract

Abundance and activity of N-methyl-D-aspartate (NMDA) in circulating red blood cells contributes to the maintenance of intracellular Ca(2+) in these cells and, by doing that, controls red cell volume, membrane stability, and O2 carrying capacity. Detection of the NMDA receptor activity in red blood cells is challenging as the number of its copies is low and shows substantial cell-to-cell heterogeneity. Receptor abundance is reliably assessed using the radiolabeled antagonist ([(3)H]MK-801) binding technique. Uptake of Ca(2+) following the NMDA receptor activation is detected in cells loaded with Ca(2+)-sensitive fluorescent dye Fluo-4 AM. Both microfluorescence live-cell imaging and flow cytometry may be used for fluorescence intensity detection. Automated patch clamp is currently used for recording of electric currents triggered by the stimulation of the NMDA receptor. These currents are mediated by the Ca(2+)-sensitive K(+) (Gardos) channels that open upon Ca(2+) uptake via the active NMDA receptor. Furthermore, K(+) flux through the Gardos channels induced by the NMDA receptor stimulation in red blood cells may be detected using unidirectional K(+)((86)Rb(+)) influx.

Abstract

Abundance and activity of N-methyl-D-aspartate (NMDA) in circulating red blood cells contributes to the maintenance of intracellular Ca(2+) in these cells and, by doing that, controls red cell volume, membrane stability, and O2 carrying capacity. Detection of the NMDA receptor activity in red blood cells is challenging as the number of its copies is low and shows substantial cell-to-cell heterogeneity. Receptor abundance is reliably assessed using the radiolabeled antagonist ([(3)H]MK-801) binding technique. Uptake of Ca(2+) following the NMDA receptor activation is detected in cells loaded with Ca(2+)-sensitive fluorescent dye Fluo-4 AM. Both microfluorescence live-cell imaging and flow cytometry may be used for fluorescence intensity detection. Automated patch clamp is currently used for recording of electric currents triggered by the stimulation of the NMDA receptor. These currents are mediated by the Ca(2+)-sensitive K(+) (Gardos) channels that open upon Ca(2+) uptake via the active NMDA receptor. Furthermore, K(+) flux through the Gardos channels induced by the NMDA receptor stimulation in red blood cells may be detected using unidirectional K(+)((86)Rb(+)) influx.

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Additional indexing

Item Type:Book Section, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Institute of Veterinary Physiology
Dewey Decimal Classification:570 Life sciences; biology
Date:12 September 2017
Deposited On:17 Oct 2017 15:55
Last Modified:30 Mar 2018 04:02
Publisher:Springer
ISSN:1064-3745
ISBN:978-1-4939-7320-0
OA Status:Closed
Publisher DOI:https://doi.org/10.1007/978-1-4939-7321-7_15
Related URLs:http://ww.springer.com/de/book/9781493973200 (Publisher)
PubMed ID:28986879
Project Information:
  • : FunderFP7
  • : Grant ID602121
  • : Project TitleCOMMITMENT - Combined Molecular Microscopy for Therapy and Personalized Medication in Rare Anaemias Treatments
  • : FunderH2020
  • : Grant ID675115
  • : Project TitleRELEVANCE - Regulation of red cell life-span, erythropoiesis, survival, senescence and clearance

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