Fluorescent detection of Ca2+-permeable AMPA/kainate receptor activation in murine neocortical neurons

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Agonist stimulated Co2+ uptake is used to identify neurons expressing Ca2+-permeable AMPA/kaniate receptors (Ca-A/K receptors). Based on the selective permeability of Co2+ through these receptors we have developed a fluorometric method that utilizes the fluorescence laser imaging plate reader (FLIPR). We used the dye calcein whose fluorescence is stoichiometrically quenched by Co2+, while being only minimally affected by variations in intracellular Ca2+. Application of AMPA in the presence of cyclothiazide led to a concentration-dependent increase in Co2+ uptake in the neocortical neurons. Similar concentration- dependent increments in Co2+ uptake were observed with kainate treatment. 2,3-Dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7- sulfonamide (NBQX), an AMPA/kainate receptor antagonist, blocked the AMPA-induced Co2+ influx. The fluorometric method described affords a rapid, high throughput and quantitative procedure for investigation of Ca-A/K receptors in intact neurons.

Original languageEnglish
Pages (from-to)145-148
Number of pages4
JournalNeuroscience Letters
Volume351
Issue number3
DOIs
StatePublished - Nov 20 2003
Externally publishedYes

Fingerprint

Kainic Acid Receptors
AMPA Receptors
alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid
Neurons
Quinoxalines
Kainic Acid
Optical Imaging
Sulfonamides
Permeability
Lasers
Coloring Agents
Fluorescence

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

Cite this

@article{d40f19da7567470cbef22564a66795b2,
title = "Fluorescent detection of Ca2+-permeable AMPA/kainate receptor activation in murine neocortical neurons",
abstract = "Agonist stimulated Co2+ uptake is used to identify neurons expressing Ca2+-permeable AMPA/kaniate receptors (Ca-A/K receptors). Based on the selective permeability of Co2+ through these receptors we have developed a fluorometric method that utilizes the fluorescence laser imaging plate reader (FLIPR). We used the dye calcein whose fluorescence is stoichiometrically quenched by Co2+, while being only minimally affected by variations in intracellular Ca2+. Application of AMPA in the presence of cyclothiazide led to a concentration-dependent increase in Co2+ uptake in the neocortical neurons. Similar concentration- dependent increments in Co2+ uptake were observed with kainate treatment. 2,3-Dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7- sulfonamide (NBQX), an AMPA/kainate receptor antagonist, blocked the AMPA-induced Co2+ influx. The fluorometric method described affords a rapid, high throughput and quantitative procedure for investigation of Ca-A/K receptors in intact neurons.",
author = "Dravid, {Shashank M.} and Murray, {Thomas F.}",
year = "2003",
month = "11",
day = "20",
doi = "10.1016/j.neulet.2003.07.017",
language = "English",
volume = "351",
pages = "145--148",
journal = "Neuroscience Letters",
issn = "0304-3940",
publisher = "Elsevier Ireland Ltd",
number = "3",

}

TY - JOUR

T1 - Fluorescent detection of Ca2+-permeable AMPA/kainate receptor activation in murine neocortical neurons

AU - Dravid, Shashank M.

AU - Murray, Thomas F.

PY - 2003/11/20

Y1 - 2003/11/20

N2 - Agonist stimulated Co2+ uptake is used to identify neurons expressing Ca2+-permeable AMPA/kaniate receptors (Ca-A/K receptors). Based on the selective permeability of Co2+ through these receptors we have developed a fluorometric method that utilizes the fluorescence laser imaging plate reader (FLIPR). We used the dye calcein whose fluorescence is stoichiometrically quenched by Co2+, while being only minimally affected by variations in intracellular Ca2+. Application of AMPA in the presence of cyclothiazide led to a concentration-dependent increase in Co2+ uptake in the neocortical neurons. Similar concentration- dependent increments in Co2+ uptake were observed with kainate treatment. 2,3-Dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7- sulfonamide (NBQX), an AMPA/kainate receptor antagonist, blocked the AMPA-induced Co2+ influx. The fluorometric method described affords a rapid, high throughput and quantitative procedure for investigation of Ca-A/K receptors in intact neurons.

AB - Agonist stimulated Co2+ uptake is used to identify neurons expressing Ca2+-permeable AMPA/kaniate receptors (Ca-A/K receptors). Based on the selective permeability of Co2+ through these receptors we have developed a fluorometric method that utilizes the fluorescence laser imaging plate reader (FLIPR). We used the dye calcein whose fluorescence is stoichiometrically quenched by Co2+, while being only minimally affected by variations in intracellular Ca2+. Application of AMPA in the presence of cyclothiazide led to a concentration-dependent increase in Co2+ uptake in the neocortical neurons. Similar concentration- dependent increments in Co2+ uptake were observed with kainate treatment. 2,3-Dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7- sulfonamide (NBQX), an AMPA/kainate receptor antagonist, blocked the AMPA-induced Co2+ influx. The fluorometric method described affords a rapid, high throughput and quantitative procedure for investigation of Ca-A/K receptors in intact neurons.

UR - http://www.scopus.com/inward/record.url?scp=0242499507&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0242499507&partnerID=8YFLogxK

U2 - 10.1016/j.neulet.2003.07.017

DO - 10.1016/j.neulet.2003.07.017

M3 - Article

C2 - 14623127

AN - SCOPUS:0242499507

VL - 351

SP - 145

EP - 148

JO - Neuroscience Letters

JF - Neuroscience Letters

SN - 0304-3940

IS - 3

ER -