Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
In situ calibration and [H+] sensitivity of the fluorescent Na+ indicator SBFI
Autore:
Diarra, A; Sheldon, C; Church, J;
Indirizzi:
Univ British Columbia, Dept Anat, Vancouver, BC V6T 1Z3, Canada Univ British Columbia Vancouver BC Canada V6T 1Z3 ver, BC V6T 1Z3, Canada
Titolo Testata:
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY
fascicolo: 6, volume: 280, anno: 2001,
pagine: C1623 - C1633
SICI:
0363-6143(200106)280:6<C1623:ISCA[S>2.0.ZU;2-E
Fonte:
ISI
Lingua:
ENG
Soggetto:
RAT HIPPOCAMPAL-NEURONS; BINDING BENZOFURAN ISOPHTHALATE; CEREBELLAR GRANULE CELLS; INTRACELLULAR SODIUM; CYTOSOLIC SODIUM; VENTRICULAR MYOCYTES; GLUCOSE DEPRIVATION; ACID EXTRUSION; GLIAL-CELL; PH;
Keywords:
sodium-binding benzofuran isophthalate; hippocampal neuron; intracellular sodium; intracellular pH;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
40
Recensione:
Indirizzi per estratti:
Indirizzo: Church, J Univ British Columbia, Dept Anat, 2177 Wesbrook Mall, Vancouver,BC V6T 1Z3, Canada Univ British Columbia 2177 Wesbrook Mall Vancouver BC Canada V6T 1Z3
Citazione:
A. Diarra et al., "In situ calibration and [H+] sensitivity of the fluorescent Na+ indicator SBFI", AM J P-CELL, 280(6), 2001, pp. C1623-C1633

Abstract

Despite the popularity of Na+-binding benzofuran isophthalate (SBFI) to measure intracellular free Na+ concentrations ([Na+](i)), the in situ calibration techniques described to date do not favor the straightforward determination of all of the constants required by the standard equation (Grynkiewicz G, Poenie M, and Tsien RY. J Biol Chem 260: 3440-3450, 1985) to convert the ratiometric signal into [Na+]. We describe a simple method in which SBFIratio values obtained during a "full" in situ calibration are fit by a three-parameter hyperbolic equation; the apparent dissociation constant (K-d) of SBFI for Na+ can then be resolved by means of a three-parameter hyperbolic decay equation. We also developed and tested a "one-point" technique forcalibrating SBFI ratios in which the ratio value obtained in a neuron at the end of an experiment during exposure to gramicidin D and 10 mM Na+ is used as a normalization factor for ratios obtained during the experiment; each normalized ratio is converted to [Na+](i) using a modification of the standard equation and parameters obtained from a full calibration. Finally, weextended the characterization of the pH dependence of SBFI in situ. Although the K-d of SBFI for Na+ was relatively insensitive to changes in pH in the range 6.8-7.8, acidification resulted in an apparent decrease, and alkalinization in an apparent increase, in [Na+](i) values. The magnitudes of the apparent changes in [Na+](i) varied with absolute [Na+](i), and a method was developed for correcting [Na+](i) values measured with SBFI for changesin intracellular pH.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 02/04/20 alle ore 12:43:21