PMID- 7810712
OWN - NLM
STAT- MEDLINE
DCOM- 19950127
LR  - 20171213
IS  - 0002-9513 (Print)
IS  - 0002-9513 (Linking)
VI  - 267
IP  - 6 Pt 2
DP  - 1994 Dec
TI  - Use of MQAE for measurement of intracellular [Cl-] in cultured aortic smooth 
      muscle cells.
PG  - H2114-23
AB  - A novel fluorescent indicator, N-[ethoxycarbonylmethyl]-6-methoxy-quinolinium 
      bromide (MQAE), was used to measure intracellular chloride concentration ([Cl-]i) 
      in primary cultures of rat aortic smooth muscle cells (VSMC). The hydrolytic and 
      fluorescent properties of the dye were characterized. The intracellular 
      Stern-Volmer constant was calculated to be 25 M-1. Cl- efflux curves were 
      characteristic of saturation-type kinetics, with an apparent Michaelis-Menten 
      constant value of 11 +/- 4.8 (SD) mM, a maximum velocity of 0.038 +/- 0.021 mM/s, 
      and a half time (t1/2) of 9.0 +/- 3.7 min. The average efflux rate in the first 
      10 min (0.023 +/- 0.004 mM/s) was reduced in the presence of either 130 microM 
      4,4'-diisothiocyanato-dihydrostilbene-2,2'-disulfonic acid (H2DIDS) (0.014 +/- 
      0.006, P = 0.02) or 40 microM furosemide (0.017 +/- 0.004, P = 0.04). Restoration 
      of physiological extracellular chloride concentration ([Cl-]o) after zero Cl- 
      resulted in net Cl- influx with a t1/2 of 3.6 +/- 1.0 min. The initial Cl- influx 
      rate was reduced after exposure to furosemide, from 0.069 +/- 0.006 to 0.046 +/- 
      0.008 mM/s, P < 0.002, and was reduced after exposure to H2DIDS from 0.102 +/- 
      0.013 to 0.033 +/- 0.003 mM/s, P < 0.001. Furosemide reduced the steady-state 
      [Cl-]i from 31.6 +/- 3.2 to 26.1 +/- 2.4 mM, P < 0.01, whereas H2DIDS had little 
      effect on [Cl-]i. Our results demonstrate that MQAE can be used to measure [Cl-]i 
      in primary cultures of VSMC.
FAU - Koncz, C
AU  - Koncz C
AD  - Department of Medicine, University of Illinois College of Medicine at Chicago.
FAU - Daugirdas, J T
AU  - Daugirdas JT
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Am J Physiol
JT  - The American journal of physiology
JID - 0370511
RN  - 0 (Chlorides)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Quinolinium Compounds)
RN  - 18010-05-4 (N-(ethoxycarbonylmethyl)-6-methoxyquinolinium)
RN  - 7LXU5N7ZO5 (Furosemide)
RN  - Q1O6DSW23R (4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid)
SB  - IM
MH  - 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid/pharmacology
MH  - Animals
MH  - Aorta, Thoracic
MH  - Cells, Cultured
MH  - Chlorides/*analysis/chemistry/metabolism
MH  - *Fluorescent Dyes
MH  - Furosemide/pharmacology
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - Male
MH  - Muscle, Smooth, Vascular/*chemistry
MH  - Osmolar Concentration
MH  - Quinolinium Compounds/chemistry/metabolism
MH  - Rats
MH  - Spectrometry, Fluorescence
MH  - Temperature
MH  - Viscosity
EDAT- 1994/12/01 00:00
MHDA- 1994/12/01 00:01
CRDT- 1994/12/01 00:00
PHST- 1994/12/01 00:00 [pubmed]
PHST- 1994/12/01 00:01 [medline]
PHST- 1994/12/01 00:00 [entrez]
AID - 10.1152/ajpheart.1994.267.6.H2114 [doi]
PST - ppublish
SO  - Am J Physiol. 1994 Dec;267(6 Pt 2):H2114-23. doi: 
      10.1152/ajpheart.1994.267.6.H2114.