PMID- 16630555
OWN - NLM
STAT- MEDLINE
DCOM- 20060622
LR  - 20131121
IS  - 0006-291X (Print)
IS  - 0006-291X (Linking)
VI  - 344
IP  - 2
DP  - 2006 Jun 2
TI  - Simultaneous measurement of superoxide generation and intracellular Ca2+ 
      concentration reveals the effect of extracellular Ca2+ on rapid and transient 
      contents of superoxide generation in differentiated THP-1 cells.
PG  - 571-80
AB  - We invented a simultaneous measuring instrument of fluorescence and 
      chemiluminescence, realizing the analysis of chronological correlation between 
      change in intracellular Ca2+ concentration ([Ca2+]i) and superoxide generation. A 
      human monocytic cell line, THP-1, differentiated to be neutrophil-like cells 
      generated superoxide with increase in intracellular Ca2+ concentration when 
      stimulated with formyl-methionyl-leucyl-phenylalanine (fMLP) whereas PMA, phorbol 
      ester-stimulated superoxide response occurred without change in [Ca2+]i. The 
      cells treated with TMB-8, an intracellular Ca2+ antagonist, generated superoxide 
      rapidly as well as transiently with transient [Ca2+]i elevation after stimulation 
      with fMLP, whereas EGTA-treated cells generated superoxide slowly as well as 
      persistently with transient [Ca2+]i elevation after the stimulation. These 
      results suggest that the rapid and transient contents of superoxide generation 
      are specific for Ca2+ influx from the extracellular domain. Verapamil, 
      voltage-dependent Ca2+ channel blocker, dose-dependently inhibited 
      fMLP-stimulated extracellular Ca2+ influx and superoxide generation without 
      affecting PMA-stimulated superoxide generation. Other channel blockers tested, 
      nifedipine and diltiazem, similarly inhibited these fMLP-stimulated responses. 
      Numerical analysis of the values of the response curves elucidated that TMB-8 or 
      the channel blocker reveals or eliminates the same contents of superoxide 
      generation by the antagonism of intracellular Ca2+ release or extracellular Ca2+ 
      influx, respectively. Taking these results together, the characteristic 
      extracellular Ca2+ influx essential for superoxide generation was first revealed 
      by the simultaneous measurement of superoxide generation and change in [Ca2+]i.
FAU - Ishibashi, Kaname
AU  - Ishibashi K
AD  - Laboratory of Molecular Biophotonics, 5000 Hirakuchi, Hamamatsu 434-8555, Japan. 
      kaname.ishibashi@ims.jti.co.jp
FAU - Okazaki, Shigetoshi
AU  - Okazaki S
FAU - Hiramatsu, Mitsuo
AU  - Hiramatsu M
LA  - eng
PT  - Journal Article
DEP - 20060310
PL  - United States
TA  - Biochem Biophys Res Commun
JT  - Biochemical and biophysical research communications
JID - 0372516
RN  - 11062-77-4 (Superoxides)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Calcium/*metabolism
MH  - Calcium Signaling/*physiology
MH  - Cell Culture Techniques/methods
MH  - Cell Differentiation
MH  - Cell Line
MH  - Extracellular Fluid
MH  - Humans
MH  - Intracellular Fluid/*metabolism
MH  - Kinetics
MH  - Metabolic Clearance Rate
MH  - Monocytes/drug effects/*metabolism
MH  - Superoxides/*metabolism
EDAT- 2006/04/25 09:00
MHDA- 2006/06/23 09:00
CRDT- 2006/04/25 09:00
PHST- 2006/02/19 00:00 [received]
PHST- 2006/02/21 00:00 [accepted]
PHST- 2006/04/25 09:00 [pubmed]
PHST- 2006/06/23 09:00 [medline]
PHST- 2006/04/25 09:00 [entrez]
AID - S0006-291X(06)00424-4 [pii]
AID - 10.1016/j.bbrc.2006.02.173 [doi]
PST - ppublish
SO  - Biochem Biophys Res Commun. 2006 Jun 2;344(2):571-80. doi: 
      10.1016/j.bbrc.2006.02.173. Epub 2006 Mar 10.