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Volume 271,
Number 5,
Issue of February 2, 1996 pp. 2658-2662
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
Coinduction of
Nitric Oxide Synthase, Argininosuccinate Synthetase, and
Argininosuccinate Lyase in Lipopolysaccharide-treated Rats
RNA BLOT, IMMUNOBLOT, AND IMMUNOHISTOCHEMICAL ANALYSES
(Received for publication, October 2, 1995)
Akitoshi
Nagasaki
, ,
Tomomi
Gotoh
,
Motohiro
Takeya
,
Yingjie
Yu
,
Masaki
Takiguchi
,
Hiromitsu
Matsuzaki
,
Kiyoshi
Takatsuki
,
Masataka
Mori
Nitric oxide (NO) is synthesized from arginine by nitric oxide
synthase (NOS), and citrulline which is generated can be recycled to
arginine by argininosuccinate synthetase (AS) and argininosuccinate
lyase (AL). Rats were injected with bacterial lipopolysaccharide (LPS),
and expression of the inducible isoform of NOS (iNOS), AS, and AL was
analyzed. In RNA blot analysis, iNOS mRNA was undetectable before the
LPS treatment but was induced by LPS in the lung, heart, liver, and
spleen, and less strongly in the skeletal muscle and testis. AS mRNA
was induced in the lung and spleen, and AL mRNA was weakly induced in
these tissues. AS and AL mRNAs were abundant in the control liver and
remained unchanged after the treatment. Kinetic studies showed that
iNOS mRNA increased rapidly in both spleen and lung, reached a maximum
2-5 h after the treatment, and decreased thereafter. On the other
hand, AS mRNA increased more slowly and reached a maximum in 6-12
h (by about 10-fold in the spleen and 2-fold in the lung). AL mRNA in
the spleen and lung increased slowly and remained high up to 24 h. In
immunoblot analysis, increase of iNOS protein was evident in the lung,
liver, and spleen, and there was an increase of AS protein in the lung
and spleen. In immunohistochemical analysis, macrophages in the spleen
that were negative for iNOS and AS before LPS treatment were strongly
positive for both iNOS and AS after this treatment. As iNOS, AS, and AL
were coinduced in rat tissues and cells, citrulline-arginine recycling
seems to be important in NO synthesis under the conditions of
stimulation.

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Copyright © 1996 by the American Society for Biochemistry and Molecular Biology.
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