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Vol. 55, Issue 5, 804-811, May 1999
Howard Hughes Medical Institute, Department of
Biochemistry, New York University Medical Center, New York, New York
(F.S.V., B.S., E.Z.);
Department of Anatomy (A.A.A.) and
Department of
Pharmacology and Toxicology (V.K.K., E.K.), Institute of Biomedicine,
University of Helsinki, Helsinki, Finland; and
Department of Biology,
Abo Akademi University, Biocity, Turku, Finland (M-L.N., M.L.,
K.K., P.P.)
Neuropeptides FF (NPFF), AF (NPAF), and SF (NPSF) are homologous
amidated peptides that were originally identified on the basis of
similarity to the molluscan neuropeptide FMRF-amide. They have been
hypothesized to have wide-ranging functions in the mammalian central
nervous system, including pain modulation, opiate function,
cardiovascular regulation, and neuroendocrine function. We have cloned
the NPFF gene from human, bovine, rat, and mouse, and show that the
precursor mRNA encodes for all three of the biochemically identified
peptides (NPFF, NPAF, and NPSF). We demonstrate that NPFF precursor
mRNA expression by Northern analysis and map sites of expression by in
situ hybridization. We confirm the validity of the in situ
hybridization by showing that its distribution in the brain and spinal
cord matches the distribution of NPFF and NPSF immunoreactivity. We go
on to show that the mRNA levels (as measured by in situ hybridization)
in the spinal cord can be up-regulated by a model for inflammatory pain
(carrageenan injection), but not by a model for neuropathic pain
(lumbar nerve ligation). Our results confirm the evolutionary conservation of NPFF, NPAF, and NPSF neuropeptide expression in mammalian brain. They also provide a context for the interpretation of
the pain-sensitizing effects of injections of these peptides that have
been previously reported. Our results support a model for the role of
these peptides in pain regulation at the level of the spinal cord.
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