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Journal of Neuroscience, Vol 6, 1469-1481, Copyright © 1986 by Society for Neuroscience
DARPP-32, a dopamine- and adenosine 3':5'-monophosphate-regulated phosphoprotein: regional, tissue, and phylogenetic distribution
HC Hemmings Jr and P Greengard
Rabbit antisera and mouse monoclonal antibodies have been prepared to
bovine DARPP-32 (dopamine- and adenosine 3':5'-monophosphate-regulated
phosphoprotein, Mr = 32,000), and used to study its regional, tissue, and
phylogenetic distributions. The antibodies, none of which distinguished
between dephospho-DARPP-32 and phospho-DARPP-32, were characterized and
used to develop a sensitive and specific radioimmunoassay for DARPP-32. The
radioimmunoassay, in conjunction with immunolabeling of SDS/PAGE transfers
and immunoprecipitation of phosphorylated tissue extracts, was used to
measure immunoreactive DARPP-32 in microdissected regions of rat CNS, in
peripheral nervous and non-nervous tissues, and in CNS tissue from various
animal species. The distribution of DARPP-32 was generally consistent with
the interpretation that it is localized primarily to dopaminoceptive cells
that possess dopamine-sensitive adenylate cyclase (D-1 dopamine receptors
coupled to adenylate cyclase). Within the rat CNS, DARPP-32 was most highly
concentrated in the basal ganglia. DARPP-32 was present in neostriatum from
all six mammalian species tested (mouse, rat, guinea pig, rabbit, cow, and
rhesus monkey) at concentrations of from 96 to 144 pmol/mg total protein,
which constituted from 0.22 to 0.32% of the total protein. DARPP-32 was
also identified at low levels in several peripheral tissues, including
choroid plexus, parathyroid cells, adrenal chromaffin cells, posterior
pituitary gland, pineal gland, and superior cervical sympathetic ganglion.
A phylogenetic survey was carried out of proteins immunologically related
to DARPP-32 in nervous tissue from nonmammalian species. DARPP-32-like
proteins were identified in dopaminoceptive brain regions from
representative members of the amniote vertebrate classes (birds and
reptiles), while none was identified in dopaminoceptive brain regions from
representative members of the anamniote vertebrate classes (bony fishes and
amphibians) or in nervous tissue from representative members of several
invertebrate classes.
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