DARPP-32 mRNA Research Articles

Regional distribution of DARPP-32 (dopamine- and adenosine 3',5'-monophosphate-regulated phosphoprotein of Mr = 32,000) mRNA in mouse brain.

DARPP-32 (dopamine- and adenosine 3',5'-monophosphate-regulated phosphoprotein of Mr = 32,000) mRNA distribution was examined in adult mouse central nervous system by in situ hybridization. In general, DARPP-32 mRNA was found in regions of brain where cells express the dopamine D1 subtype receptor. Cells of the olfactory tubercle, caudate-putamen, and nucleus accumbens had the highest levels of DARPP-32 mRNA, as did choroid plexus and Purkinje cells. Relatively high levels were found in medial habenula and lateral piriform cortex. Moderate levels were seen in cerebral cortex layer VI, medial piriform cortex, lateral entorhinal cortex, tenia tecta, anterior olfactory nucleus, and lateral bed nucleus of the stria terminalis. Low levels were observed in hippocampus, cerebral cortex layers II and III, olfactory bulb, and the nucleus of the lateral olfactory tract. DARPP-32 mRNA levels in the amygdaloid nuclei varied greatly.

Ontogeny of D 1 and DARPP-32 gene expression in the rat striatum: an in situ hybridization study

D 1 dopamine receptor (D 1R) and DARPP-32 (a dopamine and adenosine 3′,5′-monophosphate regulated phosphoprotein), gene expression was studied in the rat striatum in adults and during ontogeny by in situ hybridization. D 1R mRNA was first detected in the striatal primordium at day 17 of gestation. At day 18, D 1R mRNA was found throughout the striatum. Before birth, the striatal neurons had neuroblastic aspect and were close together, giving homogeneous and compact labelling. After birth, the topography and aspect of the neurons containing D 1R mRNA and DARPP-32 mRNA were similar. The two mRNAs were detectable in the caudate-putamen, accumbens nucleus and olfactory tubercle. The microautoradiographic analysis demonstrated that D 1R and DARPP-32 genes are massively expressed by the medium-sized striatal neurons. The proportion of medium-sized neurons containing the DARPP-32 mRNA was however higher than that of the neurons containing the D 1R mRNA. Furthermore, an unexpected proportion of large-sized neurons express these genes. This proportion varies with development. Comparison between the appearance, topography and frequency of choline-acetyltransferase immunoreactive neurons and large-sized neurons containing D 1R or DARPP-32 mRNA suggest that these large-sized neurons containing D 1R and DARPP-32 mRNAs are cholinergic ones.

DARPP-32 development in the caudate nucleus is independent of afferent input from the substantia nigra

DARPP-32, a dopamine- and adenosine 3′:5′-monophosphate regulated neuronal phosphoprotein, M r 32 kDa, is a phenotypic marker of the medium-size spiny neurons of the mammalian caudate-putamen. In the present study, we examined the ontogeny of DARPP-32 protein and mRNA, and compared it to the ontogeny of tyrosine hydroxylase and synapsin I, a synaptic-vesicle phosphoprotein. In vivo, the amount of DARPP-32 protein per mg total protein increased throughout the first three postnatal weeks, and then declined to plateau at adult levels. The mRNA level closely paralleled the protein, except that its rise preceded that of the protein. Tyrosine hydroxylase levels rose throughout the first 4 postnatal weeks, and synapsin I levels rose steadily during the same period. Primary reaggregate cultures containing cells from the caudate-putamen expressed DARPP-32 with a time course similar to that seen in vivo. The level of expression was not altered by coculturing with dopaminergic neurons from the rostral mesencephalic tegmentum. Thus, the postnatal increase in DARPP-32 levels in the caudate-putamen appears to be independent of transsynaptic or end-organ influences from the substantia nigra.

Comparison of gene expression of the dopamine D-2 receptor and DARPP-32 in rat brain, pituitary and adrenal gland

In situ hybridization histochemistry was used to compare the distribution of dopamine D-2 receptor mRNA and DARPP-32 mRNA in rat brain, pituitary and adrenal gland. In many regions such as the caudate nucleus, nucleus accumbens and olfactory tubercle, there was an excellent correlation with dopamine receptor binding studies, whereas in some discrete brain regions mismatches were observed.

Rat DARPP-32: Cloning, sequencing, and characterization of the cDNA

DARPP-32, a substrate for cyclic AMP-dependent protein kinase, is highly enriched in the caudate nucleus. In the present study, the cDNA for rat DARPP-32 was isolated and characterized. When compared to the coding region of bovine DARPP-32 cDNA, there was 86% identity at the nucleotide level, and 84% identity at the amino acid level. The homology in the region previously noted to be similar to phosphatase inhibitor-1 remained intact. There were also 3'-untranslated regions that were highly conserved. The DARPP-32 mRNA, which appeared as a doublet, was widely distributed in the central nervous system and was highly enriched in the striatum. Southern blot analysis revealed a simple hybridization pattern, consistent with the presence of a single gene coding for rat DARPP-32. Unilateral destruction of the nigrostriatal dopaminergic pathway by injection of 6-hydroxydopamine (6-OHDA) into the substantia nigra did not alter the striatal steady state levels of DARPP-32 mRNA.

Distribution and cellular localization of DARPP-32 mRNA in rat brain

In situ hybridization histochemistry has been used to determine the regional distribution and cellular localization of DARPP-32 mRNA in the rat brain. Results support the concept that DARPP-32 is present primarily in cells expressing the dopamine D 1 subtype receptor, and that DARPP-32 is not synthesized in dopamine-containing cells. Strongly labelled neuronal cell bodies were found in the caudate nucleus, nucleus accumbens, olfactory tubercle, parts of the bed nucleus of the stria terminalis, and the amygdaloid complex. In addition large amounts of DARPP-32 mRNA were visualized in the medial habenula and around the third ventricle, in ependymal cells and tanycytes, and in the cerebellar Purkinje cells. A less pronounced activity was seen in layers II–III and VI throughout the cerebral cortex. The present studies together with previous biochemical and immunocytochemical studies demonstrate that DARPP-32 gene expression is greatest primarily in D 1 dopaminoceptive cells, although there are exceptions. In situ hybridization may thus be used to quantitate regulation of DARPP-32 mRNA in discrete brain regions.

Dopamine- and cAMP-regulated phosphoprotein (DARPP-32) and dopamine DA1 agonist-sensitive Na+,K+-ATPase in renal tubule cells.

The cellular localization of DARPP-32, a dopamine- and cAMP-regulated phosphoprotein of Mr 32,000 that appears to mediate certain actions of dopamine in the mammalian brain by acting as an inhibitor of protein phosphatase 1, was studied in the kidney of several species. DARPP-32 mRNA and DARPP-32-like immunoreactivity were found in the cytoplasm of cells in the thick ascending limb of the loop of Henle. The specific dopamine DA1 agonist SKF 82526 caused a dose-dependent inhibition of Na+,K+-ATPase activity, which could be blocked by SCH 23390, a specific DA1 antagonist, and by PKI-(5-24) amide, a specific inhibitor of cAMP-dependent protein kinase. The results indicate that DA1 dopamine receptors and DARPP-32, an intracellular third messenger for dopamine, are part of the signal-transduction process for dopamine acting on renal tubule cells.