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Phosphorylation of DARPP-32 and protein phosphatase inhibitor-1 in rat choroid plexus: regulation by factors other than dopamine

GL Snyder, JA Girault, JY Chen, AJ Czernik, JW Kebabian, JA Nathanson and P Greengard
Journal of Neuroscience 1 August 1992, 12 (8) 3071-3083; https://doi.org/10.1523/JNEUROSCI.12-08-03071.1992
GL Snyder
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021.
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JA Girault
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021.
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JY Chen
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021.
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AJ Czernik
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021.
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JW Kebabian
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021.
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JA Nathanson
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021.
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P Greengard
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021.
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Abstract

The molecular mechanisms underlying regulation of fluid production by secretory epithelia such as the choroid plexus are poorly understood. Two cAMP-regulated inhibitors of protein phosphatase-1, inhibitor-1 (I1) and a dopamine- and cAMP-regulated phosphoprotein, M(r) = 32,000 (DARPP-32), are enriched in the choroid plexus. We show here that these two phosphoproteins are colocalized in choroid plexus epithelial cells. We have developed a novel method for studying the phosphorylation state of DARPP-32 and I1 in intact cells, using a phosphorylation state- specific monoclonal antibody. Several drugs and hormones that are known to alter fluid secretion and that increase cAMP levels (forskolin, isoproterenol, vasoactive intestinal peptide) or cGMP levels (atrial natriuretic peptide) or that may use additional second messenger pathways (5-HT), increase the phosphorylation of I1 and DARPP-32 in rat choroid plexus. In contrast, dopamine does not alter cAMP and cGMP levels, or I1 and DARPP-32 phosphorylation. Our results indicate that DARPP-32, known to be regulated by dopamine in a number of tissues, can be phosphorylated in response to non-dopaminergic factors, including hormones acting through non-cAMP-dependent pathways. Our results also raise the possibility that inhibition of phosphatase-1, as a result of I1 and DARPP-32 phosphorylation, might be part of a final common pathway in the action of several factors that are known or thought to alter cerebrospinal fluid production.

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The Journal of Neuroscience: 12 (8)
Journal of Neuroscience
Vol. 12, Issue 8
1 Aug 1992
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Phosphorylation of DARPP-32 and protein phosphatase inhibitor-1 in rat choroid plexus: regulation by factors other than dopamine
GL Snyder, JA Girault, JY Chen, AJ Czernik, JW Kebabian, JA Nathanson, P Greengard
Journal of Neuroscience 1 August 1992, 12 (8) 3071-3083; DOI: 10.1523/JNEUROSCI.12-08-03071.1992

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Phosphorylation of DARPP-32 and protein phosphatase inhibitor-1 in rat choroid plexus: regulation by factors other than dopamine
GL Snyder, JA Girault, JY Chen, AJ Czernik, JW Kebabian, JA Nathanson, P Greengard
Journal of Neuroscience 1 August 1992, 12 (8) 3071-3083; DOI: 10.1523/JNEUROSCI.12-08-03071.1992
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