Localization of dopamine- and cAMP-regulated phosphoprotein-32 and inhibitor-1 in area 9 of Macaca mulatta prefrontal cortex

Abstract

The actions of dopamine D1 family receptors (D1R) depend upon a signal transduction cascade that modulates the phosphorylation state of important effector proteins, such as glutamate receptors and ion channels. This is accomplished both through activation of protein kinase A (PKA) and the inhibition of protein phosphatase-1 (PP1). Inhibition of PP1 occurs through PKA-mediated phosphorylation of DARPP-32 or the related protein inhibitor-1 (I-1), and the availability of DARPP-32 is essential to the functional outcome of D1R activation in the basal ganglia. While D1R activation is critical for prefrontal cortex (PFC) function, especially working memory, the functional role played by DARPP-32 or I-1 is less clear. In order to examine this more thoroughly, we have utilized immunoelectron microscopy to quantitatively determine the localization of DARPP-32 and I-1 in the neuropil of the rhesus monkey PFC. Both were distributed widely in the different components of the neuropil, but were enriched in dendritic shafts. I-1 label was more frequently identified in axon terminals than was DARPP-32, and DARPP-32 label was more frequently identified in glia than was I-1. We also quantified the extent to which these proteins were found in dendritic spines. DARPP-32 and I-1 were present in small subpopulations of dendritic spines, (4.4 and 7.7% and respectively), which were substantially smaller than observed for D1R in our previous studies (20%). Double-label experiments did not find evidence for colocalization of D1R and DARPP-32 or I-1 in spines or terminals. Thus, at the least, not all prefrontal spines which contain D1R also contain I-1 or DARPP-32, suggesting important differences in D1R signaling in the PFC compared to the striatum. Corresponding author: E. Chris Muly, Yerkes National Primate Research Center, 954 Gatewood Rd. NE, Atlanta, GA 30329, telephone (404) 727-9603, facsimile (404) 727-3278, emuly@emory.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access Author ManuscriptNeuroscience. Author manuscript; available in PMC 2011 May 5. The functional effects of activating D 1 or D 5 , the dopamine D1 family receptors (D1R), depend on a complex signal transduction pathway. The basic scheme of D1R signal transduction involves the coupling of D1R to G αs to activate adenylyl cyclase (AC) and produce cyclic AMP (cAMP) which, in turn, activates protein kinase A (PKA). PKA phosphorylates a variety of substrate proteins, and it is the altered phosphorylation of key signaling proteins that accounts for much of the effects of D1R stimulation. The actions of PK...

Immunohistochemical Localization of DARPP-32 in the Brain of Two Lungfishes: Further Assessment of Its Relationship with the Dopaminergic System

“…Moreover, the staining pattern supports the conclusion that the DARPP-32 is present in dopaminoceptive neurons, i.e., neurons that receive a dopamine input, and that it is absent from the dopaminergic neurons themselves [Ouimet et al, 1984[Ouimet et al, , 1992Schalling et al, 1990]. In particular, the protein was primarily detected in the neostriatum [Ouimet et al, 1984[Ouimet et al, , 1992Gustafson and Greengard, 1990;Ouimet and Greengard, 1990;Anderson and Reiner, 1991;Barbas et al, 1993;Greengard et al, 1998;Wang et al, 2004;Ishikawa et al, 2007;Glausier et al, 2010], and a striatal prevalence of the protein has been also confirmed in birds [Absil et al, 2001;Schnabel et al, 1997;Durstewitz et al, 1998;Reiner et al, 1998;Roberts et al, 2002;Bálint et al, 2004;Reiner et al, 2004;Metzger et al, 2006;Bálint and Csillag, 2007;Csillag et al, 2008]. In addition, studies in reptiles confirmed a pattern of distribution largely comparable among all amniotes [Smeets et al, 2001[Smeets et al, , 2003Moreno et al, 2010].…”

“…References: teleost: Robra and Thirumalai [2016]; lungfishes: González and Northcutt [2009]; present results; amphibians: López et al [2010]; O'Connell et al [2010]; reptiles: Smeets et al [2001Smeets et al [ , 2003; Moreno et al [2010]; birds: Schnabel et al [1997]; Durstewitz et al [1998]; Reiner et al [1998]; Absil et al [2001]; Roberts et al [2002]; Bálint et al [2004]; Reiner et al [2004]; Metzger et al [2006]; Bálint and Csillag [2007]; Csillag et al [2008]; mammals: Ouimet et al [1984Ouimet et al [ , 1992; Ouimet and Greengard [1990]; Gustafson and Greengard [1990]; Anderson and Reiner [1991]; Perez and Lewis [1992]; Barbas et al [1993]; Greengard et al [1998]; Wang et al [2004]; Ishikawa et al [2007]; Glausier et al [2010]; Buesa et al [2016]. …”

“…Also, the pallium of zebrafish is devoid of DARPP-32-ir cells [Robra and Thirumalai, 2016]. In contrast, the cortical areas of reptiles, birds, and mammals contain DARPP-32-ir cells [Ouimet et al, 1984[Ouimet et al, , 1992Berger et al, 1990;Durstewitz et al, 1998;Absil et al, 2001;Smeets et al, 2001Smeets et al, , 2003Wang et al, 2004;Ishikawa et al, 2007;Csillag et al, 2008;Glausier et al, 2010], representing a constant feature shared by all amniotes studied. Particularly relevant is the presence of 304 DARPP-32-ir cells in the prefrontal cortex of mammals, which receives a strong dopaminergic innervation from the ventral tegmental area, and plays a critical role in working memory and selective attention tasks [Goldman-Rakic et al, 2000].…”

Synaptic NMDA receptor stimulation activates PP1 by inhibiting its phosphorylation by Cdk5

“…Expression of PP1 (T320A) substantially I-1 a strong candidate for inhibiting PP1 activity in those tissues. However, I-1 is not expressed in high levels in hippocampus where it is present in the dentate gyrus, albeit at low levels (Allen et al, 2000), and it was shown, by immuno-EM studies along with DARPP-32, not to be preferentially localized in synaptic spines (Glausier et al, 2010). We found that in cortical neurons only a very small fraction of I-1 was phosphorylated at T35, the form of I-1 that can inhibit PP1 potently.…”

Immunohistochemical localization of DARPP-32 in the brain and spinal cord of anuran amphibians and its relation with the catecholaminergic system

“…Previous immunohistochemical studies have analyzed the distribution of DARPP-32 in the brain of mammals (Gustafson and Greengard, 1990;Ouimet et al, 1984Ouimet et al, , 1992Ouimet and Greengard, 1990;Anderson and Reiner, 1991;Barbas et al, 1993;Greengard et al, 1998;Wang et al, 2004;Ishikawa et al, 2007;Glausier et al, 2010), birds (Schnabel et al, 1997;Durstewitz et al, 1998;Reiner et al, 1998;Absil et al, 2001;Roberts et al, 2002;Bá lint et al, 2004;Reiner et al, 2004;Metzger et al, 2006;Bá lint and Csillag, 2007;Csillag et al, 2008) and reptiles (Smeets et al, , 2003. Many of these studies were mainly focused on the basal ganglia and only a few works have analyzed the distribution of DARPP-32 in regions outside the telencephalic basal ganglia and their related mesencephalic structures (Ouimet et al, 1984(Ouimet et al, , 1992Schalling et al, 1990;Perez and Lewis, 1992;Smeets et al, 2001Smeets et al, , 2003Metzger et al, 2006).…”