Spatial distribution of DARPP-32 in dendritic spines.

The phosphoprotein DARPP-32 (dopamine and cyclic adenosine 3´, 5´-monophosphate-regulated phosphoprotein, 32 kDa) is an important component in the molecular regulation of postsynaptic signaling in neostriatum. Despite the importance of this phosphoprotein, there is as yet little known about the nano...

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Autores principales:	Hans Blom, Daniel Rönnlund, Lena Scott, Linda Westin, Jerker Widengren, Anita Aperia, Hjalmar Brismar
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/bc38b27a2a09444abbe2a2964ed6fcce
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spelling	oai:doaj.org-article:bc38b27a2a09444abbe2a2964ed6fcce2021-11-18T08:55:53ZSpatial distribution of DARPP-32 in dendritic spines.1932-620310.1371/journal.pone.0075155https://doaj.org/article/bc38b27a2a09444abbe2a2964ed6fcce2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24058659/?tool=EBIhttps://doaj.org/toc/1932-6203The phosphoprotein DARPP-32 (dopamine and cyclic adenosine 3´, 5´-monophosphate-regulated phosphoprotein, 32 kDa) is an important component in the molecular regulation of postsynaptic signaling in neostriatum. Despite the importance of this phosphoprotein, there is as yet little known about the nanoscale distribution of DARPP-32. In this study we applied superresolution stimulated emission depletion microscopy (STED) to assess the expression and distribution of DARPP-32 in striatal neurons. Primary culture of striatal neurons were immunofluorescently labeled for DARPP-32 with Alexa-594 and for the dopamine D1 receptor (D1R) with atto-647N. Dual-color STED microscopy revealed discrete localizations of DARPP-32 and D1R in the spine structure, with clustered distributions in both head and neck. Dissected spine structures reveal that the DARPP-32 signal rarely overlapped with the D1R signal. The D1R receptor is positioned in an "aggregated" manner primarily in the spine head and to some extent in the neck, while DARPP-32 forms several neighboring small nanoclusters spanning the whole spine structure. The DARPP-32 clusters have a mean size of 52 +/- 6 nm, which is close to the resolution limit of the microscope and corresponds to the physical size of a few individual phosphoprotein immunocomplexes. Dissection of synaptic proteins using superresolution microscopy gives possibilities to reveal in better detail biologically relevant information, as compared to diffraction-limited microscopy. In this work, the dissected postsynaptic topology of the DARPP-32 phosphoprotein provides strong evidence for a compartmentalized and confined distribution in dendritic spines. The protein topology and the relatively low copy number of phosphoprotein provides a conception of DARPP-32's possibilities to fine-tune the regulation of synaptic signaling, which should have an impact on the performance of the neuronal circuits in which it is expressed.Hans BlomDaniel RönnlundLena ScottLinda WestinJerker WidengrenAnita AperiaHjalmar BrismarPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 9, p e75155 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hans Blom Daniel Rönnlund Lena Scott Linda Westin Jerker Widengren Anita Aperia Hjalmar Brismar Spatial distribution of DARPP-32 in dendritic spines.
description	The phosphoprotein DARPP-32 (dopamine and cyclic adenosine 3´, 5´-monophosphate-regulated phosphoprotein, 32 kDa) is an important component in the molecular regulation of postsynaptic signaling in neostriatum. Despite the importance of this phosphoprotein, there is as yet little known about the nanoscale distribution of DARPP-32. In this study we applied superresolution stimulated emission depletion microscopy (STED) to assess the expression and distribution of DARPP-32 in striatal neurons. Primary culture of striatal neurons were immunofluorescently labeled for DARPP-32 with Alexa-594 and for the dopamine D1 receptor (D1R) with atto-647N. Dual-color STED microscopy revealed discrete localizations of DARPP-32 and D1R in the spine structure, with clustered distributions in both head and neck. Dissected spine structures reveal that the DARPP-32 signal rarely overlapped with the D1R signal. The D1R receptor is positioned in an "aggregated" manner primarily in the spine head and to some extent in the neck, while DARPP-32 forms several neighboring small nanoclusters spanning the whole spine structure. The DARPP-32 clusters have a mean size of 52 +/- 6 nm, which is close to the resolution limit of the microscope and corresponds to the physical size of a few individual phosphoprotein immunocomplexes. Dissection of synaptic proteins using superresolution microscopy gives possibilities to reveal in better detail biologically relevant information, as compared to diffraction-limited microscopy. In this work, the dissected postsynaptic topology of the DARPP-32 phosphoprotein provides strong evidence for a compartmentalized and confined distribution in dendritic spines. The protein topology and the relatively low copy number of phosphoprotein provides a conception of DARPP-32's possibilities to fine-tune the regulation of synaptic signaling, which should have an impact on the performance of the neuronal circuits in which it is expressed.
format	article
author	Hans Blom Daniel Rönnlund Lena Scott Linda Westin Jerker Widengren Anita Aperia Hjalmar Brismar
author_facet	Hans Blom Daniel Rönnlund Lena Scott Linda Westin Jerker Widengren Anita Aperia Hjalmar Brismar
author_sort	Hans Blom
title	Spatial distribution of DARPP-32 in dendritic spines.
title_short	Spatial distribution of DARPP-32 in dendritic spines.
title_full	Spatial distribution of DARPP-32 in dendritic spines.
title_fullStr	Spatial distribution of DARPP-32 in dendritic spines.
title_full_unstemmed	Spatial distribution of DARPP-32 in dendritic spines.
title_sort	spatial distribution of darpp-32 in dendritic spines.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/bc38b27a2a09444abbe2a2964ed6fcce
work_keys_str_mv	AT hansblom spatialdistributionofdarpp32indendriticspines AT danielronnlund spatialdistributionofdarpp32indendriticspines AT lenascott spatialdistributionofdarpp32indendriticspines AT lindawestin spatialdistributionofdarpp32indendriticspines AT jerkerwidengren spatialdistributionofdarpp32indendriticspines AT anitaaperia spatialdistributionofdarpp32indendriticspines AT hjalmarbrismar spatialdistributionofdarpp32indendriticspines
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Spatial distribution of DARPP-32 in dendritic spines.

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