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Torsin A Localization in the Mouse Cerebellar Synaptic Circuitry.

Puglisi F, Vanni V, Ponterio G, Tassone A, Sciamanna G, Bonsi P, Pisani A, Mandolesi G - PLoS ONE (2013)

Bottom Line: Torsin A (TA) is a ubiquitous protein belonging to the superfamily of proteins called "ATPases associated with a variety of cellular activities" (AAA(+) ATPase).In addition, abundant expression of the protein was found in the main GABA-ergic and glutamatergic inputs of the cerebellar cortex.These results extend our knowledge on TA synaptic localization providing a clue to its potential role in synaptic development.

View Article: PubMed Central - PubMed

Affiliation: Department of Systems Medicine, University of Rome Tor Vergata/Laboratory of Neurophysiology and Synaptic Plasticity, Fondazione Santa Lucia, Rome, Italy.

ABSTRACT
Torsin A (TA) is a ubiquitous protein belonging to the superfamily of proteins called "ATPases associated with a variety of cellular activities" (AAA(+) ATPase). To date, a great deal of attention has been focused on neuronal TA since its mutant form causes early-onset (DYT1) torsion dystonia, an inherited movement disorder characterized by sustained muscle contractions and abnormal postures. Interestingly, it has been proposed that TA, by interacting with the cytoskeletal network, may contribute to the control of neurite outgrowth and/or by acting as a chaperone at synapses could affect synaptic vesicle turnover and neurotransmitter release. Accordingly, both its peculiar developmental expression in striatum and cerebellum and evidence from DYT1 knock-in mice suggest that TA may influence dendritic arborization and synaptogenesis in the brain. Therefore, to better understand TA function a detailed description of its localization at synaptic level is required. Here, we characterized by means of rigorous quantitative confocal analysis TA distribution in the mouse cerebellum at postnatal day 14 (P14), when both cerebellar synaptogenesis and TA expression peak. We observed that the protein is broadly distributed both in cerebellar cortex and in the deep cerebellar nuclei (DCN). Of note, Purkinje cells (PC) express high levels of TA also in the spines and axonal terminals. In addition, abundant expression of the protein was found in the main GABA-ergic and glutamatergic inputs of the cerebellar cortex. Finally, TA was observed also in glial cells, a cellular population little explored so far. These results extend our knowledge on TA synaptic localization providing a clue to its potential role in synaptic development.

No MeSH data available.


Related in: MedlinePlus

TA localization in the mouse cerebellar cortex at P14.A-A’’and B–B’’) Cerebellar sagittal section immunostained for PV (green) and TA (red) and counterstained with Dapi (cyan). A) Merge image between Dapi-positive nuclei (cyan) and PV-positive PC dendrites (green) in the upper part of the ML and the EGL. A’) Shows the corresponding TA immunostaining (red). A’’) Merge image of all markers (PV-TA-Dapi) plus the colocalization mask TA/PV (mTA/PV in white) highlighting the expression of TA in PC dendrites and spines. B) Merge image of PV staining and Dapi-positive nuclei, highlighting the PV-positive PC bodies and dendrites and interneurons of the ML (green). The inset (white box) is an high magnification of a BC body. B’) TA protein (red) is abundantly present in PCs and BCs (inset), both in the cytoplasm and nuclei. Black spaces are TA-negative nuclei in the ML. B’’) merge image of all markers (PV-TA-Dapi) plus the colocalization mask TA/PV (mTA/PV in white). C) Merge image between DAPI-positive nuclei and V1-positive (green) rosettes to define the IGL. Most of the DAPI-positive nuclei in the IGL belong to granule cells C’) TA staining is diffuse also in this layer; granule cells (high magnification in the inset) and Golgi-like cells are intensively labeled. C’’) Merge image of all markers (TA-V1-Dapi) plus the colocalization mask TA/Dapi (mTA/Dapi in white). The absence of the white mask highlights no expression of TA in almost all cell nuclei. Few white dots in the inset indicate overlapping signal of TA around the nucleus. Scale bars in A, B and C 10 µm; in the insets 2 µm.
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pone-0068063-g002: TA localization in the mouse cerebellar cortex at P14.A-A’’and B–B’’) Cerebellar sagittal section immunostained for PV (green) and TA (red) and counterstained with Dapi (cyan). A) Merge image between Dapi-positive nuclei (cyan) and PV-positive PC dendrites (green) in the upper part of the ML and the EGL. A’) Shows the corresponding TA immunostaining (red). A’’) Merge image of all markers (PV-TA-Dapi) plus the colocalization mask TA/PV (mTA/PV in white) highlighting the expression of TA in PC dendrites and spines. B) Merge image of PV staining and Dapi-positive nuclei, highlighting the PV-positive PC bodies and dendrites and interneurons of the ML (green). The inset (white box) is an high magnification of a BC body. B’) TA protein (red) is abundantly present in PCs and BCs (inset), both in the cytoplasm and nuclei. Black spaces are TA-negative nuclei in the ML. B’’) merge image of all markers (PV-TA-Dapi) plus the colocalization mask TA/PV (mTA/PV in white). C) Merge image between DAPI-positive nuclei and V1-positive (green) rosettes to define the IGL. Most of the DAPI-positive nuclei in the IGL belong to granule cells C’) TA staining is diffuse also in this layer; granule cells (high magnification in the inset) and Golgi-like cells are intensively labeled. C’’) Merge image of all markers (TA-V1-Dapi) plus the colocalization mask TA/Dapi (mTA/Dapi in white). The absence of the white mask highlights no expression of TA in almost all cell nuclei. Few white dots in the inset indicate overlapping signal of TA around the nucleus. Scale bars in A, B and C 10 µm; in the insets 2 µm.

Mentions: The four layers of the juvenile cerebellar cortex (ML, PCL, EGL and IGL) were easily recognized in sagittal cerebellar sections by using the nuclear marker (Dapi) which labels almost all the cellular nuclei (Figures 2 and 3). In addition, to better define the expression of TA in different cell types, we used specific markers in combination with topological and morphological characteristics. Finally, we report the colocalization signal (mask) between TA and the specific marker to highlight the expression of the protein in the different neuronal populations of the cerebellar cortex.


Torsin A Localization in the Mouse Cerebellar Synaptic Circuitry.

Puglisi F, Vanni V, Ponterio G, Tassone A, Sciamanna G, Bonsi P, Pisani A, Mandolesi G - PLoS ONE (2013)

TA localization in the mouse cerebellar cortex at P14.A-A’’and B–B’’) Cerebellar sagittal section immunostained for PV (green) and TA (red) and counterstained with Dapi (cyan). A) Merge image between Dapi-positive nuclei (cyan) and PV-positive PC dendrites (green) in the upper part of the ML and the EGL. A’) Shows the corresponding TA immunostaining (red). A’’) Merge image of all markers (PV-TA-Dapi) plus the colocalization mask TA/PV (mTA/PV in white) highlighting the expression of TA in PC dendrites and spines. B) Merge image of PV staining and Dapi-positive nuclei, highlighting the PV-positive PC bodies and dendrites and interneurons of the ML (green). The inset (white box) is an high magnification of a BC body. B’) TA protein (red) is abundantly present in PCs and BCs (inset), both in the cytoplasm and nuclei. Black spaces are TA-negative nuclei in the ML. B’’) merge image of all markers (PV-TA-Dapi) plus the colocalization mask TA/PV (mTA/PV in white). C) Merge image between DAPI-positive nuclei and V1-positive (green) rosettes to define the IGL. Most of the DAPI-positive nuclei in the IGL belong to granule cells C’) TA staining is diffuse also in this layer; granule cells (high magnification in the inset) and Golgi-like cells are intensively labeled. C’’) Merge image of all markers (TA-V1-Dapi) plus the colocalization mask TA/Dapi (mTA/Dapi in white). The absence of the white mask highlights no expression of TA in almost all cell nuclei. Few white dots in the inset indicate overlapping signal of TA around the nucleus. Scale bars in A, B and C 10 µm; in the insets 2 µm.
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pone-0068063-g002: TA localization in the mouse cerebellar cortex at P14.A-A’’and B–B’’) Cerebellar sagittal section immunostained for PV (green) and TA (red) and counterstained with Dapi (cyan). A) Merge image between Dapi-positive nuclei (cyan) and PV-positive PC dendrites (green) in the upper part of the ML and the EGL. A’) Shows the corresponding TA immunostaining (red). A’’) Merge image of all markers (PV-TA-Dapi) plus the colocalization mask TA/PV (mTA/PV in white) highlighting the expression of TA in PC dendrites and spines. B) Merge image of PV staining and Dapi-positive nuclei, highlighting the PV-positive PC bodies and dendrites and interneurons of the ML (green). The inset (white box) is an high magnification of a BC body. B’) TA protein (red) is abundantly present in PCs and BCs (inset), both in the cytoplasm and nuclei. Black spaces are TA-negative nuclei in the ML. B’’) merge image of all markers (PV-TA-Dapi) plus the colocalization mask TA/PV (mTA/PV in white). C) Merge image between DAPI-positive nuclei and V1-positive (green) rosettes to define the IGL. Most of the DAPI-positive nuclei in the IGL belong to granule cells C’) TA staining is diffuse also in this layer; granule cells (high magnification in the inset) and Golgi-like cells are intensively labeled. C’’) Merge image of all markers (TA-V1-Dapi) plus the colocalization mask TA/Dapi (mTA/Dapi in white). The absence of the white mask highlights no expression of TA in almost all cell nuclei. Few white dots in the inset indicate overlapping signal of TA around the nucleus. Scale bars in A, B and C 10 µm; in the insets 2 µm.
Mentions: The four layers of the juvenile cerebellar cortex (ML, PCL, EGL and IGL) were easily recognized in sagittal cerebellar sections by using the nuclear marker (Dapi) which labels almost all the cellular nuclei (Figures 2 and 3). In addition, to better define the expression of TA in different cell types, we used specific markers in combination with topological and morphological characteristics. Finally, we report the colocalization signal (mask) between TA and the specific marker to highlight the expression of the protein in the different neuronal populations of the cerebellar cortex.

Bottom Line: Torsin A (TA) is a ubiquitous protein belonging to the superfamily of proteins called "ATPases associated with a variety of cellular activities" (AAA(+) ATPase).In addition, abundant expression of the protein was found in the main GABA-ergic and glutamatergic inputs of the cerebellar cortex.These results extend our knowledge on TA synaptic localization providing a clue to its potential role in synaptic development.

View Article: PubMed Central - PubMed

Affiliation: Department of Systems Medicine, University of Rome Tor Vergata/Laboratory of Neurophysiology and Synaptic Plasticity, Fondazione Santa Lucia, Rome, Italy.

ABSTRACT
Torsin A (TA) is a ubiquitous protein belonging to the superfamily of proteins called "ATPases associated with a variety of cellular activities" (AAA(+) ATPase). To date, a great deal of attention has been focused on neuronal TA since its mutant form causes early-onset (DYT1) torsion dystonia, an inherited movement disorder characterized by sustained muscle contractions and abnormal postures. Interestingly, it has been proposed that TA, by interacting with the cytoskeletal network, may contribute to the control of neurite outgrowth and/or by acting as a chaperone at synapses could affect synaptic vesicle turnover and neurotransmitter release. Accordingly, both its peculiar developmental expression in striatum and cerebellum and evidence from DYT1 knock-in mice suggest that TA may influence dendritic arborization and synaptogenesis in the brain. Therefore, to better understand TA function a detailed description of its localization at synaptic level is required. Here, we characterized by means of rigorous quantitative confocal analysis TA distribution in the mouse cerebellum at postnatal day 14 (P14), when both cerebellar synaptogenesis and TA expression peak. We observed that the protein is broadly distributed both in cerebellar cortex and in the deep cerebellar nuclei (DCN). Of note, Purkinje cells (PC) express high levels of TA also in the spines and axonal terminals. In addition, abundant expression of the protein was found in the main GABA-ergic and glutamatergic inputs of the cerebellar cortex. Finally, TA was observed also in glial cells, a cellular population little explored so far. These results extend our knowledge on TA synaptic localization providing a clue to its potential role in synaptic development.

No MeSH data available.


Related in: MedlinePlus