scholarly journals Adult medial habenula neurons require GDNF receptor GFRα1 for synaptic stability and function

PLoS Biology ◽  
2021 ◽  
Vol 19 (11) ◽  
pp. e3001350
Author(s):  
Diana Fernández-Suárez ◽  
Favio A. Krapacher ◽  
Katarzyna Pietrajtis ◽  
Annika Andersson ◽  
Lilian Kisiswa ◽  
...  
Keyword(s):  
Synaptic Plasticity ◽  
Proximity Ligation Assay ◽  
Glutamatergic Synapses ◽  
Proximity Ligation ◽  
Postsynaptic Currents ◽  
Adult Mice ◽  
Medial Habenula ◽  
Factor Receptor Alpha ◽  
And Function ◽  
Anxiety And Fear

The medial habenula (mHb) is an understudied small brain nucleus linking forebrain and midbrain structures controlling anxiety and fear behaviors. The mechanisms that maintain the structural and functional integrity of mHb neurons and their synapses remain unknown. Using spatiotemporally controlled Cre-mediated recombination in adult mice, we found that the glial cell–derived neurotrophic factor receptor alpha 1 (GFRα1) is required in adult mHb neurons for synaptic stability and function. mHb neurons express some of the highest levels of GFRα1 in the mouse brain, and acute ablation of GFRα1 results in loss of septohabenular and habenulointerpeduncular glutamatergic synapses, with the remaining synapses displaying reduced numbers of presynaptic vesicles. Chemo- and optogenetic studies in mice lacking GFRα1 revealed impaired circuit connectivity, reduced AMPA receptor postsynaptic currents, and abnormally low rectification index (R.I.) of AMPARs, suggesting reduced Ca2+ permeability. Further biochemical and proximity ligation assay (PLA) studies defined the presence of GluA1/GluA2 (Ca2+ impermeable) as well as GluA1/GluA4 (Ca2+ permeable) AMPAR complexes in mHb neurons, as well as clear differences in the levels and association of AMPAR subunits with mHb neurons lacking GFRα1. Finally, acute loss of GFRα1 in adult mHb neurons reduced anxiety-like behavior and potentiated context-based fear responses, phenocopying the effects of lesions to septal projections to the mHb. These results uncover an unexpected function for GFRα1 in the maintenance and function of adult glutamatergic synapses and reveal a potential new mechanism for regulating synaptic plasticity in the septohabenulointerpeduncular pathway and attuning of anxiety and fear behaviors.

scholarly journals Adult medial habenula neurons require GDNF receptor GFRalpha1 for synaptic stability and function

2021 ◽  
Author(s):  
Diana Fernandez-Suarez ◽  
Favio Krapacher ◽  
Katarzyna Pietrajtis ◽  
Annika Andersson ◽  
Lilian Kisiswa ◽  
...  
Keyword(s):  
Synaptic Plasticity ◽  
Proximity Ligation Assay ◽  
Glutamatergic Synapses ◽  
Genetic Studies ◽  
Proximity Ligation ◽  
Postsynaptic Currents ◽  
Adult Mice ◽  
Medial Habenula ◽  
And Function ◽  
Anxiety And Fear

The medial habenula (mHb) is an understudied small brain nucleus linking forebrain and midbrain structures controlling anxiety and fear behaviors. The mechanisms that maintain the structural and functional integrity of mHb neurons and their synapses remain unknown. Using spatio-temporally controlled Cre-mediated recombination in adult mice, we found that the GDNF receptor alpha 1 (GFRα1) is required in adult mHb neurons for synaptic stability and function. mHb neurons express some of the highest levels of GFRα1 in the mouse brain, and acute ablation of GFRα1 results in loss of septo-habenular and habenulo-interpeduncular glutamatergic synapses, with the remaining synapses displaying reduced numbers of presynaptic vesicles. Chemo- and opto-genetic studies in mice lacking GFRα1 revealed impaired circuit connectivity, reduced AMPA receptor postsynaptic currents, and abnormally low rectification index of AMPARs, suggesting reduced Ca 2+ -permeability. Further biochemical and proximity ligation assay studies defined the presence of GluA1/GluA2 (Ca 2+ -impermeable) as well as  GluA1/GluA4 (Ca 2+ -permeable) AMPAR complexes in mHb neurons, as well as clear differences in the levels and association of AMPAR subunits in mHb neurons lacking GFRα1. Finally, acute loss of GFRα1 in adult mHb neurons reduced anxiety-like behavior and potentiated context-based fear responses, phenocopying the effects of lesions to septal projections to the mHb. These results uncover an unexpected function for GFRα1 in the maintenance and function of adult glutamatergic synapses, and reveal a potential new mechanism for regulating synaptic plasticity in the septo-habenulo-interpeduncular pathway and attuning of anxiety and fear behaviors.

scholarly journals SYNPLA: A synapse-specific method for identifying learning-induced synaptic plasticity loci

2018 ◽  
Author(s):  
Kim Dore ◽  
Yvonne Pao ◽  
Jose Soria Lopez ◽  
Sage Aronson ◽  
Huiqing Zhan ◽  
...  
Keyword(s):  
Synaptic Plasticity ◽  
High Throughput ◽  
Neural Circuits ◽  
Synaptic Strength ◽  
Proximity Ligation Assay ◽  
Specific Method ◽  
Neural Pathways ◽  
Proximity Ligation ◽  
Synaptic Changes ◽  
Specific Learning

AbstractWhich neural circuits undergo synaptic changes when an animal learns? Although it is widely accepted that changes in synaptic strength underlie many forms of learning and memory, it remains challenging to connect changes in synaptic strength at specific neural pathways to specific behaviors and memories. Here we introduce SYNPLA (SYNaptic Proximity Ligation Assay), a synapse-specific, high-throughput and potentially brain-wide method capable of detecting circuit-specific learning-induced synaptic plasticity.

scholarly journals SYNPLA, a method to identify synapses displaying plasticity after learning

2020 ◽  
Vol 117 (6) ◽  
pp. 3214-3219 ◽  
Author(s):  
Kim Dore ◽  
Yvonne Pao ◽  
Jose Soria Lopez ◽  
Sage Aronson ◽  
Huiqing Zhan ◽  
...  
Keyword(s):  
Synaptic Plasticity ◽  
Learning And Memory ◽  
High Throughput ◽  
Neural Circuits ◽  
Synaptic Strength ◽  
Proximity Ligation Assay ◽  
Neural Pathways ◽  
Proximity Ligation ◽  
Synaptic Changes ◽  
Specific Learning

Which neural circuits undergo synaptic changes when an animal learns? Although it is widely accepted that changes in synaptic strength underlie many forms of learning and memory, it remains challenging to connect changes in synaptic strength at specific neural pathways to specific behaviors and memories. Here we introduce SYNPLA (synaptic proximity ligation assay), a synapse-specific, high-throughput, and potentially brain-wide method capable of detecting circuit-specific learning-induced synaptic plasticity.

scholarly journals Environmental enrichment enhances synaptic plasticity by internalization of striatal dopamine transporters

2016 ◽  
Vol 36 (12) ◽  
pp. 2122-2133 ◽  
Author(s):  
Myung-Sun Kim ◽  
Ji Hea Yu ◽  
Chul Hoon Kim ◽  
Jae Yong Choi ◽  
Jung Hwa Seo ◽  
...  
Keyword(s):  
Synaptic Plasticity ◽  
Environmental Enrichment ◽  
Functional Improvement ◽  
Proximity Ligation Assay ◽  
Proximity Ligation ◽  
Treatment Benefits ◽  
Positron Emission ◽  
Walking Tests ◽  
Relationship Of

Environmental enrichment (EE) with a complex combination of physical, cognitive and social stimulations enhances synaptic plasticity and behavioral function. However, the mechanism remains to be elucidated in detail. We aimed to investigate dopamine-related synaptic plasticity underlying functional improvement after EE. For this, six-week-old CD-1 mice were randomly allocated to EE or standard conditions for two months. EE significantly enhanced behavioral functions such as rotarod and ladder walking tests. In a [18F]FPCIT positron emission tomography scan, binding values of striatal DAT were significantly decreased approximately 18% in the EE mice relative to the control mice. DAT inhibitor administrated to establish the relationship of the DAT down-regulation to the treatment effects also improved rotarod performances, suggesting that DAT inhibition recapitulated EE-mediated treatment benefits. Next, EE-induced internalization of DAT was confirmed using a surface biotinylation assay. In situ proximity ligation assay and immunoprecipitation demonstrated that EE significantly increased the phosphorylation of striatal DAT as well as the levels of DAT bound with protein kinase C (PKC). In conclusion, we suggest that EE enables phosphorylation of striatal DAT via a PKC-mediated pathway and causes DAT internalization. This is the first report to suggest an EE-mediated mechanism of synaptic plasticity by internalization of striatal DAT.

Proximity Ligation Assay (PLA) Protocol Using Duolink® for T Cells

BIO-PROTOCOL ◽  
2016 ◽  
Vol 6 (10) ◽  
Author(s):  
Valentin Derangère ◽  
Mélanie Bruchard ◽  
Frédérique Végran ◽  
François Ghiringhelli
Keyword(s):  
T Cells ◽  
Proximity Ligation Assay ◽  
Proximity Ligation

scholarly journals Mice with cleavage-resistant N-cadherin exhibit synapse anomaly in the hippocampus and outperformance in spatial learning tasks

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
M. Asada-Utsugi ◽  
K. Uemura ◽  
M. Kubota ◽  
Y. Noda ◽  
Y. Tashiro ◽  
...  
Keyword(s):  
Memory Function ◽  
Three Dimensional ◽  
Excitatory Synapses ◽  
Missense Mutations ◽  
Glutamatergic Synapses ◽  
Learning Tasks ◽  
Transmission Electron ◽  
Nmdar Activation ◽  
And Function

AbstractN-cadherin is a homophilic cell adhesion molecule that stabilizes excitatory synapses, by connecting pre- and post-synaptic termini. Upon NMDA receptor (NMDAR) activation by glutamate, membrane-proximal domains of N-cadherin are cleaved serially by a-disintegrin-and-metalloprotease 10 (ADAM10) and then presenilin 1(PS1, catalytic subunit of the γ-secretase complex). To assess the physiological significance of the initial N-cadherin cleavage, we engineer the mouse genome to create a knock-in allele with tandem missense mutations in the mouse N-cadherin/Cadherin-2 gene (Cdh2R714G, I715D, or GD) that confers resistance on proteolysis by ADAM10 (GD mice). GD mice showed a better performance in the radial maze test, with significantly less revisiting errors after intervals of 30 and 300 s than WT, and a tendency for enhanced freezing in fear conditioning. Interestingly, GD mice reveal higher complexity in the tufts of thorny excrescence in the CA3 region of the hippocampus. Fine morphometry with serial section transmission electron microscopy (ssTEM) and three-dimensional (3D) reconstruction reveals significantly higher synaptic density, significantly smaller PSD area, and normal dendritic spine volume in GD mice. This knock-in mouse has provided in vivo evidence that ADAM10-mediated cleavage is a critical step in N-cadherin shedding and degradation and involved in the structure and function of glutamatergic synapses, which affect the memory function.

scholarly journals PDGFRα: Expression and Function during Mitral Valve Morphogenesis

2021 ◽  
Vol 8 (3) ◽  
pp. 28
Author(s):  
Kelsey Moore ◽  
Diana Fulmer ◽  
Lilong Guo ◽  
Natalie Koren ◽  
Janiece Glover ◽  
...  
Keyword(s):  
Mitral Valve ◽  
Primary Cilia ◽  
Growth Factor Receptor ◽  
Akt Phosphorylation ◽  
Valve Development ◽  
Biochemical Assays ◽  
Factor Receptor Alpha ◽  
Mesenchymal Transformation ◽  
And Function

Mitral valve prolapse (MVP) is a common form of valve disease and can lead to serious secondary complications. The recent identification of MVP causal mutations in primary cilia-related genes has prompted the investigation of cilia-mediated mechanisms of disease inception. Here, we investigate the role of platelet-derived growth factor receptor-alpha (PDGFRα), a receptor known to be present on the primary cilium, during valve development using genetically modified mice, biochemical assays, and high-resolution microscopy. While PDGFRα is expressed throughout the ciliated valve interstitium early in development, its expression becomes restricted on the valve endocardium by birth and through adulthood. Conditional ablation of Pdgfra with Nfatc1-enhancer Cre led to significantly enlarged and hypercellular anterior leaflets with disrupted endothelial adhesions, activated ERK1/2, and a dysregulated extracellular matrix. In vitro culture experiments confirmed a role in suppressing ERK1/2 activation while promoting AKT phosphorylation. These data suggest that PDGFRα functions to suppress mesenchymal transformation and disease phenotypes by stabilizing the valve endocardium through an AKT/ERK pathway.

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