The Proximity Ligation Assay: A High Throughput Technique for Protein Analysis in Neuroscience

2015 ◽  
pp. 231-240
Author(s):  
Michela Zaltieri ◽  
PierFranco Spano ◽  
Cristina Missale ◽  
Arianna Bellucci
Keyword(s):  
High Throughput ◽  
Protein Analysis ◽  
Proximity Ligation Assay ◽  
Proximity Ligation

scholarly journals HiPLA: High-throughput imaging Proximity Ligation Assay

2018 ◽  
Author(s):  
Leonid A. Serebryannyy ◽  
Tom Misteli
Keyword(s):  
High Throughput ◽  
Protein Interactions ◽  
Large Scale ◽  
Protein Complexes ◽  
Nuclear Lamina ◽  
Cellular Protein ◽  
Premature Aging ◽  
Proximity Ligation Assay ◽  
Proximity Ligation ◽  
High Throughput Imaging

AbstractProtein-protein interactions are essential for cellular structure and function. To delineate how the intricate assembly of protein interactions contribute to cellular processes in health and disease, new methodologies that are both highly sensitive and can be applied at large scale are needed. Here, we develop HiPLA (high-throughput imaging proximity ligation assay), a method that employs the antibody-based proximity ligation assay in a high-throughput imaging screening format to systematically probe protein interactomes. Using HiPLA, we probe the interaction of 60 proteins and associated PTMs with the nuclear lamina in a model of the premature aging disorder Hutchinson-Gilford progeria syndrome (HGPS). We identify a subset of proteins that differentially interact with the nuclear lamina in HGPS. In combination with quantitative indirect immunofluorescence, we find that the majority of differential interactions were accompanied by corresponding changes in expression of the interacting protein. Taken together, HiPLA offers a novel approach to probe cellular protein-protein interaction at a large scale and reveals mechanistic insights into the assembly of protein complexes.

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 HiPLA: High-throughput imaging proximity ligation assay

Methods ◽  
2019 ◽  
Vol 157 ◽  
pp. 80-87 ◽  
Author(s):  
Leonid A. Serebryannyy ◽  
Tom Misteli
Keyword(s):  
High Throughput ◽  
Proximity Ligation Assay ◽  
Proximity Ligation ◽  
High Throughput Imaging

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 AHR:IKAROS Interaction Promotes Platelet Biogenesis in Response to SR1

Reports ◽  
2021 ◽  
Vol 4 (1) ◽  
pp. 7
Author(s):  
Lea Mallo ◽  
Valentin Do Sacramento ◽  
Christian Gachet ◽  
Susan Chan ◽  
Philippe Kastner ◽  
...  
Keyword(s):  
High Capacity ◽  
Proximity Ligation Assay ◽  
Physical Interaction ◽  
Short Report ◽  
Production Engineering ◽  
Platelet Production ◽  
Proximity Ligation ◽  
Aryl Hydrocarbon ◽  
Dependent Pathway

In vitro, the differentiation of megakaryocytes (MKs) is improved by aryl-hydrocarbon receptor (AHR) antagonists such as StemRegenin 1 (SR1), an effect physiologically recapitulated by the presence of stromal mesenchymal cells (MSC). This inhibition promotes the amplification of a CD34+CD41low population able to mature as MKs with a high capacity for platelet production. In this short report, we showed that the emergence of the thrombocytogenic precursors and the enhancement of platelet production triggered by SR1 involved IKAROS. The downregulation/inhibition of IKAROS (shRNA or lenalidomide) significantly reduced the emergence of SR1-induced thrombocytogenic population, suggesting a crosstalk between AHR and IKAROS. Interestingly, using a proximity ligation assay, we could demonstrate a physical interaction between AHR and IKAROS. This interaction was also observed in the megakaryocytic cells differentiated in the presence of MSCs. In conclusion, our study revealed a previously unknown AHR/ IKAROS -dependent pathway which prompted the expansion of the thrombocytogenic precursors. This AHR- IKAROS dependent checkpoint controlling MK maturation opens new perspectives to platelet production engineering.

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