ArticleJournal of visualized experiments : JoVE2015
Detection of in situ protein-protein complexes at the Drosophila larval neuromuscular junction using proximity ligation assay.
Article in Journal of visualized experiments : JoVE, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- γ-secretase promotes Drosophila postsynaptic development through the cleavage of a Wnt receptor.Developmental cell · 2022Article
- WAKE-mediated modulation of cVA perception via a hierarchical neuro-endocrine axis in Drosophila male-male courtship behaviour.Nature communications · 2022Article
- Animal Models of Neurodegenerative Disease: Recent Advances in Fly Highlight Innovative Approaches to Drug Discovery.Frontiers in molecular neuroscience · 2022Review
- Abnormal accumulation of lipid droplets in neurons induces the conversion of alpha-Synuclein to proteolytic resistant forms in a Drosophila model of Parkinson's disease.PLoS genetics · 2021Article
- βPS-Integrin acts downstream of Innexin 2 in modulating stretched cell morphogenesis in the Drosophila ovary.G3 (Bethesda, Md.) · 2021Article
- PKC Delta Activation Promotes Endoplasmic Reticulum Stress (ERS) and NLR Family Pyrin Domain-Containing 3 (NLRP3) Inflammasome Activation Subsequent to Asynuclein-Induced Microglial Activation: Involvement of Thioredoxin-Interacting Protein (TXNIP)/Thioredoxin (Trx) Redoxisome Pathway.Frontiers in aging neuroscience · 2021Article
- In Situ Detection of Ribonucleoprotein Complex Assembly in the C. elegans Germline using Proximity Ligation Assay.Journal of visualized experiments : JoVE · 2020Article
- Basigin Associates with Integrin in Order to Regulate Perineurial Glia andThe Journal of neuroscience : the official journal of the Society for Neuroscience · 2020Article
- Homeodomain-interacting protein kinase (Hipk) plays roles in nervous system and muscle structure and function.PloS one · 2020Article
- The calcium channel subunit αNature communications · 2019Article
- Complexin cooperates with Bruchpilot to tether synaptic vesicles to the active zone cytomatrix.The Journal of cell biology · 2019Article
- Levels of Par-1 kinase determine the localization of Bruchpilot at the Drosophila neuromuscular junction synapses.Scientific reports · 2018Article
- Tenectin recruits integrin to stabilize bouton architecture and regulate vesicle release at theeLife · 2018Article
- Nitric oxide-mediated posttranslational modifications control neurotransmitter release by modulating complexin farnesylation and enhancing its clamping ability.PLoS biology · 2018Article
- Article
- DISC1 causes associative memory and neurodevelopmental defects in fruit flies.Molecular psychiatry · 2016Article
- Adducin at the Neuromuscular Junction in Amyotrophic Lateral Sclerosis: Hanging on for Dear Life.Frontiers in cellular neuroscience · 2016Review
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Authors and funding
8 authors.
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Abstract
Discs large (Dlg) is a conserved member of the membrane-associated guanylate kinase family, and serves as a major scaffolding protein at the larval neuromuscular junction (NMJ) in Drosophila. Previous studies have shown that the postsynaptic distribution of Dlg at the larval NMJ overlaps with that of Hu-li tai shao (Hts), a homologue to the mammalian adducins. In addition, Dlg and Hts are observed to form a complex with each other based on co-immunoprecipitation experiments involving whole adult fly lysates. Due to the nature of these experiments, however, it was unknown whether this complex exists specifically at the NMJ during larval development. Proximity Ligation Assay (PLA) is a recently developed technique used mostly in cell and tissue culture that can detect protein-protein interactions in situ. In this assay, samples are incubated with primary antibodies against the two proteins of interest using standard immunohistochemical procedures. The primary antibodies are then detected with a specially designed pair of oligonucleotide-conjugated secondary antibodies, termed PLA probes, which can be used to generate a signal only when the two probes have bound in close proximity to each other. Thus, proteins that are in a complex can be visualized. Here, it is demonstrated how PLA can be used to detect in situ protein-protein interactions at the Drosophila larval NMJ. The technique is performed on larval body wall muscle preparations to show that a complex between Dlg and Hts does indeed exist at the postsynaptic region of NMJs.
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