dc.creatorBlaustein Kappelmacher, Matias
dc.creatorWirth, Sonia Alejandra
dc.creatorSaldaña, G.
dc.creatorPiantanida, Ana Paula
dc.creatorBogetti, Maria Eugenia
dc.creatorMartin, Maria Eugenia
dc.creatorColman Lerner, Alejandro Ariel
dc.creatorUchitel, Osvaldo Daniel
dc.date.accessioned2021-10-08T01:37:28Z
dc.date.accessioned2022-10-15T10:50:15Z
dc.date.available2021-10-08T01:37:28Z
dc.date.available2022-10-15T10:50:15Z
dc.date.created2021-10-08T01:37:28Z
dc.date.issued2020-11-24
dc.identifierBlaustein Kappelmacher, Matias; Wirth, Sonia Alejandra; Saldaña, G.; Piantanida, Ana Paula; Bogetti, Maria Eugenia; et al.; A new tool to sense pH changes at the neuromuscular junction synaptic cleft; Nature; Scientific Reports; 10; 20480; 24-11-2020; 1-7
dc.identifier2045-2322
dc.identifierhttp://hdl.handle.net/11336/143213
dc.identifierCONICET Digital
dc.identifierCONICET
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/4377544
dc.description.abstractSynaptic transmission triggers transient acidification of the synaptic cleft. Recently, it has been shown that pH affects the opening of postsynaptic channels and therefore the production of tools that allow to study these behaviors should result of paramount value. We fused α-bungarotoxin, a neurotoxin derived from the snake Bungarus multicinctus that binds irreversibly to the acetylcholine receptor extracellular domain, to the pH sensitive GFP Super Ecliptic pHluorin, and efficiently expressed it in Pichia pastoris. This sensor allows synaptic changes in pH to be measured without the need of incorporating transgenes into animal cells. Here, we show that incubation of the mouse levator auris muscle with a solution containing this recombinant protein is enough to fluorescently label the endplate post synaptic membrane. Furthermore, we could physiologically alter and measure post synaptic pH by evaluating changes in the fluorescent signal of pHluorin molecules bound to acetylcholine receptors. In fact, using this tool we were able to detect a drop in 0.01 to 0.05 pH units in the vicinity of the acetylcholine receptors following vesicle exocytosis triggered by nerve electrical stimulation. Further experiments will allow to learn the precise changes in pH during and after synaptic activation.
dc.languageeng
dc.publisherNature
dc.relationinfo:eu-repo/semantics/altIdentifier/url/http://www.nature.com/articles/s41598-020-77154-3
dc.relationinfo:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1038/s41598-020-77154-3
dc.rightshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectElectrophisiology
dc.subjectneuromuscular junction
dc.subjectsinaptic cleft
dc.subjectpH
dc.titleA new tool to sense pH changes at the neuromuscular junction synaptic cleft
dc.typeinfo:eu-repo/semantics/article
dc.typeinfo:ar-repo/semantics/artículo
dc.typeinfo:eu-repo/semantics/publishedVersion


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