Molecular Mechanisms Underlying ATP- And Adenosine Induced Microvascular Endothelial Barrier Preservation

Hdl Handle:
http://hdl.handle.net/10675.2/600799
Title:
Molecular Mechanisms Underlying ATP- And Adenosine Induced Microvascular Endothelial Barrier Preservation
Authors:
Batori, Robert
Abstract:
Endothelial barrier integrity has critical importance in vascular homeostasis. Disruption of the endothelial cell (EC) barrier results in increased vascular permeability. Extracellular purines, ATP and adenosine (Ado) can restore the barrier function, involving the activation of myosin light chain phosphatase (MLCP). Both ATP and Ado increase protein kinase-A (PKA) activity, however a direct link between purine-induced EC barrier enhancement, MLCP and PKA was not described. Here we show that Ado and a stable analog of ATP, ATPγS, induced human lung microvascular EC (HLMVEC) barrier enhancement and PKA activation leads to decrease in MLC and MYPT1T696 phosphorylation. Surprisingly, PKA catalytic subunit (PKAc) depletion attenuates ATPγS, but not Ado-induced increase in transendothelial electrical resistance (TER), indicating that PKA activation is involved in ATP-induced EC barrier enhancement. Depletion of PKAc leads to increase in MLC and MYPTT696 phosphorylation in ATPγS challenged EC supporting the role of PKA in MLCP activation. To elucidate the role of PKA signaling in ATP-induced EC barrier enhancement we depleted several PKA-anchoring proteins (AKAPs). AKAP2 depletion attenuates ATPγS, but not Ado-induced TER increase. Furthermore, AKAP2 co-immunoprecipitates with MYPT1. This interaction was also confirmed by PLA. In conclusion ATP- and Ado-induced barrier enhancement requires different signaling with PKA promoting ATP-, but not Ado-induced EC barrier strengthening.
Affiliation:
Vascular Biology Center
Issue Date:
Mar-2016
URI:
http://hdl.handle.net/10675.2/600799
Type:
Other
Language:
en_US
Description:
Poster presented at the 2016 Graduate Research Day
Appears in Collections:
2016 Graduate Research Day; Vascular Biology Center: Student Research and Presentations

Full metadata record

DC FieldValue Language
dc.contributor.authorBatori, Roberten
dc.date.accessioned2016-03-07T22:35:09Zen
dc.date.available2016-03-07T22:35:09Zen
dc.date.issued2016-03en
dc.identifier.urihttp://hdl.handle.net/10675.2/600799en
dc.descriptionPoster presented at the 2016 Graduate Research Dayen
dc.description.abstractEndothelial barrier integrity has critical importance in vascular homeostasis. Disruption of the endothelial cell (EC) barrier results in increased vascular permeability. Extracellular purines, ATP and adenosine (Ado) can restore the barrier function, involving the activation of myosin light chain phosphatase (MLCP). Both ATP and Ado increase protein kinase-A (PKA) activity, however a direct link between purine-induced EC barrier enhancement, MLCP and PKA was not described. Here we show that Ado and a stable analog of ATP, ATPγS, induced human lung microvascular EC (HLMVEC) barrier enhancement and PKA activation leads to decrease in MLC and MYPT1T696 phosphorylation. Surprisingly, PKA catalytic subunit (PKAc) depletion attenuates ATPγS, but not Ado-induced increase in transendothelial electrical resistance (TER), indicating that PKA activation is involved in ATP-induced EC barrier enhancement. Depletion of PKAc leads to increase in MLC and MYPTT696 phosphorylation in ATPγS challenged EC supporting the role of PKA in MLCP activation. To elucidate the role of PKA signaling in ATP-induced EC barrier enhancement we depleted several PKA-anchoring proteins (AKAPs). AKAP2 depletion attenuates ATPγS, but not Ado-induced TER increase. Furthermore, AKAP2 co-immunoprecipitates with MYPT1. This interaction was also confirmed by PLA. In conclusion ATP- and Ado-induced barrier enhancement requires different signaling with PKA promoting ATP-, but not Ado-induced EC barrier strengthening.en
dc.language.isoen_USen
dc.subjectAdenosineen
dc.subjectCatalytic Domainen
dc.subjectElectric Impedanceen
dc.subjectEndothelial Cellsen
dc.titleMolecular Mechanisms Underlying ATP- And Adenosine Induced Microvascular Endothelial Barrier Preservationen_US
dc.typeOtheren
dc.contributor.departmentVascular Biology Centeren
dc.description.advisorVerin, Alexanderen
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