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dc.contributor.authorThunemann, Martinen_US
dc.contributor.authorSchmidt, Kjestineen_US
dc.contributor.authorde Wit, Coren_US
dc.contributor.authorHan, Xiaoxingen_US
dc.contributor.authorJain, Rakesh K.en_US
dc.contributor.authorFukumura, Daien_US
dc.contributor.authorFeil, Roberten_US
dc.coverage.spatialSwitzerlanden_US
dc.date2014-09-23
dc.date.accessioned2020-12-18T16:38:48Z
dc.date.available2020-12-18T16:38:48Z
dc.date.issued2014
dc.identifierhttps://www.ncbi.nlm.nih.gov/pubmed/25352809
dc.identifier.citationMartin Thunemann, Kjestine Schmidt, Cor de Wit, Xiaoxing Han, Rakesh K Jain, Dai Fukumura, Robert Feil. 2014. "Correlative intravital imaging of cGMP signals and vasodilation in mice.." Front Physiol, Volume 5, pp. 394 - ?. https://doi.org/10.3389/fphys.2014.00394
dc.identifier.issn1664-042X
dc.identifier.urihttps://hdl.handle.net/2144/41826
dc.description.abstractCyclic guanosine monophosphate (cGMP) is an important signaling molecule and drug target in the cardiovascular system. It is well known that stimulation of the vascular nitric oxide (NO)-cGMP pathway results in vasodilation. However, the spatiotemporal dynamics of cGMP signals themselves and the cGMP concentrations within specific cardiovascular cell types in health, disease, and during pharmacotherapy with cGMP-elevating drugs are largely unknown. To facilitate the analysis of cGMP signaling in vivo, we have generated transgenic mice that express fluorescence resonance energy transfer (FRET)-based cGMP sensor proteins. Here, we describe two models of intravital FRET/cGMP imaging in the vasculature of cGMP sensor mice: (1) epifluorescence-based ratio imaging in resistance-type vessels of the cremaster muscle and (2) ratio imaging by multiphoton microscopy within the walls of subcutaneous blood vessels accessed through a dorsal skinfold chamber. Both methods allow simultaneous monitoring of NO-induced cGMP transients and vasodilation in living mice. Detailed protocols of all steps necessary to perform and evaluate intravital imaging experiments of the vasculature of anesthetized mice including surgery, imaging, and data evaluation are provided. An image segmentation approach is described to estimate FRET/cGMP changes within moving structures such as the vessel wall during vasodilation. The methods presented herein should be useful to visualize cGMP or other biochemical signals that are detectable with FRET-based biosensors, such as cyclic adenosine monophosphate or Ca(2+), and to correlate them with respective vascular responses. With further refinement and combination of transgenic mouse models and intravital imaging technologies, we envision an exciting future, in which we are able to "watch" biochemistry, (patho-)physiology, and pharmacotherapy in the context of a living mammalian organism.en_US
dc.description.sponsorshipP01 CA080124 - NCI NIH HHS; R01 CA096915 - NCI NIH HHSen_US
dc.format.extentp. 394en_US
dc.languageeng
dc.language.isoen_US
dc.relation.ispartofFront Physiol
dc.rightsCopyright © 2014 Thunemann, Schmidt, de Wit, Han, Jain, Fukumura and Feil. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.en_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectBiosensoren_US
dc.subjectCremasteren_US
dc.subjectCyclic GMPen_US
dc.subjectDorsal skinfold chamberen_US
dc.subjectFluorescence resonance energy transferen_US
dc.subjectIntravital imagingen_US
dc.subjectMicrocirculationen_US
dc.subjectMultiphoton microscopyen_US
dc.subjectPhysiologyen_US
dc.subjectMedical physiologyen_US
dc.subjectPsychologyen_US
dc.titleCorrelative intravital imaging of cGMP signals and vasodilation in miceen_US
dc.typeArticleen_US
dc.description.versionPublished versionen_US
dc.identifier.doi10.3389/fphys.2014.00394
pubs.elements-sourcepubmeden_US
pubs.notesEmbargo: Not knownen_US
pubs.organisational-groupBoston Universityen_US
pubs.organisational-groupBoston University, College of Engineeringen_US
pubs.publication-statusPublished onlineen_US
dc.identifier.orcid0000-0003-4139-079X (Thunemann, Martin)
dc.identifier.mycv575738


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Copyright © 2014 Thunemann, Schmidt, de Wit, Han, Jain, Fukumura and Feil. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Except where otherwise noted, this item's license is described as Copyright © 2014 Thunemann, Schmidt, de Wit, Han, Jain, Fukumura and Feil. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.