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An Intact Kidney Slice Model to Investigate Vasa Recta Properties and Function in situ

BACKGROUND: Medullary blood flow is via vasa recta capillaries, which possess contractile pericytes. In vitro studies using isolated descending vasa recta show that pericytes can constrict/dilate descending vasa recta when vasoactive substances are present. We describe a live kidney slice model in w...

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Autores principales: Crawford, C., Kennedy-Lydon, T., Sprott, C., Desai, T., Sawbridge, L., Munday, J., Unwin, R.J., Wildman, S.S.P., Peppiatt-Wildman, C.M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: S. Karger AG 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5166522/
https://www.ncbi.nlm.nih.gov/pubmed/22833057
http://dx.doi.org/10.1159/000339110
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author Crawford, C.
Kennedy-Lydon, T.
Sprott, C.
Desai, T.
Sawbridge, L.
Munday, J.
Unwin, R.J.
Wildman, S.S.P.
Peppiatt-Wildman, C.M.
author_facet Crawford, C.
Kennedy-Lydon, T.
Sprott, C.
Desai, T.
Sawbridge, L.
Munday, J.
Unwin, R.J.
Wildman, S.S.P.
Peppiatt-Wildman, C.M.
author_sort Crawford, C.
collection PubMed
description BACKGROUND: Medullary blood flow is via vasa recta capillaries, which possess contractile pericytes. In vitro studies using isolated descending vasa recta show that pericytes can constrict/dilate descending vasa recta when vasoactive substances are present. We describe a live kidney slice model in which pericyte-mediated vasa recta constriction/dilation can be visualized in situ. METHODS: Confocal microscopy was used to image calcein, propidium iodide and Hoechst labelling in ‘live’ kidney slices, to determine tubular and vascular cell viability and morphology. DIC video-imaging of live kidney slices was employed to investigate pericyte-mediated real-time changes in vasa recta diameter. RESULTS: Pericytes were identified on vasa recta and their morphology and density were characterized in the medulla. Pericyte-mediated changes in vasa recta diameter (10–30%) were evoked in response to bath application of vasoactive agents (norepinephrine, endothelin-1, angiotensin-II and prostaglandin E(2)) or by manipulating endogenous vasoactive signalling pathways (using tyramine, L-NAME, a cyclo-oxygenase (COX-1) inhibitor indomethacin, and ATP release). CONCLUSIONS: The live kidney slice model is a valid complementary technique for investigating vasa recta function in situ and the role of pericytes as regulators of vasa recta diameter. This technique may also be useful in exploring the role of tubulovascular crosstalk in regulation of medullary blood flow.
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spelling pubmed-51665222017-03-08 An Intact Kidney Slice Model to Investigate Vasa Recta Properties and Function in situ Crawford, C. Kennedy-Lydon, T. Sprott, C. Desai, T. Sawbridge, L. Munday, J. Unwin, R.J. Wildman, S.S.P. Peppiatt-Wildman, C.M. Nephron Physiol Original Paper BACKGROUND: Medullary blood flow is via vasa recta capillaries, which possess contractile pericytes. In vitro studies using isolated descending vasa recta show that pericytes can constrict/dilate descending vasa recta when vasoactive substances are present. We describe a live kidney slice model in which pericyte-mediated vasa recta constriction/dilation can be visualized in situ. METHODS: Confocal microscopy was used to image calcein, propidium iodide and Hoechst labelling in ‘live’ kidney slices, to determine tubular and vascular cell viability and morphology. DIC video-imaging of live kidney slices was employed to investigate pericyte-mediated real-time changes in vasa recta diameter. RESULTS: Pericytes were identified on vasa recta and their morphology and density were characterized in the medulla. Pericyte-mediated changes in vasa recta diameter (10–30%) were evoked in response to bath application of vasoactive agents (norepinephrine, endothelin-1, angiotensin-II and prostaglandin E(2)) or by manipulating endogenous vasoactive signalling pathways (using tyramine, L-NAME, a cyclo-oxygenase (COX-1) inhibitor indomethacin, and ATP release). CONCLUSIONS: The live kidney slice model is a valid complementary technique for investigating vasa recta function in situ and the role of pericytes as regulators of vasa recta diameter. This technique may also be useful in exploring the role of tubulovascular crosstalk in regulation of medullary blood flow. S. Karger AG 2012-08 2012-07-20 /pmc/articles/PMC5166522/ /pubmed/22833057 http://dx.doi.org/10.1159/000339110 Text en Copyright © 2012 by S. Karger AG, Basel http://creativecommons.org/licenses/by/4.0/ This article is licensed under the Creative Commons Attribution 4.0 International License (CC BY) (http://www.karger.com/Services/OpenAccessLicense). Usage, derivative works and distribution are permitted provided that proper credit is given to the author and the original publisher.
spellingShingle Original Paper
Crawford, C.
Kennedy-Lydon, T.
Sprott, C.
Desai, T.
Sawbridge, L.
Munday, J.
Unwin, R.J.
Wildman, S.S.P.
Peppiatt-Wildman, C.M.
An Intact Kidney Slice Model to Investigate Vasa Recta Properties and Function in situ
title An Intact Kidney Slice Model to Investigate Vasa Recta Properties and Function in situ
title_full An Intact Kidney Slice Model to Investigate Vasa Recta Properties and Function in situ
title_fullStr An Intact Kidney Slice Model to Investigate Vasa Recta Properties and Function in situ
title_full_unstemmed An Intact Kidney Slice Model to Investigate Vasa Recta Properties and Function in situ
title_short An Intact Kidney Slice Model to Investigate Vasa Recta Properties and Function in situ
title_sort intact kidney slice model to investigate vasa recta properties and function in situ
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5166522/
https://www.ncbi.nlm.nih.gov/pubmed/22833057
http://dx.doi.org/10.1159/000339110
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