Supplementary Material for: An Intact Kidney Slice Model to Investigate Vasa Recta Properties and Function in situ
2017
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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