Our working hypothesis is that the local control of tissue blood flow reflects the coordination of activity among endothelial cells and smooth nnuscle cells of microvascular resistance networks.
The Specific Aims of this project focus on understanding how "rapid" (electrical) and "slow" (calcium waves) components of conducted vasodilation are initiated, how they travel from cell to cell, and how they interact with each other. In accord with the long term nature and innovative scope of this MERIT Award, we have invested substantively in developing novel methods to study these relationships. In freshly isolated endothelial tubes of feed arteries from mouse skeletal muscle, dual simultaneous intracellular recording demonstrates that electrical conduction along the endothelium through gap junctions can be tuned by governing charge loss through potassium channels in the plasma membrane. Complementary experiments show that glycyrrhetinic acid derivatives used widely by others to inhibit gap junctions concomitantly block potassium channels, disqualifying these agents in resolving respective determinants of electrical signaling. We quantified calcium signaling in endothelial tubes using Fura-2 photometry, establishing a foundation for implementing confocal imaging and multi-photon technology which we are using to investigate mechansims of calcium signaling within and between individual cells along the endothelium. In developing intravital macrozoom imaging of transgenic mice that express the calcium-sensitive protein GCaMP2 in arteriolar endothelium, we determined that the initiation of calcium waves with acetylcholine microiontophoresis was independent of myogenic tone and distinguished between the local initation of conducted responses from more global effects when the agonist gained access to the flow stream. In light of studies of myoendothelial coupling that have been based entirely upon isolated preparations studied in vitro, we are investigating endothelial calcium signaling in response to neural activation of smooth muscle cells in vivo. We plan to continue pursuing the Specific Aims of this project in light of our new methodology and findings. Our long-term goals center on defining the signaling events which coordinate the activity of endothelium and smooth muscle in resistance vessels that control the delivery of oxygen and nutrients to tissue cells. Resolving these relationships will provide critical new insight for determining how respective signaling pathways may be affected (and thereby treated) during conditions known to alter endothelial and vascular smooth muscle function including widespread diseased states of obesity, diabetes and hypertension.

Public Health Relevance

The goal of this research project is to understand how electrical and calcium signals coordinate cells of the blood vessel wall to increase ttssue blood flow and oxygen delivery. Understanding how vasodilator signals originate and are coordinated in resistance networks provides new insight for developing novel therapies for treating diseases associated with vascular complications and impaired tissue perfusion.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37HL041026-21
Application #
8479592
Study Section
Special Emphasis Panel (NSS)
Program Officer
OH, Youngsuk
Project Start
1988-07-01
Project End
2019-02-28
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
21
Fiscal Year
2014
Total Cost
$362,416
Indirect Cost
$117,327
Name
University of Missouri-Columbia
Department
Pharmacology
Type
Schools of Medicine
DUNS #
153890272
City
Columbia
State
MO
Country
United States
Zip Code
65211
Sinkler, Shenghua Y; Fernando, Charmain A; Segal, Steven S (2016) Differential α-adrenergic modulation of rapid onset vasodilatation along resistance networks of skeletal muscle in old versus young mice. J Physiol 594:6987-7004
Boerman, Erika M; Segal, Steven S (2016) Depressed perivascular sensory innervation of mouse mesenteric arteries with advanced age. J Physiol 594:2323-38
Boerman, Erika M; Everhart, Jesse E; Segal, Steven S (2016) Advanced age decreases local calcium signaling in endothelium of mouse mesenteric arteries in vivo. Am J Physiol Heart Circ Physiol 310:H1091-6
Fernando, Charmain A; Liu, Yajun; Sowa, Grzegorz et al. (2016) Attenuated rapid onset vasodilation with greater force production in skeletal muscle of caveolin-2-/- mice. Am J Physiol Heart Circ Physiol 311:H415-25
Segal, Steven S (2015) Integration and Modulation of Intercellular Signaling Underlying Blood Flow Control. J Vasc Res 52:136-57
Nance, Michael E; Whitfield, Justin T; Zhu, Yi et al. (2015) Attenuated sarcomere lengthening of the aged murine left ventricle observed using two-photon fluorescence microscopy. Am J Physiol Heart Circ Physiol 309:H918-25
Behringer, Erik J; Segal, Steven S (2015) Membrane potential governs calcium influx into microvascular endothelium: integral role for muscarinic receptor activation. J Physiol 593:4531-48
Socha, Matthew J; Boerman, Erika M; Behringer, Erik J et al. (2015) Advanced age protects microvascular endothelium from aberrant Ca(2+) influx and cell death induced by hydrogen peroxide. J Physiol 593:2155-69
Segal, Steven S (2014) Blood flow restriction without sympathetic vasoconstriction in ageing skeletal muscle during exercise. J Physiol 592:4607-8
Domeier, Timothy L; Roberts, Cale J; Gibson, Anne K et al. (2014) Dantrolene suppresses spontaneous Ca2+ release without altering excitation-contraction coupling in cardiomyocytes of aged mice. Am J Physiol Heart Circ Physiol 307:H818-29

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