Increased microvascular permeability is a hallmark of inflammation. Previous research has focused on how to prevent the onset and the maintenance of the elevated permeability under inflammatory conditions. . We propose to test the central hypothesis that Microvascular permeability is regulated by counterbalancing signaling mechanisms that maintain homeostasis by enhancing permeability or barrier properties, respectively. We will investigate eNOS translocation as a mechanism that stimulates hyperpermeability and mechanisms that inactivate hyperpermeability in the early inflammatory phase. We will study these processes in endothelial cells and in striated muscle. We hypothesize that eNOS translocation is necessary to achieve effective NO concentrations at or near the effectors for permeability. We also hypothesize that at or near the peak of the hyperpermeability response, the endothelium (or tissue) begins a process to inactivate or reverse hyperpermeability and restore the physiological barrier properties of the microvascular wall. We further propose that the small GTP-binding proteins Epac/Rap-1 serve as `Barrier Enhancing Factors'and participate in the hyperpermeability-inactivation/reversal process. The Specific Hypotheses and Specific Aims to be tested are: SPECIFIC HYPOTHESIS 1. eNOS translocation to cytosol (non-Golgi cytosol compartment) is a step in endothelial regulation of microvascular permeability.
SPECIFIC AIM 1. To investigate the relationship between eNOS translocation and regulation of endothelial and microvascular permeability. SPECIFIC HYPOTHESIS 2: cAMP via Epac inactivates or reverses agonist-induced hyperpermeability.
SPECIFIC AIM 2. 1 To investigate the timed inactivation/reversal of hyperpermeability.
SPECIFIC AIM 2. 1. To determine whether or not stimulation of Epac reverses (inactivates) agonist-induced hyperpermeability. We will apply intravital microscopy in conjunction with computer-assisted image analysis and molecular biology approaches to elucidate the postulated mechanisms.

Public Health Relevance

A better understanding of the molecular mechanisms and of the timing between hyperpermeability and its """"""""physiological"""""""" inactivation or reversal in inflammatory processes should provide a window of opportunity for promoting inactivation or reversal of hyperpermeability to prevent complications such as excessive edema and compartment syndrome in (micro)vascular disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL070634-08
Application #
7997190
Study Section
Hypertension and Microcirculation Study Section (HM)
Program Officer
Thrasher, Terry N
Project Start
2002-07-01
Project End
2012-12-31
Budget Start
2011-01-01
Budget End
2011-12-31
Support Year
8
Fiscal Year
2011
Total Cost
$408,600
Indirect Cost
Name
University of Medicine & Dentistry of NJ
Department
Pharmacology
Type
Schools of Medicine
DUNS #
623946217
City
Newark
State
NJ
Country
United States
Zip Code
07107
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Duran, Walter N; Breslin, Jerome W; Sanchez, Fabiola A (2010) The NO cascade, eNOS location, and microvascular permeability. Cardiovasc Res 87:254-61
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Sanchez, Fabiola A; Rana, Roshniben; Kim, David D et al. (2009) Internalization of eNOS and NO delivery to subcellular targets determine agonist-induced hyperpermeability. Proc Natl Acad Sci U S A 106:6849-53
Kim, David D; Kanetaka, Takehito; Duran, Ricardo G et al. (2009) Independent regulation of periarteriolar and perivenular nitric oxide mechanisms in the in vivo hamster cheek pouch microvasculature. Microcirculation 16:323-30

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