The long-term goal of this study is to elucidate the regulation of renal hemodynamics by the endothelial (ECs) and smooth muscle cells (SMCs) acting as a functional unit. The focus of the proposed study is on the possible feed-forward and feedback interactions between renal microvascular ECs and SMCs with regard to the production and action of nitric oxide (NO) and endothelin (ET). The major hypothesis to be tested is based on original observations suggesting the possibility of the oscillatory drive intrinsic to the integrated ECs-SMCs unit.
The specific aims of the proposed study comprise investigations of the operation mode of the ECs-SMCs unit by exploring the system at various levels of organization, from molecular genetics to whole organ function, as follows: 1. To elucidate the functional role of the ET/B receptor in ECs. 2. To elucidate the role of intracellular messengers and physical forces acting on ECs in regulating the activity of constitutive NO synthase. 3. To determine whether physiological contraction of SMCs activates ECs production of NO and ET-1, and the possible mediators of this action. 4. To elucidate the contribution of deformation-related factors (stretch, pressure) and flow-related factors (shear) on the time-course of continuously and simultaneously monitored ET-1 and NO synthesis. Toward accomplishing the goals of the study, several novel technical approaches were developed: 1) The application of an NO-selective microelectrode to continuously monitor NO release from ECs and perfused resistance vessels; 2) The synthesis of the site-specific biotinylated ET- 1 and biotinylated IRL-1620 which allow to perform vital fluorescence mapping of ET/A and ET/B receptors; 3) The establishment of immortalized ECs and SMCs from rat renal resistance arteries; 4) The establishment of genetically-engineered Chinese hamster ovary cells stably expressing the ET/B receptor and/or endothelial NO synthase; 5) The development of a """"""""sandwich"""""""" coculture system to model the interaction between the ECs and SMCs. Other approaches to be used in the proposed study include videomicroscopy of renal microvasculature, image analysis of cell shape changes and cytosolic calcium concentration, quantitative fluorescence mapping of ET receptors, and radioimmunoassay and bioassay for ET-1. These studies will elucidate the role of physical factors and sensory inputs regulating ECs production of NO and ET-1, according to the proposed """"""""ping-pong"""""""" model, feedback regulation of ECs function by contracting SMCs, and the capacity of the integrated ECs-SMCs unit for auto- oscillations.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK045462-03
Application #
2518326
Study Section
General Medicine B Study Section (GMB)
Project Start
1995-09-01
Project End
1999-08-31
Budget Start
1997-09-01
Budget End
1998-08-31
Support Year
3
Fiscal Year
1997
Total Cost
Indirect Cost
Name
State University New York Stony Brook
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
804878247
City
Stony Brook
State
NY
Country
United States
Zip Code
11794
Matsumoto, Kei; Xavier, Sandhya; Chen, Jun et al. (2017) Instructive Role of the Microenvironment in Preventing Renal Fibrosis. Stem Cells Transl Med 6:992-1005
Song, J W; Zullo, J A; Liveris, D et al. (2017) Therapeutic Restoration of Endothelial Glycocalyx in Sepsis. J Pharmacol Exp Ther 361:115-121
Lipphardt, Mark; Song, Jong W; Matsumoto, Kei et al. (2017) The third path of tubulointerstitial fibrosis: aberrant endothelial secretome. Kidney Int 92:558-568
Goligorsky, M S; Hirschi, K (2016) Stress-Induced Premature Senescence of Endothelial and Endothelial Progenitor Cells. Adv Pharmacol 77:281-306
Kida, Yujiro; Zullo, Joseph A; Goligorsky, Michael S (2016) Endothelial sirtuin 1 inactivation enhances capillary rarefaction and fibrosis following kidney injury through Notch activation. Biochem Biophys Res Commun 478:1074-9
Zullo, Joseph A; Fan, Jie; Azar, Tala T et al. (2016) Exocytosis of Endothelial Lysosome-Related Organelles Hair-Triggers a Patchy Loss of Glycocalyx at the Onset of Sepsis. Am J Pathol 186:248-58
Lin, Chi Hua Sarah; Chen, Jun; Zhang, Zhongtao et al. (2016) Endostatin and transglutaminase 2 are involved inĀ fibrosis of the aging kidney. Kidney Int 89:1281-92
Kida, Yujiro; Goligorsky, Michael S (2016) Sirtuins, Cell Senescence, and Vascular Aging. Can J Cardiol 32:634-41
Zullo, Joseph A; Nadel, Ellen P; Rabadi, May M et al. (2015) The Secretome of Hydrogel-Coembedded Endothelial Progenitor Cells and Mesenchymal Stem Cells Instructs Macrophage Polarization in Endotoxemia. Stem Cells Transl Med 4:852-61
Zullo, Joseph; Matsumoto, Kei; Xavier, Sandhya et al. (2015) The cell secretome, a mediator of cell-to-cell communication. Prostaglandins Other Lipid Mediat 120:17-20

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