Many questions still remain concerning both proximal tubule (PT) renin angiotensin system (RAS) intracellular configuration and mechanisms of its effects. This application uses two lines of immortalized rat PT cells (IRPTC) developed in this application's first cycle to define PT RAS paracrine, autocrine and intracrine effects. It is hypothesized that Ang II and other angiotensins produced locally within PT influence salt and water reabsorption by modulating transporters via specific intracellular signaling mechanisms and intranuclear mechanisms as yet incompletely deduced. Ang II appears to reduce cAMP and stimulate Ca2+ exchanger activity; to increase intracellular calcium via PKC and stimulate Na+/Ca++ transport; and to increase phospholipase A2, stimulating hydrolysis of phosphatidylcholine to lysophosphatidylcholine and fatty acid, influencing sodium transport. Such Ang II effects may be carried out both via surface and intracellular receptors in PT, as suggested by recent evidence from the applicant's laboratory that IRPTC have nuclear Ang II receptors.
Specific Aims are to define: I: Intracellular configuration and spatial arrangement of RAS in IRPTC. While all RAS components are clearly demonstrated in PT (and in both lines of IRPTC), actual intracellular localization is incompletely known. Intracellular production, localization and trafficking of RAS components will be defined by immunoelectron microscopy, confocal microscopy, subcellular fractionation plus molecular and cellular techniques. II: Mechanisms of Ang II effect on solute transport in PT. Ang II exhibits a biphasic influence on water and salt transport. The influence of Ang II on transporter regulation will be defined, using Na+/H+ exchanger as a prototype in parallel studies encompassing both molecular biology and cell physiology. Ang II-responsive elements will be sought; effects of Ang II on mRNA and protein regulation of NHEs will be delineated [e.g., mRNA steady state level, stability and localization; immunostaining intensity and pattern [e.g., with trafficking and immunoassay]; and cell signaling and physiology will be defined. III: Ang II and its autocrine and intracrine effects, as these affect PT functions such as growth and repair, and protein processing, hypothesizing that Ang II modulates cell growth and repair. The functional role of Ang II in IRPTCs will be defined by blocking or stimulating RAS components to dissect autocrine effects. IRPTCs have intracellular Ang II receptors; thus, possible intracellular processing of angiotensinogen (renin substrate) will be studied.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL048455-07
Application #
6183726
Study Section
Special Emphasis Panel (ZRG4-CVB (03))
Project Start
1992-05-01
Project End
2002-03-31
Budget Start
2000-04-01
Budget End
2002-03-31
Support Year
7
Fiscal Year
2000
Total Cost
$295,222
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
Zhao, Shuiling; Ghosh, Anindya; Lo, Chao-Sheng et al. (2018) Nrf2 Deficiency Upregulates Intrarenal Angiotensin-Converting Enzyme-2 and Angiotensin 1-7 Receptor Expression and Attenuates Hypertension and Nephropathy in Diabetic Mice. Endocrinology 159:836-852
Lo, Chao-Sheng; Shi, Yixuan; Chang, Shiao-Ying et al. (2015) Overexpression of heterogeneous nuclear ribonucleoprotein F stimulates renal Ace-2 gene expression and prevents TGF-?1-induced kidney injury in a mouse model of diabetes. Diabetologia 58:2443-54
Shi, Yixuan; Lo, Chao-Sheng; Padda, Ranjit et al. (2015) Angiotensin-(1-7) prevents systemic hypertension, attenuates oxidative stress and tubulointerstitial fibrosis, and normalizes renal angiotensin-converting enzyme 2 and Mas receptor expression in diabetic mice. Clin Sci (Lond) 128:649-63
Abdo, Shaaban; Shi, Yixuan; Otoukesh, Abouzar et al. (2014) Catalase overexpression prevents nuclear factor erythroid 2-related factor 2 stimulation of renal angiotensinogen gene expression, hypertension, and kidney injury in diabetic mice. Diabetes 63:3483-96
Shi, Yixuan; Lo, Chao-Sheng; Chenier, Isabelle et al. (2013) Overexpression of catalase prevents hypertension and tubulointerstitial fibrosis and normalization of renal angiotensin-converting enzyme-2 expression in Akita mice. Am J Physiol Renal Physiol 304:F1335-46
Abdo, S; Lo, C-S; Chenier, I et al. (2013) Heterogeneous nuclear ribonucleoproteins F and K mediate insulin inhibition of renal angiotensinogen gene expression and prevention of hypertension and kidney injury in diabetic mice. Diabetologia 56:1649-60
Lo, Chao-Sheng; Chang, Shiao-Ying; Chenier, Isabelle et al. (2012) Heterogeneous nuclear ribonucleoprotein F suppresses angiotensinogen gene expression and attenuates hypertension and kidney injury in diabetic mice. Diabetes 61:2597-608
Lau, Garnet J; Godin, Nicolas; Maachi, Hasna et al. (2012) Bcl-2-modifying factor induces renal proximal tubular cell apoptosis in diabetic mice. Diabetes 61:474-84
Lo, Chao-Sheng; Liu, Fang; Shi, Yixuan et al. (2012) Dual RAS blockade normalizes angiotensin-converting enzyme-2 expression and prevents hypertension and tubular apoptosis in Akita angiotensinogen-transgenic mice. Am J Physiol Renal Physiol 302:F840-52
Godin, Nicolas; Liu, Fang; Lau, Garnet J et al. (2010) Catalase overexpression prevents hypertension and tubular apoptosis in angiotensinogen transgenic mice. Kidney Int 77:1086-97

Showing the most recent 10 out of 57 publications