Salt retention and consequent plasma volume expansion inevitably cause elevation of blood pressure (BP) which, in the chronic state (hypertension), is sustained by increased total peripheral vascular resistance (TPR). Surprisingly, the "signaling pathway" by which salt retention increases TPR and BP is unresolved. Our goal is to elucidate the specific molecular mechanisms that link salt retention to hypertension using mouse models because salt-dependent hypertension is similar in humans and rodents. We will employ pharmacological tools and genetically-engineered mice to test the hypothesis that: i) Endogenous ouabain (EO), ii) Inhibited activity of arterial Na+ pumps with a (ouabain-sensitive, S) ?2 catalytic subunit, and iii) Increased Ca2+ entry via arterial Na-Ca exchanger-1 (NCX1) are key steps in the signaling pathway.
The Specific Aims address three sub-hypotheses: 1) That salt- dependent hypertension in mice with transgenic G-protein coupled receptor kinase-4 [GRK4?(A486V)] is mediated by EO, ?2 Na+ pumps, and NCX1. The effects of dietary salt on plasma EO and BP will be measured in non-transgenic GRK4? and salt-sensitive GRK4?(A486V) mice. The effects of Digibind (antibodies that bind ouabain), PST-2238 (ouabain antagonist) and SEA0400 (NCX1 blocker), and of knock-down of ?2 Na+ pump or reduced NCX1 expression, on the salt-dependent rise in BP (?BP) will be determined. 2) That reduced ?2 Na+ pump expression increases salt-sensitivity, and that salt- dependent ?BP is mediated by EO acting on ?2 Na+ pumps, and by NCX1. The effects of dietary salt on plasma EO and BP will be measured in WT (?2S/S) mice and mice with one null mutant ?2 allele (?2S/-). The effects of Digibind, PST-2238, SEA0400 and reduced NCX1 expression will be tested on the salt- dependent ?BP. 3) That salt sensitivity due to smooth muscle (sm)-specific NCX1.3 overexpression correlates with hypersensitivity to ouabain and is augmented by reduced ?2 Na+ pump abundance. The effects of dietary salt on plasma EO and BP will be measured in WT mice and mice that overexpress smNCX1.3 (NCX1.3smTg/Tg);the effects of Digibind and PST-2238, and of knock-down of sm-?2 pump expression, will be tested on salt-dependent ?BP. The effects of ouabain and angiotensin II infusion on BP will be compared in WT, NCX1.3smTg/Tg, and sm-specific NCX1 knockout mice. The results will elucidate some key mechanisms that link salt to hypertension, will provide a roadmap for human studies, and will pinpoint new therapeutic targets.

Public Health Relevance

The goal of this project is to determine the specific mechanisms by which ingestion of excess salt raises blood pressure. We will study proposed steps in the pathway that links salt to hypertension by using genetically-modified mice in which these steps are augmented or suppressed. The results will elucidate key mechanisms that participate in this salt-hypertension pathway in humans as well as in rodents, will provide a roadmap for human studies, and will identify new targets for drug therapy.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL045215-25
Application #
8267671
Study Section
Hypertension and Microcirculation Study Section (HM)
Program Officer
OH, Youngsuk
Project Start
1990-07-01
Project End
2014-05-31
Budget Start
2012-06-01
Budget End
2014-05-31
Support Year
25
Fiscal Year
2012
Total Cost
$407,924
Indirect Cost
$111,363
Name
University of Maryland Baltimore
Department
Physiology
Type
Schools of Medicine
DUNS #
188435911
City
Baltimore
State
MD
Country
United States
Zip Code
21201
Blaustein, Mordecai P (2014) Why isn't endogenous ouabain more widely accepted? Am J Physiol Heart Circ Physiol 307:H635-9
Hamlyn, John M; Linde, Cristina I; Gao, Junjie et al. (2014) Neuroendocrine humoral and vascular components in the pressor pathway for brain angiotensin II: a new axis in long term blood pressure control. PLoS One 9:e108916
Song, Hong; Karashima, Eiji; Hamlyn, John M et al. (2014) Ouabain-digoxin antagonism in rat arteries and neurones. J Physiol 592:941-69
Song, Hong; Thompson, Scott M; Blaustein, Mordecai P (2013) Nanomolar ouabain augments Ca2+ signalling in rat hippocampal neurones and glia. J Physiol 591:1671-89
Hamlyn, John M; Blaustein, Mordecai P (2013) Salt sensitivity, endogenous ouabain and hypertension. Curr Opin Nephrol Hypertens 22:51-8
Blaustein, Mordecai P (2013) Livin' with NCX and lovin' it: a 45 year romance. Adv Exp Med Biol 961:3-15
Zulian, Alessandra; Linde, Cristina I; Pulina, Maria V et al. (2013) Activation of c-SRC underlies the differential effects of ouabain and digoxin on Ca(2+) signaling in arterial smooth muscle cells. Am J Physiol Cell Physiol 304:C324-33
Pulina, Maria V; Zulian, A; Baryshnikov, Sergey G et al. (2013) Cross talk between plasma membrane Na(+)/Ca (2+) exchanger-1 and TRPC/Orai-containing channels: key players in arterial hypertension. Adv Exp Med Biol 961:365-74
Blaustein, Mordecai P; Leenen, Frans H H; Chen, Ling et al. (2012) How NaCl raises blood pressure: a new paradigm for the pathogenesis of salt-dependent hypertension. Am J Physiol Heart Circ Physiol 302:H1031-49
Jacobs, Brandiese E; Liu, Yong; Pulina, Maria V et al. (2012) Normal pregnancy: mechanisms underlying the paradox of a ouabain-resistant state with elevated endogenous ouabain, suppressed arterial sodium calcium exchange, and low blood pressure. Am J Physiol Heart Circ Physiol 302:H1317-29

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