Journal article
Intracellular beta(1)-Adrenergic Receptors and Organic Cation Transporter 3 Mediate Phospholamban Phosphorylation to Enhance Cardiac Contractility
Circulation research, Vol.128(2), pp.246-261
01/22/2021
DOI: 10.1161/CIRCRESAHA.120.317452
PMCID: PMC7856104
PMID: 33183171
Abstract
Rationale:
beta(1)ARs (beta(1)-adrenoceptors) exist at intracellular membranes and OCT3 (organic cation transporter 3) mediates norepinephrine entry into cardiomyocytes. However, the functional role of intracellular beta(1)AR in cardiac contractility remains to be elucidated.
Objective:
Test localization and function of intracellular beta(1)AR on cardiac contractility.
Methods and Results:
Membrane fractionation, super-resolution imaging, proximity ligation, coimmunoprecipitation, and single-molecule pull-down demonstrated a pool of beta(1)ARs in mouse hearts that were associated with sarco/endoplasmic reticulum Ca2+-ATPase at the sarcoplasmic reticulum (SR). Local PKA (protein kinase A) activation was measured using a PKA biosensor targeted at either the plasma membrane (PM) or SR. Compared with wild-type, myocytes lacking OCT3 (OCT3-KO [OCT3 knockout]) responded identically to the membrane-permeant beta AR agonist isoproterenol in PKA activation at both PM and SR. The same was true at the PM for membrane-impermeant norepinephrine, but the SR response to norepinephrine was suppressed in OCT3-KO myocytes. This differential effect was recapitulated in phosphorylation of the SR-pump regulator phospholamban. Similarly, OCT3-KO selectively suppressed calcium transients and contraction responses to norepinephrine but not isoproterenol. Furthermore, sotalol, a membrane-impermeant beta AR-blocker, suppressed isoproterenol-induced PKA activation at the PM but permitted PKA activation at the SR, phospholamban phosphorylation, and contractility. Moreover, pretreatment with sotalol in OCT3-KO myocytes prevented norepinephrine-induced PKA activation at both PM and the SR and contractility.
Conclusions:
Functional beta(1)ARs exists at the SR and is critical for PKA-mediated phosphorylation of phospholamban and cardiac contractility upon catecholamine stimulation. Activation of these intracellular beta(1)ARs requires catecholamine transport via OCT3.
Details
- Title: Subtitle
- Intracellular beta(1)-Adrenergic Receptors and Organic Cation Transporter 3 Mediate Phospholamban Phosphorylation to Enhance Cardiac Contractility
- Creators
- Ying Wang - University of California, DavisQian Shi - University of California, DavisMinghui Li - University of California, DavisMeimi Zhao - University of California, DavisRaghavender Reddy Gopireddy - University of California, DavisJian-Peng Teoh - University of California, DavisBing Xu - University of California, DavisChaoqun Zhu - University of California, DavisKyle E. Ireton - University of California, DavisSanghavi Srinivasan - University of California, DavisShaoliang Chen - Nanjing Medical UniversityPaul J. Gasser - Marquette UniversityJulie Bossuyt - University of California, DavisJohannes W. Hell - University of California, DavisDonald M. Bers - University of California, DavisYang K. Xiang - University of California, Davis
- Resource Type
- Journal article
- Publication Details
- Circulation research, Vol.128(2), pp.246-261
- Publisher
- Lippincott Williams & Wilkins
- DOI
- 10.1161/CIRCRESAHA.120.317452
- PMID
- 33183171
- PMCID
- PMC7856104
- ISSN
- 0009-7330
- eISSN
- 1524-4571
- Number of pages
- 16
- Grant note
- American Heart Association postdoctoral fellowship; American Heart Association R01-HL127764; R01-HL147263; R01-HL133832; P01-HL141084; R01 NS078792; R01-AG055357; R01-MH097887 / National Institutes of Health; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA 01BX002900 / VA Merit grant 81700252 / National Natural Science Foundation of China; National Natural Science Foundation of China (NSFC)
- Language
- English
- Date published
- 01/22/2021
- Academic Unit
- Endocrinology and Metabolism; Internal Medicine
- Record Identifier
- 9984359808702771
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