Functional LTCC-β2AR Complex Needs Caveolin-3 and Is Disrupted in Heart Failure

Jose l. Sanchez-Alonso*; Laura Fedele*; Jaël s. Copier; Carla Lucarelli; Catherine Mansfield; Aleksandra Judina; Steven r. Houser; Thomas Brand; Julia Gorelik

doi:10.1161/CIRCRESAHA.123.322508

Functional LTCC-β2AR Complex Needs Caveolin-3 and Is Disrupted in Heart Failure

Jose l. Sanchez-Alonso*, Laura Fedele*, Jaël s. Copier, Carla Lucarelli, Catherine Mansfield, Aleksandra Judina, Steven r. Houser, Thomas Brand, Julia Gorelik

Wolfson SPaRC

Department of Physiology, Cardiovascular Research Center, Lewis Katz Temple University School of Medicine, Philadelphia, PA (S.R.H.).
National Heart and Lung Institute, Imperial College London, United Kingdom (J.L.S.-A., L.F., J.S.C., C.L., C.M., A.J., T.B., J.G.).

Research output: Contribution to journal › Article › peer-review

9 Citations (Scopus)

Abstract

Background: Beta-2 adrenergic receptors (β2ARs) but not beta-2 adrenergic receptors (β1ARs) form a functional complex with L-type Ca2+ channels (LTCCs) on the cardiomyocyte membrane. However, how microdomain localization in the plasma membrane affects the function of these complexes is unknown. We aim to study the coupling between LTCC and β adrenergic receptors in different cardiomyocyte microdomains, the distinct involvement of PKA and CAMKII (Ca2+/calmodulin-dependent protein kinase II) and explore how this functional complex is disrupted in heart failure. Methods: Global signaling between LTCCs and β adrenergic receptors was assessed with whole-cell current recordings and western blot analysis. Super-resolution scanning patch-clamp was used to explore the local coupling between single LTCCs and β1AR or β2AR in different membrane microdomains in control and failing cardiomyocytes. Results: LTCC open probability (Po) showed an increase from 0.054±0.003 to 0.092±0.008 when β2AR was locally stimulated in the proximity of the channel (<350 nm) in the transverse tubule microdomain. In failing cardiomyocytes, from both rodents and humans, this transverse tubule coupling between LTCC and β2AR was lost. Interestingly, local stimulation of β1AR did not elicit any change in the Po of LTCCs, indicating a lack of proximal functional interaction between the two, but we confirmed a general activation of LTCC via β1AR. By using blockers of PKA and CaMKII and a Caveolin-3-knockout mouse model, we conclude that the β2AR-LTCC regulation requires the presence of caveolin-3 and the activation of the CaMKII pathway. By contrast, at a cellular "global"level PKA plays a major role downstream β1AR and results in an increase in LTCC current. Conclusions: Regulation of the LTCC activity by proximity coupling mechanisms occurs only via β2AR, but not β1AR. This may explain how β2ARs tune the response of LTCCs to adrenergic stimulation in healthy conditions. This coupling is lost in heart failure; restoring it could improve the adrenergic response of failing cardiomyocytes.

Original language	English
Pages (from-to)	120-137
Number of pages	18
Journal	Circulation Research
Volume	133
Issue number	2
DOIs	https://doi.org/10.1161/CIRCRESAHA.123.322508
Publication status	Published - 7 Jul 2023

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10.1161/CIRCRESAHA.123.322508

https://www.ahajournals.org/doi/10.1161/CIRCRESAHA.123.322508

Cite this

@article{4648a22e3fee4b5d8eb36e228e47eb41,

title = "Functional LTCC-β2AR Complex Needs Caveolin-3 and Is Disrupted in Heart Failure",

abstract = "Background: Beta-2 adrenergic receptors (β2ARs) but not beta-2 adrenergic receptors (β1ARs) form a functional complex with L-type Ca2+ channels (LTCCs) on the cardiomyocyte membrane. However, how microdomain localization in the plasma membrane affects the function of these complexes is unknown. We aim to study the coupling between LTCC and β adrenergic receptors in different cardiomyocyte microdomains, the distinct involvement of PKA and CAMKII (Ca2+/calmodulin-dependent protein kinase II) and explore how this functional complex is disrupted in heart failure. Methods: Global signaling between LTCCs and β adrenergic receptors was assessed with whole-cell current recordings and western blot analysis. Super-resolution scanning patch-clamp was used to explore the local coupling between single LTCCs and β1AR or β2AR in different membrane microdomains in control and failing cardiomyocytes. Results: LTCC open probability (Po) showed an increase from 0.054±0.003 to 0.092±0.008 when β2AR was locally stimulated in the proximity of the channel (<350 nm) in the transverse tubule microdomain. In failing cardiomyocytes, from both rodents and humans, this transverse tubule coupling between LTCC and β2AR was lost. Interestingly, local stimulation of β1AR did not elicit any change in the Po of LTCCs, indicating a lack of proximal functional interaction between the two, but we confirmed a general activation of LTCC via β1AR. By using blockers of PKA and CaMKII and a Caveolin-3-knockout mouse model, we conclude that the β2AR-LTCC regulation requires the presence of caveolin-3 and the activation of the CaMKII pathway. By contrast, at a cellular {"}global{"}level PKA plays a major role downstream β1AR and results in an increase in LTCC current. Conclusions: Regulation of the LTCC activity by proximity coupling mechanisms occurs only via β2AR, but not β1AR. This may explain how β2ARs tune the response of LTCCs to adrenergic stimulation in healthy conditions. This coupling is lost in heart failure; restoring it could improve the adrenergic response of failing cardiomyocytes.",

author = "Sanchez-Alonso*, {Jose l.} and Laura Fedele* and Copier, {Ja{\"e}l s.} and Carla Lucarelli and Catherine Mansfield and Aleksandra Judina and Houser, {Steven r.} and Thomas Brand and Julia Gorelik",

note = "Funding Information: This work was supported by British Heart Foundation (grant RG/17/13/33173 to Dr Gorelik, grant PG/19/13/34247C to Dr Brand/Dr Gorelik, and grant RG/F/22/110081 to Dr Gorelik/Dr Sanchez-Alonso). Publisher Copyright: {\textcopyright} 2023 The Authors.",

year = "2023",

month = jul,

day = "7",

doi = "10.1161/CIRCRESAHA.123.322508",

language = "English",

volume = "133",

pages = "120--137",

journal = "Circulation Research",

issn = "0009-7330",

publisher = "Lippincott Williams and Wilkins",

number = "2",

}

TY - JOUR

T1 - Functional LTCC-β2AR Complex Needs Caveolin-3 and Is Disrupted in Heart Failure

AU - Sanchez-Alonso, Jose l.

AU - Fedele, Laura

AU - Copier, Jaël s.

AU - Lucarelli, Carla

AU - Mansfield, Catherine

AU - Judina, Aleksandra

AU - Houser, Steven r.

AU - Brand, Thomas

AU - Gorelik, Julia

N1 - Funding Information: This work was supported by British Heart Foundation (grant RG/17/13/33173 to Dr Gorelik, grant PG/19/13/34247C to Dr Brand/Dr Gorelik, and grant RG/F/22/110081 to Dr Gorelik/Dr Sanchez-Alonso). Publisher Copyright: © 2023 The Authors.

PY - 2023/7/7

Y1 - 2023/7/7

N2 - Background: Beta-2 adrenergic receptors (β2ARs) but not beta-2 adrenergic receptors (β1ARs) form a functional complex with L-type Ca2+ channels (LTCCs) on the cardiomyocyte membrane. However, how microdomain localization in the plasma membrane affects the function of these complexes is unknown. We aim to study the coupling between LTCC and β adrenergic receptors in different cardiomyocyte microdomains, the distinct involvement of PKA and CAMKII (Ca2+/calmodulin-dependent protein kinase II) and explore how this functional complex is disrupted in heart failure. Methods: Global signaling between LTCCs and β adrenergic receptors was assessed with whole-cell current recordings and western blot analysis. Super-resolution scanning patch-clamp was used to explore the local coupling between single LTCCs and β1AR or β2AR in different membrane microdomains in control and failing cardiomyocytes. Results: LTCC open probability (Po) showed an increase from 0.054±0.003 to 0.092±0.008 when β2AR was locally stimulated in the proximity of the channel (<350 nm) in the transverse tubule microdomain. In failing cardiomyocytes, from both rodents and humans, this transverse tubule coupling between LTCC and β2AR was lost. Interestingly, local stimulation of β1AR did not elicit any change in the Po of LTCCs, indicating a lack of proximal functional interaction between the two, but we confirmed a general activation of LTCC via β1AR. By using blockers of PKA and CaMKII and a Caveolin-3-knockout mouse model, we conclude that the β2AR-LTCC regulation requires the presence of caveolin-3 and the activation of the CaMKII pathway. By contrast, at a cellular "global"level PKA plays a major role downstream β1AR and results in an increase in LTCC current. Conclusions: Regulation of the LTCC activity by proximity coupling mechanisms occurs only via β2AR, but not β1AR. This may explain how β2ARs tune the response of LTCCs to adrenergic stimulation in healthy conditions. This coupling is lost in heart failure; restoring it could improve the adrenergic response of failing cardiomyocytes.

AB - Background: Beta-2 adrenergic receptors (β2ARs) but not beta-2 adrenergic receptors (β1ARs) form a functional complex with L-type Ca2+ channels (LTCCs) on the cardiomyocyte membrane. However, how microdomain localization in the plasma membrane affects the function of these complexes is unknown. We aim to study the coupling between LTCC and β adrenergic receptors in different cardiomyocyte microdomains, the distinct involvement of PKA and CAMKII (Ca2+/calmodulin-dependent protein kinase II) and explore how this functional complex is disrupted in heart failure. Methods: Global signaling between LTCCs and β adrenergic receptors was assessed with whole-cell current recordings and western blot analysis. Super-resolution scanning patch-clamp was used to explore the local coupling between single LTCCs and β1AR or β2AR in different membrane microdomains in control and failing cardiomyocytes. Results: LTCC open probability (Po) showed an increase from 0.054±0.003 to 0.092±0.008 when β2AR was locally stimulated in the proximity of the channel (<350 nm) in the transverse tubule microdomain. In failing cardiomyocytes, from both rodents and humans, this transverse tubule coupling between LTCC and β2AR was lost. Interestingly, local stimulation of β1AR did not elicit any change in the Po of LTCCs, indicating a lack of proximal functional interaction between the two, but we confirmed a general activation of LTCC via β1AR. By using blockers of PKA and CaMKII and a Caveolin-3-knockout mouse model, we conclude that the β2AR-LTCC regulation requires the presence of caveolin-3 and the activation of the CaMKII pathway. By contrast, at a cellular "global"level PKA plays a major role downstream β1AR and results in an increase in LTCC current. Conclusions: Regulation of the LTCC activity by proximity coupling mechanisms occurs only via β2AR, but not β1AR. This may explain how β2ARs tune the response of LTCCs to adrenergic stimulation in healthy conditions. This coupling is lost in heart failure; restoring it could improve the adrenergic response of failing cardiomyocytes.

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U2 - 10.1161/CIRCRESAHA.123.322508

DO - 10.1161/CIRCRESAHA.123.322508

M3 - Article

SN - 0009-7330

VL - 133

SP - 120

EP - 137

JO - Circulation Research

JF - Circulation Research

IS - 2

ER -

Functional LTCC-β2AR Complex Needs Caveolin-3 and Is Disrupted in Heart Failure

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