Store-operated calcium entry is present in HL-1 cardiomyocytes and contributes to resting calcium

Chad D. Touchberry; Chris J. Elmore; Tien M. Nguyen; Jon J. Andresen; Xiaoli Zhao; Matthew Orange; Noah Weisleder; Marco Brotto; William C. Claycomb; Michael J. Wacker

doi:10.1016/j.bbrc.2011.10.133

Store-operated calcium entry is present in HL-1 cardiomyocytes and contributes to resting calcium

Chad D. Touchberry, Chris J. Elmore, Tien M. Nguyen, Jon J. Andresen, Xiaoli Zhao, Matthew Orange, Noah Weisleder, Marco Brotto, William C. Claycomb, Michael J. Wacker

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

47 Scopus citations

Abstract

Store-operated Ca ²⁺ entry (SOCE) has recently been shown to be of physiological and pathological importance in the heart, particularly during cardiac hypertrophy. However, measuring changes in intracellular Ca ²⁺ during SOCE is very difficult to study in adult primary cardiomyocytes. As a result there is a need for a stable and reliable in vitro model of SOCE which can be used to test cardiac drugs and investigate the role of SOCE in cardiac pathology. HL-1 cells are the only immortal cardiomyocyte cell line available that continuously divides and spontaneously contracts while maintaining phenotypic characteristics of the adult cardiomyocyte. To date the role of SOCE has not yet been investigated in the HL-1 cardiac cell line. We report for the first time that these cells expressed stromal interaction molecule 1 (STIM1) and the Ca ²⁺ release-activated Ca ²⁺ (CRAC) channel Orai1, which are essential components of the SOCE machinery. In addition, SOCE was tightly coupled to sarcoplasmic reticulum (SR)-Ca ²⁺ release in HL-1 cells, and such response was not impaired in the presence of voltage dependent Ca ²⁺ channels (L-type and T-type channels) or reverse mode Na ⁺/Ca ²⁺ exchanger (NCX) inhibitors. We were able to abolish the SOCE response with known SOCE inhibitors (BTP-2 and SKF-96365) and by targeted knockdown of Orai1 with RNAi. In addition, knockdown of Orai1 resulted in lower baseline Ca ²⁺ and an attenuated response to thapsigargin (TG) and caffeine, indicating that SOCE may play a role in Ca ²⁺ homeostasis during unstressed conditions in cardiomyocytes. Currently, there is little knowledge about SOCE in cardiomyocytes, and the present results suggest that HL-1 cells will be of great utility in investigating the role of SOCE in the heart.

Original language	English
Pages (from-to)	45-50
Number of pages	6
Journal	Biochemical and Biophysical Research Communications
Volume	416
Issue number	1-2
DOIs	https://doi.org/10.1016/j.bbrc.2011.10.133
State	Published - Dec 9 2011

Funding

This research was supported by grants from the American Heart Association to MW (11SDG5330016), MB (0535555N), XZ (10SDG2630086), and JA (0735053N). NW was supported by NIH/NIAMS (AR054793). WC was supported by grants from NIH (HL076498) and the Joe W. and Dorothy Brown Foundation.

Funders	Funder number
NIH NIAMS	AR054793
National Institutes of Health (NIH)	HL076498
National Institute of Arthritis and Musculoskeletal and Skin Diseases	R00AR054793
American the American Heart Association	10SDG2630086, 11SDG5330016, 0535555N, 0735053N
Joe W. and Dorothy Dorsett Brown Foundation

Keywords

Cardiomyocytes
HL-1 cells
Orai
Stim
Store-operated calcium entry

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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@article{56bcc98769d243bd837f665f63b98aac,

title = "Store-operated calcium entry is present in HL-1 cardiomyocytes and contributes to resting calcium",

abstract = "Store-operated Ca 2+ entry (SOCE) has recently been shown to be of physiological and pathological importance in the heart, particularly during cardiac hypertrophy. However, measuring changes in intracellular Ca 2+ during SOCE is very difficult to study in adult primary cardiomyocytes. As a result there is a need for a stable and reliable in vitro model of SOCE which can be used to test cardiac drugs and investigate the role of SOCE in cardiac pathology. HL-1 cells are the only immortal cardiomyocyte cell line available that continuously divides and spontaneously contracts while maintaining phenotypic characteristics of the adult cardiomyocyte. To date the role of SOCE has not yet been investigated in the HL-1 cardiac cell line. We report for the first time that these cells expressed stromal interaction molecule 1 (STIM1) and the Ca 2+ release-activated Ca 2+ (CRAC) channel Orai1, which are essential components of the SOCE machinery. In addition, SOCE was tightly coupled to sarcoplasmic reticulum (SR)-Ca 2+ release in HL-1 cells, and such response was not impaired in the presence of voltage dependent Ca 2+ channels (L-type and T-type channels) or reverse mode Na +/Ca 2+ exchanger (NCX) inhibitors. We were able to abolish the SOCE response with known SOCE inhibitors (BTP-2 and SKF-96365) and by targeted knockdown of Orai1 with RNAi. In addition, knockdown of Orai1 resulted in lower baseline Ca 2+ and an attenuated response to thapsigargin (TG) and caffeine, indicating that SOCE may play a role in Ca 2+ homeostasis during unstressed conditions in cardiomyocytes. Currently, there is little knowledge about SOCE in cardiomyocytes, and the present results suggest that HL-1 cells will be of great utility in investigating the role of SOCE in the heart.",

keywords = "Cardiomyocytes, HL-1 cells, Orai, Stim, Store-operated calcium entry",

author = "Touchberry, \{Chad D.\} and Elmore, \{Chris J.\} and Nguyen, \{Tien M.\} and Andresen, \{Jon J.\} and Xiaoli Zhao and Matthew Orange and Noah Weisleder and Marco Brotto and Claycomb, \{William C.\} and Wacker, \{Michael J.\}",

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T1 - Store-operated calcium entry is present in HL-1 cardiomyocytes and contributes to resting calcium

AU - Touchberry, Chad D.

AU - Elmore, Chris J.

AU - Nguyen, Tien M.

AU - Andresen, Jon J.

AU - Zhao, Xiaoli

AU - Orange, Matthew

AU - Weisleder, Noah

AU - Brotto, Marco

AU - Claycomb, William C.

AU - Wacker, Michael J.

PY - 2011/12/9

Y1 - 2011/12/9

N2 - Store-operated Ca 2+ entry (SOCE) has recently been shown to be of physiological and pathological importance in the heart, particularly during cardiac hypertrophy. However, measuring changes in intracellular Ca 2+ during SOCE is very difficult to study in adult primary cardiomyocytes. As a result there is a need for a stable and reliable in vitro model of SOCE which can be used to test cardiac drugs and investigate the role of SOCE in cardiac pathology. HL-1 cells are the only immortal cardiomyocyte cell line available that continuously divides and spontaneously contracts while maintaining phenotypic characteristics of the adult cardiomyocyte. To date the role of SOCE has not yet been investigated in the HL-1 cardiac cell line. We report for the first time that these cells expressed stromal interaction molecule 1 (STIM1) and the Ca 2+ release-activated Ca 2+ (CRAC) channel Orai1, which are essential components of the SOCE machinery. In addition, SOCE was tightly coupled to sarcoplasmic reticulum (SR)-Ca 2+ release in HL-1 cells, and such response was not impaired in the presence of voltage dependent Ca 2+ channels (L-type and T-type channels) or reverse mode Na +/Ca 2+ exchanger (NCX) inhibitors. We were able to abolish the SOCE response with known SOCE inhibitors (BTP-2 and SKF-96365) and by targeted knockdown of Orai1 with RNAi. In addition, knockdown of Orai1 resulted in lower baseline Ca 2+ and an attenuated response to thapsigargin (TG) and caffeine, indicating that SOCE may play a role in Ca 2+ homeostasis during unstressed conditions in cardiomyocytes. Currently, there is little knowledge about SOCE in cardiomyocytes, and the present results suggest that HL-1 cells will be of great utility in investigating the role of SOCE in the heart.

AB - Store-operated Ca 2+ entry (SOCE) has recently been shown to be of physiological and pathological importance in the heart, particularly during cardiac hypertrophy. However, measuring changes in intracellular Ca 2+ during SOCE is very difficult to study in adult primary cardiomyocytes. As a result there is a need for a stable and reliable in vitro model of SOCE which can be used to test cardiac drugs and investigate the role of SOCE in cardiac pathology. HL-1 cells are the only immortal cardiomyocyte cell line available that continuously divides and spontaneously contracts while maintaining phenotypic characteristics of the adult cardiomyocyte. To date the role of SOCE has not yet been investigated in the HL-1 cardiac cell line. We report for the first time that these cells expressed stromal interaction molecule 1 (STIM1) and the Ca 2+ release-activated Ca 2+ (CRAC) channel Orai1, which are essential components of the SOCE machinery. In addition, SOCE was tightly coupled to sarcoplasmic reticulum (SR)-Ca 2+ release in HL-1 cells, and such response was not impaired in the presence of voltage dependent Ca 2+ channels (L-type and T-type channels) or reverse mode Na +/Ca 2+ exchanger (NCX) inhibitors. We were able to abolish the SOCE response with known SOCE inhibitors (BTP-2 and SKF-96365) and by targeted knockdown of Orai1 with RNAi. In addition, knockdown of Orai1 resulted in lower baseline Ca 2+ and an attenuated response to thapsigargin (TG) and caffeine, indicating that SOCE may play a role in Ca 2+ homeostasis during unstressed conditions in cardiomyocytes. Currently, there is little knowledge about SOCE in cardiomyocytes, and the present results suggest that HL-1 cells will be of great utility in investigating the role of SOCE in the heart.

KW - Cardiomyocytes

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KW - Orai

KW - Stim

KW - Store-operated calcium entry

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Store-operated calcium entry is present in HL-1 cardiomyocytes and contributes to resting calcium

Abstract

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Keywords

ASJC Scopus subject areas

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