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GABAB receptors are expressed in human aortic smooth muscle cells and regulate the intracellular Ca2+ concentration

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Abstract

The aim of this study was to investigate the expression of GABAB receptors, a subclass of receptors to the inhibitory neurotransmitter gamma-aminobutyric acid (GABAB), in human aortic smooth muscle cells (HASMCs), and to explore if altering receptor activation modified intracellular Ca2+ concentration ([Ca2+]i) of HASMCs. Real-time PCR, western blots and immunofluorescence were used to determine the expression of GABABR1 and GABABR2 in cultured HASMCs. Immunohistochemistry was used to localize the two subunits in human left anterior descending artery (LAD). The effects of the GABAB receptor agonist baclofen on [Ca2+]i in cultured HASMCs were demonstrated using fluo-3. Both GABABR1 and GABABR2 mRNA and protein were identified in cultured HASMCs and antibody staining was also localized to smooth muscle cells of human LAD. 100 μM baclofen caused a transient increase of [Ca2+]i in cultured HASMCs regardless of whether Ca2+ was added to the medium, and the effects were inhibited by pre-treatment with CGP46381 (selective GABAB receptor antagonist), pertussis toxin (a Gi/o protein inhibitor), and U73122 (a phospholipase C blocker). GABAB receptors are expressed in HASMCs and regulate the [Ca2+]i via a Gi/o-coupled receptor pathway and a phospholipase C activation pathway.

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Abbreviations

GABA:

γ-Aminobutyric acid

GABABR1 :

γ-Aminobutyric acid type B receptor subunit one

GABABR2 :

γ-Aminobutyric acid type B receptor subunit two

HASMCs:

Human aortic smooth muscle cells

VSMCs:

Vascular smooth muscle cells

PCR:

Polymerase chain reaction

mRNA:

Messenger RNA

G proteins:

Guanine nucleotide-binding proteins

[Ca2+]i:

Intracellular Ca2+ concentration

SMCM:

Smooth muscle cell medium

LAD:

Left anterior descending artery

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Acknowledgments

This work was supported by grant from National Natural Science Foundation of China (No. 81271038), grant from the Department of Science and Technology of Shandong Province of China (No. ZR2012HM022) to ZYC, grant from National Nature Science Foundation of China (No. 80200212) and grant from the Department of Science and Technology of Shandong Province of China (No. ZR2012HQ029) to XPW.

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The authors have no conflict of interests to declare.

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Authors and Affiliations

  1. The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Qilu Hospital, Shandong University, Jinan, Shandong, China

    Xu-Ping Wang

  2. Department of Ophthalmology, Qilu Hospital, Shandong University, 107 Wenhuaxi Road, Jinan, 250012, Shandong, China

    Zhen-Ying Cheng

  3. School of Optometry and Vision Science, Faculty of Health and Vision Improvement Domain, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia

    Katrina L. Schmid

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  1. Xu-Ping Wang
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Correspondence to Zhen-Ying Cheng.

Additional information

X.-P. Wang, Z.-Y. Cheng made the same contribution to this paper.

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Wang, XP., Cheng, ZY. & Schmid, K.L. GABAB receptors are expressed in human aortic smooth muscle cells and regulate the intracellular Ca2+ concentration. Heart Vessels 30, 249–257 (2015). https://doi.org/10.1007/s00380-014-0499-2

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  • DOI: https://doi.org/10.1007/s00380-014-0499-2

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