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Functional constituents of a local serotonergic system, intrinsic to the human coronary artery smooth muscle cells

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Abstract

Human coronary artery smooth muscle cells (HCASMCs) play an important role in the pathogenesis of coronary atherosclerosis and coronary artery diseases (CAD). Serotonin is a mediator known to produce vascular smooth muscle cell mitogenesis and contribute to coronary atherosclerosis. We hypothesize that the HCASMC possesses certain functional constituents of the serotonergic system such as: tryptophan hydroxylase and serotonin transporter. Our aim was to examine the presence of functional tryptophan hydroxylase-1 (TPH1) and serotonin transporter (SERT) in HCASMCs. The mRNA transcripts by qPCR and protein expression by Western blot of TPH1 and SERT were examined. The specificity and accuracy of the primers were verified using DNA gel electrophoresis and sequencing of qPCR products. The functionality of SERT was examined using a fluorescence dye-based serotonin transporter assay. The enzymatic activity of TPH was evaluated using UPLC. The HCASMCs expressed both mRNA transcripts and protein of SERT and TPH. The qPCR showed a single melt curve peak for both transcripts and in sequence analysis the amplicons were aligned with the respective genes. SERT and TPH enzymatic activity was present in the HCASMCs. Taken together, both TPH and SERT are functionally expressed in HCASMCs. These findings are novel and represent an initial step in examining the clinical relevance of the serotonergic system in HCASMCs and its role in the pathogenesis of coronary atherosclerosis and CAD.

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Acknowledgments

The authors thank Dr. Chandra Boosani for his inputs and suggestions on DNA electrophoresis and gene sequencing methodology. The authors thank Dr. Amit N Pandya for the help in conducting UPLC. The authors also thank Dr. Richard Blakely of Vanderbilt University, who generously provided us the HEK-hSERT cells. This work was supported by research grants R01HL090580, R01HL104516 and R01HL112597 from the National Institutes of Health, USA to DK Agrawal. The content of this article is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

Conflict of interest

The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

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    Authors and Affiliations

    1. Center for Clinical and Translational Science, Creighton University School of Medicine, CRISS II Room 510, 2500 California Plaza, Omaha, NE, 68178, USA

      Kannan Baskar & Devendra K. Agrawal

    2. Department of Biomedical Science, Creighton University School of Medicine, Omaha, NE, USA

      Swastika Sur & Devendra K. Agrawal

    3. Department of Psychiatry, Creighton University School of Medicine, Omaha, NE, USA

      Vithyalakshmi Selvaraj

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    1. Kannan Baskar
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    Correspondence to Devendra K. Agrawal.

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    Kannan Baskar and Swastika Sur have contributed equally to this work.

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    Baskar, K., Sur, S., Selvaraj, V. et al. Functional constituents of a local serotonergic system, intrinsic to the human coronary artery smooth muscle cells. Mol Biol Rep 42, 1295–1307 (2015). https://doi.org/10.1007/s11033-015-3874-x

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