Show simple item record

dc.contributor.advisorSelvaraj, Vidhyaen_US
dc.contributor.authorBaskar, Kannanen_US
dc.date.accessioned2013-12-18T15:33:54Z
dc.date.available2013-12-18T15:33:54Z
dc.date.issued2013-11en_US
dc.identifier.urihttp://hdl.handle.net/10504/47427
dc.description.abstractAim: Human coronary artery smooth muscle cell (HCASMC) plays 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 (VSMC) mitogenesis and contribute to coronary atherosclerosis. We hypothesize that the human coronary artery smooth muscle cell possesses certain vital elements of the serotonergic system, viz. tryptophan hydroxylase and serotonin transporter, which play a critical role in the synthesis and VSMC-mitogenic action of serotonin, respectively. Our aim was to demonstrate the presence of functional tryptophan hydroxylase 1 and serotonin transporter in HCASMC.|Methods: HCASMCs from passages 3 -6 were used for the experiments. The messenger ribonucleic acid (mRNA) transcripts and protein expression of tryptophan hydroxylase1 (TPH1) and serotonin transporter (SERT) were examined by real-time PCR (Real time polymerase chain reaction) and western blot analysis, respectively. The specificity and accuracy of the primers was verified using DNA (Deoxyribonucleic acid) gel electrophoresis and sequencing of real-time PCR products. The functionality of SERT was examined using a fluorescence dye based serotonin transporter assay and the enzymatic activity of TPH was demonstrated using Ultra Performance Liquid Chromatography (UPLC). |Results: mRNA transcripts and protein of both SERT and TPH are expressed in HCASMCs. The real-time PCR showed a single melt curve peak for both transcripts, and the amplicons when sequenced aligned with the respective genes. The Western blots revealed distinct bands for SERT and TPH at 70 KDa and 50 KDa, respectively. Serotonin transporter activity was present in HCASMCs. UPLC of the HCASMC lysate, to which the substrates and catalysts for the TPH enzyme had been provided, revealed the presence of 5-hydroxy tryptophan. Conclusion: Both tryptophan hydroxylase and serotonin transporter are expressed in HCASMCs. The expressed proteins are functional. These findings are novel and an initial step in examining the clinical relevance of the serotonergic system in HCASMC and its role in the pathogenesis of coronary atherosclerosis and CADen_US
dc.language.isoen_USen_US
dc.publisherCreighton Universityen_US
dc.rightsCopyright is retained by the Author. A non-exclusive distribution right is granted to Creighton University and to ProQuest following the publishing model selected above.en_US
dc.subject.meshSerotonin--physiologyen_US
dc.subject.meshCoronary Artery Disease--physiopathologyen_US
dc.subject.meshMuscle, Smooth, Vascularen_US
dc.titleFunctional constituents of a local serotonergic system, intrinsic to the human coronary artery smooth muscle cellen_US
dc.typeThesis
dc.rights.holderKannan Baskaren_US
dc.publisher.locationOmaha, Nebraskaen_US
dc.description.noteProQuest Traditional Publishing Optionen_US
dc.description.pagesxxii, 168 pagesen_US
dc.contributor.cuauthorBaskar, Kannanen_US
dc.degree.levelMS (Master of Science)en_US
dc.degree.disciplineClinical and Translational Science (graduate program)en_US
dc.degree.nameM.S. in Clinical and Translational Scienceen_US
dc.degree.grantorGraduate Schoolen_US
dc.degree.committeeAgrawal, Devendraen_US
dc.degree.committeeDrescher, Kristenen_US
dc.degree.committeePatterson, Ericen_US


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record