Development of real-time reverse transcription polymerase chain reaction assays to quantify insulin-like growth factor-1 receptor and insulin receptor expression in equine tissue

dc.contributor.advisorSchulman, Martin L.
dc.contributor.coadvisorGuthrie, Alan John
dc.contributor.coadvisorQuan, Melvyn
dc.contributor.emailshughes.vet@gmail.comen
dc.contributor.postgraduateHughes, Stephen Bernard
dc.date.accessioned2013-09-09T12:06:42Z
dc.date.available2012-08-14en
dc.date.available2013-09-09T12:06:42Z
dc.date.created2012-04-13en
dc.date.issued2011en
dc.date.submitted2012-08-08en
dc.descriptionDissertation (MMedVet)--University of Pretoria, 2011.en
dc.description.abstracthas been significant progress in the development of new technologies and methodologies to characterize gene expression. The fluorescent-based real-time reverse transcription (RT) polymerase chain reaction (PCR) is an important tool used for clinical and molecular research, biotechnology and as a diagnostic test. Insulin-like growth factors (IGF-1 and IGF-2) and insulin are ubiquitously expressed and play important roles in the regulation of cell growth, differentiation and the maintenance of cell differentiation in mammals. The IGF system (IGF-1, IGF-2, IGF -1 receptor, IGF-2 receptor and six IGF-binding proteins) and insulin are consequently essential to most aspects of male and female reproduction. IGF-1 is produced in multiple tissues but predominately in the liver, from where it enters the circulation. Insulin is secreted by β-cells of the pancreas’ islets of Langerhans. Both IGF-1 and insulin polypeptides bind to specific cell surface receptors. These receptors are members of the superfamily known as tyrosine protein kinases, and are composed of two α and two β subunits linked by disulfide bonds to form an αβ–αβ heterotetramer. The α subunits include ligand binding sites, whereas the β subunits contain tyrosine kinase activity. The aim of this project was to develop real-time RT-PCR assays for quantification of equine insulin-like growth factor-1 receptor (IGF-1R) and insulin receptor (INS-R) mRNA. The assays were developed using stallion testicular tissue samples, obtained by excisional biopsy, from three horse breeds (Friesan, Thoroughbred and Warmblood). The assays developed were efficient, sensitive and had a broad linear range of detection (seven logs for IGF-1R and six logs for INS-R). The assays worked well in our hands and were both sensitive and specific for the detection of equine IGF-1R and INS-R mRNA in a variety of equine tissues.en
dc.description.availabilityUnrestricteden
dc.description.departmentProduction Animal Studiesen
dc.identifier.citationHughes, SB 2011, Development of real-time reverse transcription polymerase chain reaction assays to quantify insulin-like growth factor-1 receptor and insulin receptor expression in equine tissue, MMedVet dissertation, University of Pretoria, Pretoria, viewed yymmdd <http://hdl.handle.net/2263/31135>en
dc.identifier.otherE12/4/294/gmen
dc.identifier.upetdurlhttp://upetd.up.ac.za/thesis/available/etd-08082012-144801/en
dc.identifier.urihttp://hdl.handle.net/2263/31135
dc.language.isoenen
dc.publisherUniversity of Pretoriaen_ZA
dc.rights© 2011, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. E12/4/294/en
dc.subjectInsulin receptoren
dc.subjectUCTD
dc.subjectEquine tissue
dc.subjectTranscription polymerase
dc.subjectTyrosine protein kinases
dc.titleDevelopment of real-time reverse transcription polymerase chain reaction assays to quantify insulin-like growth factor-1 receptor and insulin receptor expression in equine tissueen
dc.typeDissertationen

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