Dejenerasyon sürecindeki kas dokusunda housekeeping genlerin ekspresyon düzeyinin incelenmesi
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Date
2008
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Biyoteknoloji Enstitüsü
Abstract
RT-PCR and quantitative PCR applications are widely used for the assessment of transcriptional response that cells and tissues develop for external stimuli. For samples that have been collected over a time period or under various physiological conditions, RT-PCR technique is employed to observe any gene of interest. A housekeeping gene whose expression is believed to be stable is used to normalize the expression of the gene of interest. A large battery of genes are being used for the normalization of gene expression in short-term studies where tissue architectural changes are not observed. But under extreme conditions such as the chronic muscle degeneration, the tissue architecture is deteriorated and the cell population exhibit important variations. Thus the gene of choice for normalization has to be carefully assessed under such conditions. The aim of this study is to assess and compare the expression variations of various housekeeping genes on the course of two diverse models of muscle degeneration where large tissue architectural changes are observed. Denervation and tenotomy are the models that are chosen for the study where experimental interventions have been made on the right extremity of rats and the contralateral-side muscles have been utilized as control. The expression of B-actin, TBP, GAPDH and HPRT have been investigated. In order to assess the amount of tissue deterioration, a structural gene of muscle contractile apparatus, the myosin heavy chain is also analyzed.
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normalizasyon, kas dejenerasyonu, housekeeping gen