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(379) REAL TIME RT-PCR ANALYSIS OF GENES EXPRESSED BY HUMAN GRANULOSA CELLS IN A VARIETY OF INFERTILE DISORDERS. Quinn, Michael1, Stanton, Jo-Ann1, Green, David 1, 1 Department of Anatomy & Structural Biology, Dunedin, New Zealand ABSTRACT- Human granulosa cells play a crucial role in follicle development and are essential for successful oocyte maturation and ovulation. Molecular defects in granulosa cells are likely to play an important role in human female infertility. Human granulosa cells from normal females and females suffering a range of infertility disorders were collected and purified at the time of in vitro fertilization (IVF): normal (or male-factor, n=18); endometriosis (n=2); polycystic ovary syndrome (PCO, n=3); tubal (n=5) and idiopathic (unknown) (n=5). Quantitative mRNA expression levels of 8 candidate genes identified from a SAGE catalogue of human granulosa cells were measured by real time RT-PCR. These genes were: hydroxysteroid (11-beta) dehydrogenase; retinol binding protein 1; SCARB1 receptor; follistatin; follicle stimulating hormone (FSH) receptor; progesterone receptor membrane component 1; progesterone receptor membrane component 2 and decidual protein induced by progesterone. As a comparison, mRNA expression of the same genes in human white blood cells was also measured. Gene expression was normalized to beta-actin (standard curve method). Results were analyzed by 1-way ANOVA and a Tukey follow-up test (5% significance). No significant difference in the level of mRNA expression was detected between the control group and the infertility disorder groups investigated. However, several individual patients from the PCO and idiopathic groups showed over-expression of some genes relative to the normal group. These patients may represent subpopulations in heterogeneous syndromes. The over-expressed genes may therefore be useful markers for the disorders. KEY WORDS: infertility, SAGE, granulosa, real time RT-PCR |
