SFEBES2016 Poster Presentations Reproduction (33 abstracts)
Insight into the molecular mechanisms underlying alterations in gonadotropin receptor activity in polycystic ovarian syndrome
Lisa Owens , Avi Lerner , Silvia Sposini , George Christopoulos , Shiran Khanjani , Razia Islam , Stuart Lavery , Vicky Tsui , Kate Hardy , Stephen Franks & Aylin Hanyaloglu
Imperial College London, London, UK.
Introduction: Polycystic ovary syndrome (PCOS) is a common endocrine disorder, affecting 5–10% of women of reproductive age, and is the major cause of anovulatory infertility. Aberrant secretion and/or action of gonadotropins are implicated but, to date, we have only limited knowledge about the precise mechanisms involved. Recent genome wide association studies have discovered signals at loci close to the genes coding for gonadotropin receptors. The functional significance of these polymorphisms is, as yet, unclear and represents a key area for research.
Methods: In this study granulosa-lutein (GL) cells were obtained from women with and without PCOS undergoing IVF. HEK293 cells were also used as an ovarian PCOS model by stable transfection with FLAG-LHR, transient transfection with HA-FSHR and 24-hour treatment with DHT. RNA was extracted and qPCR performed to analyse differential gene expression. Cyclic AMP production was measured after administration of luteinising hormone (LH) and follicle stimulating hormone (FSH) to cultured cells using a second messenger accumulation assay. Intracellular calcium signalling was measured after administering LH using calcium fluorescent indicators.
Results: Increased expression of full-length FSH (P=0.02) and LH (P=0.05) receptor RNA was seen in PCOS GL cells, along with increased expression of signaling/trafficking molecules β arrestin-2 (P=0.03), PDZ-protein GIPC (P=0.07) and APPL1 (P=0.005). No significant differences were seen in expression of LH receptor splice variants. CyclicAMP level measured after administration of LH for 5 minutes was higher in GL cells from PCOS than from controls (x4 fold). Similarly cAMP produced after administration of LH to HEK cells was higher in cells pre-treated with DHT (x3.5fold). cAMP measured after administration of FSH was however negligible in all groups, suggesting involvement of an alternative to the traditional Gs pathway. Administration of LH activated a calcium signaling response.
Conclusion: These results reveal multiple molecular alterations of LH receptor action and downstream signaling in GL cells from women with PCOS.
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Endocrine Abstracts
ISSN 1470-3947 (print) | ISSN 1479-6848 (online)
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