WCRB2014 POSTER PRESENTATIONS (1) (335 abstracts)
Pinacidil, a KATP channel opening drug, protects human oocytes against metabolic stress
Gonçalo M T Fernandes 1 , Mandy Gall 2 , Ellen Drew 2 , Evelyn Barratt 2 , Oladipo Madamidola 3 , Sean G Brown 3 , Alison M Milne 1 , Sarah Martins da Silva 1 , Christopher Barratt 1 & Aleksandar Jovanović 1
1University of Dundee, Dundee, UK; 2Ninewells Hospital, Dundee, UK; 3Abertay University, Dundee, UK.
Introduction: The primary treatment of infertility is assisted reproductive technology (ART). However, removal of an oocyte from its natural environment and exposure to in vitro conditions diminishes oocyte quality during the process of ART. In our preliminary studies we have shown that oocytes express cytoprotective ATP-sensitive K+ (KATP) channels. Therefore, the aim of our study was to elucidate whether activation of these channels protects human oocytes against metabolic stress.
Materials and methods: Supernumerary human oocytes in different stages of maturation were obtained from patients undergoing IVF and ICSI procedures. Cells were stained with Ca2+-sensitive dye Fluo-3 and continuously monitored for 120 min using laser confocal microscopy in the presence and absence of pinacidil, a KATP channel opener, and 2,4 dinitrophenole (DNP), an inhibitor of oxidative phosphorylation.
Results and discussion: We have found that intracellular Ca2+, an indicator of the metabolic state of cells, increases over time when oocytes are left incubating in vitro showing that in vitro conditions are stressful for oocytes. Pinacidil (100 μM) significantly decreased the rate of Ca2+ increase. To induce more severe metabolic stress, we exposed oocytes to DNP (100 μM). DNP induced significant increase in intracellular Ca2+. This effect of DNP was inhibited by pinacidil (100 μM) suggesting that the activation of KATP channels protects oocytes against stress. Consequently, a KATP channels-based strategy that would reduce cellular stress could have a potential to significantly improve the success rate of assisted conception by improving the quality of oocytes used for this purpose.
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ISSN 2052-1472 (online)
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