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17ß-Estradiol regulates expression of K_ATP channels in heart-derived H9c2 cells

Harri J. Ranki, PhD^*, Grant R. Budas, BSc^*, Russell M. Crawford, PhD^*, Anthony M. Davies, BSc^* and Aleksandar Jovanovi, MD, PhD^*,*

^* Tayside Institute of Child Health, Ninewells Hospital & Medical School, University of Dundee, Dundee, Scotland, United Kingdom

View larger version (46K): [in a new window]   Figure 1 Kir6.2 and sulfonylurea receptor 2A (SUR2A) messenger ribonucleic acid levels in untreated and 17ß-estradiol (E2)-treated H9c2 heart cells. (A to D) Reverse transcription-polymerase chain reaction (RT-PCR) products obtained with Kir6.2-, SUR2A- and glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-specific primers from untreated and E2-treated cells. (A1 to C1) Graphs corresponding to RT-PCR products depicted in A to D. Each bar represents mean ± SEM. *p < 0.05 when compared with the control.

View larger version (34K): [in a new window]   Figure 2 Adenosine triphosphate-sensitive potassium channel levels in plasma membrane of untreated and 17-ßestradiol (E2)-treated H9c2 cells. Western blot with anti-Kir6.2 (A) and anti-sulfonylurea receptor 2A (SUR2A) (B) of anti-SUR2A (A) and anti-Kir6.2 (B) immunoprecipitate pellets from membrane fractions from untreated and E2-treated H9c2 cells. (A1 and B1) Graphs correspond to A and B. Each bar represents mean ± SEM. *p < 0.05.IP = immunoprecipitate.

View larger version (60K): [in a new window]   Figure 3 Hypoxia/reoxygenation (H/R)-induced Ca2+ loading in untreated and 17-ßestradiol (E2)-treated H9c2 cells in the absence and presence of HMR 1098 (3 µM) and 5-hydroxydecanoate (5-HD) (50 µM), sarcolemmal and mitochondrial Adenosine triphosphate-sensitive potassium channel blockers, respectively. (A) Epifluorescent digital images of untreated and E2-treated Fura-2 loaded cells before (left panel) and after H/R (right panel). White bar corresponds to 30 µm. Graphs are time courses of intracellular concentration of Ca2+ and correspond to experiments depicted on the left. Each line on graphs represents a single cell from the corresponding image field. (B) Percentage of cells that responded (defined as 50% increase in resting Ca2+ with the time-course pattern of Ca2+ increase as depicted in A)/did not respond to H/R with Ca2+ loading under control conditions (untreated) and 24 h treatment with E2 in the absence (E2) and presence of HMR 1098 (E2 + HMR 1098) and 5-HD (E2 + 5-HD), n = 29 to 45.