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Journal of Endocrinology (2006) 190, 85-97    DOI: 10.1677/joe.1.06828
© 2006 Society for Endocrinology

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AMP-activated protein kinase activation modulates progesterone secretion in granulosa cells from hen preovulatory follicles

Lucie Tosca, Sabine Crochet1, Pascal Ferré2, Fabienne Foufelle2, Sophie Tesseraud1 and Joëlle Dupont

Unité de Physiologie de la Reproduction et des Comportements and
1 Station de Recherches Avicoles, Institut National de la Recherche Agronomique, 37380 Nouzilly, France
2 INSERM UMR S671, Centre Biomédical des Cordeliers, Université Pierre et Marie Curie-Paris 6, Paris F-75006, France

(Requests for offprints should be addressed to J Dupont; Email: jdupont{at}tours.inra.fr)

AMP-activated protein kinase (AMPK) is a fuel sensor in glucose, lipid, and cholesterol metabolism. Using RT-PCR and Western blot, AMPK subunits mRNAs ({alpha}1/2, ß1/2, and {gamma}1/2) and proteins ({alpha}1/2 and ß1/2) can be found in the hen preovulatory follicles and precisely in both granulosa and theca cells. These preovulatory follicles are organized in a hierarchy according to their size (F5/6 to F1). The smallest number (F1) corresponds to the largest size and the latest mature stage. Phosphorylation of AMPK{alpha} on Thr172 and of acetyl-CoA carboxylase on Ser79 are higher in F4 and F3 than in F1 granulosa cells. However, they are not affected in F4–F1 theca cells. Treatment with 1 mM 5-amino-imidazole-4-carboxyamide-1-ß-D-ribofuranoside (AICAR), an activator of AMPK, dose dependently increased phosphorylation of AMPK{alpha} on Thr172 in primary F3/4 and F1 granulosa cells. In the absence of FSH, AICAR treatment increased progesterone, P450 side chain cleavage and steroidogenic acute regulatory (StAR) production in both F3/4 and F1 granulosa cells. However, in the presence of FSH, AICAR treatment for 36 h increased progesterone secretion, StAR protein levels and reduced extracellular signal-regulated kinase (ERK)1/2 phosphorylation in F3/4 granulosa cells. Opposite data were observed in F1 granulosa cells. Adenovirus-mediated expression of dominant-negative AMPK totally restored the effects of AICAR on FSH-induced progesterone secretion, StAR protein production, and ERK1/2 phosphorylation in F3/4 and F1 granulosa cells. Using a specific inhibitor of ERK1/2 (U0126), we also showed that this kinase is a negative regulator of the FSH-induced progesterone secretion in F3/4 and F1 granulosa cells, suggesting that AICAR-mediated AMPK activation modifies FSH-induced progesterone secretion differently through the ERK1/2 signaling pathway in hen F3/4 and F1 granulosa cells.




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