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|Title:||Differences in the participation of TGFB superfamily signalling pathways mediating porcine and murine cumulus cell expansion|
|Citation:||Reproduction, 2011; 142(5):647-657|
|Publisher:||Bio Scientifica Ltd|
|Robert B Gilchrist and Lesley J Ritter|
|Abstract:||It is widely held that mammalian cumulus cell (CC) expansion requires oocyte-paracrine signalling, however in three of the four species studied to date, CC expansion occurs in the absence of the oocyte. This study was conducted to examine the paracrine and SMAD/MAPK intracellular signalling mechanism mediating porcine CC expansion, and to compare these to the mouse. Cumulus–oocyte complexes (COCs) and oocyte-free complexes (OOXs) from pigs and eCG-primed mice were treated in vitro with FSH and a broad range of TGFB superfamily antagonists. Expansion of porcine COCs and OOXs was unaffected by neutralisation of growth differentiation factor 9, TGFB, activin A, activin B and a broad spectrum bone morphogenetic protein antagonist. A SMAD-responsive luciferase reporter assay confirmed that porcine oocytes secreted factors that activate SMAD3 and SMAD1/5/8 in granulosa cells, but murine oocytes activated SMAD3 only. Treatment of COCs with a SMAD2/3 phosphorylation inhibitor (SB431542) partially inhibited porcine CC expansion and expression of TNFAIP6, but ablated murine CC expansion. SB431542 was equally effective at attenuating porcine CC expansion in the presence or absence of the oocyte. By contrast, a SMAD1/5/8 phosphorylation inhibitor (dorsomorphin) had no effect on porcine or murine CC function. Inhibition of ERK1/2 and p38 MAPK signalling pathways prevented porcine COC expansion and expression of most matrix genes examined. The activation of CC SMAD signalling by oocytes, and the requirement of SMAD2/3 signalling for expansion, is notably contrasted in pigs and mice. Nonetheless, porcine CC SMAD2/3 signalling is likely to be needed for optimal matrix formation, possibly by facilitating essential MAPK signals.|
|Keywords:||Oocytes; Cells, Cultured; Animals; Swine; Mice; Transforming Growth Factor beta; Paracrine Communication; Signal Transduction; Cell Proliferation; MAP Kinase Signaling System; Gene Expression; Multigene Family; Female; Smad Proteins; Cumulus Cells|
|Rights:||© 2011 Society for Reproduction and Fertility|
|Appears in Collections:||Obstetrics and Gynaecology publications|
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