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Vertebrate reproductive science and technology
RESEARCH ARTICLE

224. Disruption of bi-directional oocyte-cumulus paracrine signalling during oocyte in vitro maturation reduces subsequent mouse fetal survival

C. X. Yeo A , R. B. Gilchrist A and M. Lane A B
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A Research Centre For Reproductive Health, University of Adelaide, Adelaide, SA, Australia.

B Repromed, Dulwich, SA, Australia.

Reproduction, Fertility and Development 20(9) 24-24 https://doi.org/10.1071/SRB08Abs224
Published: 28 August 2008

Abstract

During folliculogenesis, oocyte to cumulus cell (CC) bi-directional communication is essential for normal development of the oocyte. We recently showed that addition of recombinant oocyte paracrine factor growth differentiation factor 9 (GDF9) during mouse oocyte in vitro maturation (IVM) increased fetal viability. GD.F. 9 signals through SMAD 2/3. Hence the effects of disrupting SMAD2/3 signalling and its interaction with FSH/EGF during IVM on oocyte development and subsequent fetal outcomes were investigated. Cumulus-oocyte complexes (COCs) from antral follicles (n = 400–500) of eCG treated pre-pubertal (C57BL/6xCBA F1 hybrid) mice were cultured for 18 h in Waymouth's medium+5% serum, with or without 50 mIU/mL FSH and 10ng/mL EGF, SMAD2/3 inhibitor SB-431542 (4µM), or its 0.04% DMSO control. Meiotic maturation was assessed by first polar body (PB1) extrusion immediately after culture. COCs were fertilised and cultured to the blastocyst stage in G1.2/G2.2 media at 37°C in 6%CO2:5%O2:89%N2. Blastocysts were either transferred to pseudo-pregnant Swiss females or differentially stained. Pregnancy outcome was assessed on Day 18 of pregnancy. Inhibition of SMAD 2/3 signalling did not alter meiotic maturation. No differences were observed in the percentage of blastocysts or hatching blastocysts from cleaved embryos with SMAD2/3 inhibition or the absence of FSH/EGF. However, IVM with SB-431542 or without FSH/EGF significantly decreased (P < 0.001) blastocyst inner cell mass percentages (26% v. 35% control;18% v. 28% control respectively). Fetal survival (fetuses per embryo transferred) of oocytes matured with SB-431542 was significantly decreased (30% v. 50% controls; P < 0.05) although implantation rates and subsequent fetal weights were unaffected. These findings demonstrate the importance of oocyte-CC communication throughout IVM. Inhibition of oocyte signalling through SMAD2/3 resulted in reduced blastocyst quality and fetal survival; outcomes similar to that of oocytes matured without FSH/EGF. Oocyte–cumulus cell bi-directional communication is thus an important feature of oocyte viability and has a substantial impact on subsequent fetal outcomes.