Influence of hyaluronic acid synthesis and cumulus mucification on bovine oocyte in vitro maturation, fertilisation and embryo development
Cynthia Gutnisky A , Gabriel C. Dalvit A , Laura N. Pintos A , Jeremy G. Thompson B , Martha T. Beconi A and Pablo D. Cetica A CA Area of Biochemistry, School of Veterinary Sciences, University of Buenos Aires, Chorroarín 280, Buenos Aires C1427CWO, Argentina.
B Research Centre for Reproductive Health, School of Paediatrics and Reproductive Health, The University of Adelaide, 2nd Floor, Medical School South, Adelaide, SA 5005, Australia.
C Corresponding author. Email: pcetica@fvet.uba.ar
Reproduction, Fertility and Development 19(3) 488-497 https://doi.org/10.1071/RD06134
Submitted: 3 October 2006 Accepted: 2 January 2007 Published: 19 March 2007
Abstract
During cumulus–oocyte complex (COC) maturation, cumulus expansion involves the deposition of mucoelastic compounds, especially hyaluronic acid, synthesised from glucose via the hexosamine biosynthesis pathway. The aim of the present study was to determine the effects of uridine monophosphate (UMP) and 6-diazo-5-oxo-l-norleucine (DON), inhibitors of hyaluronic acid synthesis, during bovine oocyte in vitro maturation (IVM) on cumulus expansion, glucose uptake, protein synthesis, cumulus cell number, meiotic maturation, cleavage rate and subsequent embryo development. A further aim of the study was to examine the effect of hyaluronic acid on sperm capacitation and acrosome reaction in relation to the capacity of COCs to be fertilised in vitro. A low correlation between glucose uptake and degree of cumulus expansion was observed. Total and partial inhibition of cumulus expansion was observed with DON and UMP, respectively, and was accompanied by a decrease in glucose uptake with DON. Total protein content and cumulus cell number per COC increased during IVM, but was unaffected by the presence of DON or UMP, as was oocyte meiotic maturation. Rates of cleavage and blastocyst development decreased in oocytes matured with DON and UMP, although this inhibition was reversed when the in vitro fertilisation (IVF) medium contained heparin. Hyaluronic acid induced capacitation and the acrosome reaction, and in IVF medium prevented the inhibition of cleavage and blastocyst development by DON in a similar fashion to heparin. Hyaluronic acid synthesis during cumulus mucification contributes to the penetration and fertilisation of bovine oocytes, most likely by facilitating the processes of capacitation and acrosome reaction. Mucification during IVM is independent of cumulus cell proliferation, COC protein content, oocyte meiotic maturation and subsequent developmental competence once fertilised.
Acknowledgements
This work was supported by a grant from the University of Buenos Aires. The authors thank the Japanese International Cooperation Agency (JICA) for technology transfer and equipment, Deltacar abattoir for ovaries, Astra Laboratories for ultra-pure water and S. Morado and M. Tellado for their technical assistance.
Ali, A. , and Sirard, M. A. (2002). Effect of the absence or presence of various protein supplements on further development of bovine oocyte during in vitro maturation. Biol. Reprod. 66, 901–905.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Hallap, T. , Haard, M. , Jaakma, U. , Larsson, B. , and Rodriguez-Martinez, H. (2004). Does cleansing of frozen–thawed bull semen before assessment provide samples that relate better to potential fertility? Theriogenology 62, 702–713.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Park, J. Y. , Su, Y. Q. , Ariga, M. , Law, E. , Jin, S. L. , and Conti, M. (2004). EGF-like growth factors as mediators of LH action in the ovulatory follicle. Science 303, 682–684.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Salustri, A. , Yanagishita, M. , and Hascall, V. C. (1989). Synthesis and accumulation of hyaluronic acid and proteoglycans in the mouse cumulus cell–oocyte complex during follicle-stimulating hormone-induced mucification. J. Biol. Chem. 264, 13 840–13 847.
| PubMed |
