Role of the hyaluronan receptor CD44 during porcine oocyte maturation
ABSTRACT Previous our studies have shown that CD44, the principal receptor for hyaluronan, is present on cumulus cells during oocyte maturation. Although hyaluronan-CD44 interaction has been implicated in cumulus expansion and/or oocyte maturation, the full significance of CD44 remains unknown. The objective of the present study was to further investigate the role of CD44 in cumulus expansion and oocyte maturation in pigs. We demonstrate here in that CD44 has a key role in oocyte maturation but not in cumulus expansion. Previous studies have reported the physiological significance of cumulus expansion in oocyte maturation. However, our results suggest that cumulus expansion is a necessary condition for oocyte maturation, but that it is not sufficient on its own. Furthermore, western blot analysis demonstrated that the CD44 of the in vitro-matured cumulus-oocyte complexes (COCs) had a larger molecular weight and more terminal sialic acid, which has been proven to inhibit the hyaluronan-binding ability of the receptor, than the CD44 of the in vivo-matured COCs, indicating that the hyaluronan-CD44 interactions during in vitro maturation might be insufficient compared with those in vivo. The insufficient interactions of hyaluronan-CD44 during in vitro maturation may cause the inferior capacity of fertilization and development of oocytes matured in vitro.
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ABSTRACT: The CD44 family belongs to a larger group of hyaluronic acid-binding proteins and plays important roles in oocyte maturation, fertilization and preimplantational embryo development. We analyzed the CD44 receptor in sheep oocytes and embryos. Immature oocytes (N = 66) were obtained from a local abattoir; mature oocytes (N = 35) and embryos (N = 41) were obtained by laparotomy from adult hair ewes submitted to ovarian stimulation treatment. The CD44 mRNA was detected by hemi-nested PCR, after reverse transcription, while proteins were located by indirect immunofluorescence, using anti-human CD44 monoclonal antibody. Human lymphocytes and immature bovine oocytes were used as positive and negative controls, respectively. Assessment of the oocyte nuclear stages as well as classification of the embryonic development stage were made with Hoechst 33342 staining. Indirect immunofluorescence detected CD44 expression on the surface of mature oocytes and embryos; immature oocytes did not take up the stain. These findings were supported by the RT-PCR data, which showed no mRNA templates for CD44, even after two consecutive amplifications, in material from immature oocytes and cumulus cells. The CD44 amplicons were detected after a second hemi-nested PCR in mature oocytes and embryos. The finding of CD44 in mature oocytes and preimplantational embryos could reflect the expression profile of hyaluronic acid during terminal folliculogenesis and preimplantational embryo development in sheep.Genetics and molecular research: GMR 01/2012; 11(2):799-809. DOI:10.4238/2012.April.3.2 · 0.85 Impact Factor
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ABSTRACT: Fertilization and early embryo development relies on a complex interplay between the Fallopian tube and the gametes before and after fertilization. Thereby the oviduct, as a dynamic reproductive organ, enables reception, transport and maturation of male and female gametes, their fusion, and supports early embryo development. This paper reviews current knowledge regarding physiological processes behind the transport of boar spermatozoa, their storage in and release from the functional sperm reservoir (SR), and of the interactions that newly ovulated oocytes play within the tube during their transport to the site of fertilization. Experimental evidence of an ovarian control on sperm release from the SR is highlighted. Furthermore, the impact of oviductal secretion on sperm capacitation, oocyte maturation, fertilization and early embryo development is stressed.Reproduction in Domestic Animals 07/2008; 43 Suppl 2:245-51. DOI:10.1111/j.1439-0531.2008.01169.x · 1.18 Impact Factor
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ABSTRACT: The effects of adding the extracellular glycosaminoglycans (GAGs), hyaluronic acid (HA) and chondroitin sulphate (CS) to rat in vitro fertilisation (IVF) media were assessed. Metaphase II (MII) oocytes were also incubated in GAG-supplemented modified rat 1-cell embryo culture medium (mR1ECM+BSA) for 3 days. Cytoplasmic fragmentation was significantly reduced in mR1ECM+BSA with HA (39.0-48.0%) compared with the control (82.0%). In IVF experiments, neither HA (8.0-30.8%) nor CS (9.7-42.5%) improved fertilisation rates compared with controls fertilised in M16 (47.2%) or enriched Krebs-Ringer bicarbonate solution (61.5%). RT-PCR and Western blot were used to probe for CD44 mRNA and protein in Sprague-Dawley gametes and cumulus cells. CD44 was identified in cumulus cells, suggesting a role for oocyte maturation and cumulus expansion. The CD44 protein was also present on caudal epididymal spermatozoa that were highly stimulated by CS in vitro implicating a role in fertilisation for CS and CD44.Reproduction 04/2008; 135(3):311-9. DOI:10.1530/REP-07-0267