CD44 family belongs to a large group of proteins that bind to hyaluronic acid. It has important role in oocyte maturation, fertilization and embryo development. We analyzed CD44 in oocytes and embryos of goat and different breeds of sheep. We used both Kermani and baluchi breeds of sheep, two Rayeni and Tali goats. Domestic animals kept after feeding for 3 months exude embryos and oocyte. After this, the animals were cured by keeping full progesterone for 14 days in the vagina. Gonadotropin-releasing hormone (GnRH) hormone was injected from the animals' vagina and made ready for sampling after 1 day. After that, mature oocyte was exuded from the animals' uterus and they were taken for slaughter. To get the embryos, male animals were kept beside them. After 6 days these animals were taken for slaughter. In another way, ovary collected from the slaughter house exuded immature oocyte. In the laboratory, immature and mature oocyte was produced. So by keeping sperm under environmental condition embryo culture is produced. The samples exude RNA using kit; after that, cDNA was produced by special protocol. The products produced by conventional PCR and Real time PCR were studied. Result shows that gene expression does not exist in immature oocyte of sheep. In Tali goat, expression of this gene was more than in Rayeni goat.
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 goat oocytes and embryos. Immature oocytes (N = 90) were obtained from a local abattoir; mature oocytes (N = 40) and embryos (N = 25) were obtained by laparotomy from adult hair ewes submitted to ovarian stimulation treatment. The CD44 mRNA was detected by 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 Growth phase 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, in material from immature oocytes and cumulus cells. The CD44 amplicons were detected after a second heminested 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 goats.
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