Greener Journal of Biological Sciences Vol. 4 (5), pp. 139-145, June 2014.
© 2014 Authors
Manuscript Number: 050614223
Expression of CD44 Gene in Goat’s Oocytes and Embryos
M. Aminafshar1, V. Bahrampour1*, A. Bagizadeh2,
N. Emam Jomeh Kashan1 and M.R. Mohamad Abadi3
1Department of Animal Science and Science and Research Branch, Islamic Azad University, Tehran, Iran.
2Department of Gentic and Advanced Sciences University, Kerman, Iran.
3Department of animal Science and Shahid Bahonar University, Kerman, Iran.
*Corresponding Author’s Email: v_bahrampur @yahoo .com
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 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 goats.
Keywords: Gene expression, Hemi nested PCR, Goat, Oocyte, Embryos.
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