Greener Journal of Biological Sciences

Open Access

Subscribe to 

our monthly News letters

Greener Journal of  Biological Sciences Vol. 4 (5), pp. 139-145, June 2014.

 © 2011 Greener Journals

Research Paper

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.


Allworth AE; Albertini DF (1993). Meiotic maturation in cultured bovine oocytes is accompanied by remodeling of the cumulus cell cytoskeleton. Developmental biology. 158: 101-112.

Archibong AE, Petters RM; Johnson BH (1989). Development of porcine embryos from one- and two-cell stages to blastocysts in culture medium supplemented with porcine oviductal fluid. Biology of reproduction. 41: 1076-1083.

Assidi M, Dufort I, Ali A, Hamel M (2008). Identification of potential markers of oocyte competence expressed in bovine cumulus cells matured with follicle-stimulating hormone and/or phorbol myristate acetate in vitro. Biology of reproduction. 79: 209-222.

Borg N ; Holland M (2008). The effect of glycosaminoglycans on rat gametes in vitro and the associated signal pathway. Reproduction 135: 311-319.

Campbell S, Swann HR, Aplin JD, Seif MW, et al. (1995). CD44 is expressed throughout pre-implantation human embryo development. Human Reproduction. 10: 425-430.

Chen L, Russell PT; Larsen WJ (1993). Functional significance of cumulus expansion in the mouse: roles for the preovulatory synthesis of hyaluronic acid within the cumulus mass. Molecular reproduction. 34: 87-93.

Daum G, Eisenmann-Tappe I, Fries HW, Troppmair J (1994). The ins and outs of Raf kinases. Trends in biochemical. 19: 474-480.

Fulop C, Salustri A ; Hascall VC (1997). Coding sequence of a hyaluronan synthase homologue expressed during expansion of the mouse cumulus-oocyte complex. Human reproduction. 337: 261-266.

Furnus CC, de Matos DG ; Martinez AG (1998). Effect of hyaluronic acid on development of in vitro produced bovine embryos. Theriogenology. 49: 1489-1499.

Furnus CC, Valcarcel A, Dulout FN ; Errecalde AL (2003). The hyaluronic acid receptor (CD44) is expressed in bovine oocytes and early stage embryos. Theriogenology. 60: 1633-1644.

Goodison S, Urquidi V ; Tarin D (1999). CD44 cell adhesion molecules. Molecular Pathology. 52: 189-196.

Inoue M, Naito K, Nakayama T and Sato E (1998). Mitogen-activated protein kinase translocates into the germinal vesicle and induces germinal vesicle breakdown in porcine oocytes. Biology of reproduction. 58: 130-136.

Jackson RL, Busch SJ ; Cardin AD (1991). Glycosaminoglycans: molecular properties, protein interactions, and role in physiological processes. cellular physiology. 71: 481-539.

Kaya G, Laurini R, Chaubert P ; Gross N (2001). Expression of CD44 and its isoforms in the fetal neuroblasts.. Morpholgy. 9: 180-184.

Kimura N, Konno Y, Miyoshi K, Matsumoto H, et al. (2002). Expression of hyaluronan synthases and CD44 messenger RNAs in porcine cumulus-oocyte complexes during in vitro maturation. Biology Reported. 66: 707-717.

Kimura N, Hoshino Y, Totsukawa K ; Sato E (2007). Cellular and molecular events during oocyte maturation in mammals: molecules of cumulus-oocyte complex matrix and signalling pathways regulating meiotic progression. Society of Reproduction. 63: 327-342.

Lee CN ; Ax RL (1984). Concentrations and composition of glycosaminoglycans in the female bovine reproductive tract. J. Dairy Sci. 67: 2006-2009.

Li HK, Kuo TY, Yang HS, Chen LR, et al. (2008). Differential gene expression of bone morphogenetic protein 15 and growth differentiation factor 9 during in vitro maturation of porcine oocytes and early embryos. Animal reproduction. 103: 312-322.

Miyano T, Hiro-Oka RE, Kano K, Miyake M, (1994). Effects of hyaluronic acid on the development of 1- and 2-cell porcine embryos to the blastocyst stage in vitro. Theriogenology. 41: 1299-1305.

Ohta N, Saito H, Kaneko T, Yoshida M (2001). Soluble CD44 in human ovarian follicular fluid. J Assist. assisted reproduction. 18: 21-25.

Peterson PE, Pow CS, Wilson DB ; Hendrickx AG (1993). Distribution of extracellular matrix components during early embryonic development in the macaque. Cells Tissues. 146: 3-13.

Salustri A, Yanagishita M ; Hascall VC (1989). Synthesis and accumulation of hyaluronic acid and proteoglycans in the mouse cumulus cell-oocyte complex during follicle-stimulating hormone-induced mucification. biology Chemictry. 264: 13840-13847.

Salustri A, Ulisse S, Yanagishita M ; Hascall VC (1990). Hyaluronic acid synthesis by mural granulosa cells and cumulus cells in vitro is selectively stimulated by a factor produced by oocytes and by transforming growth factor-b. biology Chemictry. 265: 19517-19523.

Sato E, Ishibashi T and Koide SS (1987). Prevention of spontaneous degeneration of mouse oocytes in culture by ovarian glycosaminoglycans. Biology Reported. 37: 371-376.

Sato E, Inoue M, Takahashi Y ; Toyoda Y (1994). Glycosaminoglycans prevent induction of fragmentation of porcine oocytes stimulated by dibutyryl cyclic adenosine 3’,5’-monophosphate in culture. Cell structure and function. 19: 29-36.

Schoenfelder M ; Einspanier R (2003). Expression of hyaluronan synthases and corresponding hyaluronan receptors is differentially regulated during oocyte maturation in cattle. Biolog Reported. 69: 269-277.

Slevin M, Krupinski J, Kumar S ; Gaffney J (1998). Angiogenic oligosaccharides of hyaluronan induce protein tyrosine kinase activity in endothelial cells and activate a cytoplasmic signal transduction pathway resulting in proliferation. Laboratory investigation. 78: 987-1003.

Toyokawa K, Harayama H ; Miyake M (2005). Exogenous hyaluronic acid enhances porcine parthenogenetic embryo development in vitro possibly mediated by CD44. Theriogenology. 64: 378-392.

Tunjung WA, Yokoo M, Hoshino Y, Miyake Y (2009). Effect of hyaluronan to inhibit caspase activation in porcine granulosa cells. Biochemical and Biophysical Research Communications. 382: 160-164.

Wheatley SC, Isacke CM ; Crossley PH (1993). Restricted expression of the hyaluronan receptor, CD44, during postimplantation mouse embryogenesis suggests key roles in tissue formation and patterning. Development. 119: 295-306.

Yokoo M, Miyahayashi Y, Naganuma T, Kimura N, et al. (2002). Identification of hyaluronic acid-binding proteins and their expressions in porcine cumulus-oocyte complexes during in vitro maturation. Biology Reported. 67: 1165-1171.

Yokoo M, Shimizu T, Kimura N, Tunjung WA, et al. (2007). Role of the hyaluronan receptor CD44 during porcine oocyte maturation. Reproduction in Domestic Animals. 53: 263-270.

Yokoo M, Kimura N ; Sato E (2010). Induction of oocyte maturation by hyaluronan-CD44 interaction in pigs. J. Reproduction in Domestic Animals. Dev. 56: 15-19.