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The hyaluronic acid receptor (CD44) is expressed in bovine oocytes and early stage embryos

Furnus, Cecilia C., Valcarcel, Alberto, Dulout, Fernando N., Errecalde, Ana Lía
Theriogenology 2003 v.60 no.9 pp. 1633-1644
animal tissues, blastocyst, cattle, cell membranes, cytoplasm, developmental stages, eggs, embryogenesis, embryonic stem cells, extracellular matrix, fluids, fluorescein, fluorescent antibody technique, humans, hyaluronic acid, in vitro fertilization, isomers, mice, molecular weight, oocytes, spermatozoa, swine, umbilical cord
Hyaluronic acid (HA) is a high molecular weight polysaccharide found in the extracellular matrix of most animal tissues, that exerts a profound influence on cell behavior. HA is one of the most abundant glycosaminoglycans (GAGs) in the uterine, oviductal and follicular fluids in mouse, pig, human and cattle. CD44, the principal cell membrane receptor for HA, is expressed from the 1- to 8-cell stage in human embryos, during post-implantation mouse embryogenesis and on the surface of differentiated embryonic stem cells. In the present study, we have analyzed by immunofluorescence, whether CD44 is present in bovine oocytes, fertilized oocytes and early stage embryos. Bovine cumulus-oocyte complexes (COCs) were aspirated from follicles (2-5mm) and were selected for IVM and incubated for 24h. Oocytes showing an expanded cumulus (generally 90-95%) were used for IVF. Fertilized oocytes were separated for immunofluorescence assay after 16h of sperm incubation in order to fix the eggs at the pronuclear stage. The embryos were cultured for 8 days and the different stages of development for immunofluorescence assay were separated every 24h of culture. The CD44 receptor was detected at every observation time examined. Fluorescence-tagged HA for the internalization assay was prepared by mixing fluorescein amine, Isomer I and 1mg of HA from umbilical cord. Fluorescence-tagged HA was internalized in 2-, 4-, 8- and 16-cell-stage embryos, morulae and blastocysts. CD44 is expressed on the surface and in the cytoplasm of bovine oocytes and embryos in different stages of development.