HEPARAN SULFATE EXPRESSION IN MURINE OOCYTES

FARRANDO B; JULIANELLI VL; CALVO L; CALVO JC; ROMANATO M

Buenos Aires

Congreso; XII Jornadas Anuales de la Sociedad Argentina de Biología.; 2010

Sociedad Argentina de Biología

Previous studies from our laboratory demonstrated the role of Heparan sulfate (HS) as decondensing agent of human spermatozoa in vitro and its presence in mature mouse oocytes. The aim of this study was to determine at what time in follicular development the oocyte begins to express HS. Eight weeks old female CF1 mice were stimulated with PMSG and hCG (7.5 UI for both) to recover mature oocytes (MII) or with PMSG alone to retrieve immature oocytes. MII were recovered from the oviduct and cumulus cells were removed. Immature oocytes were recovered puncturing the ovary. In both cases, zona pellucida was removed and oocytes were fixed and permeabilized with formaldehyde and Tween-20. Immunocytochemistry was performed using anti-HS (monoclonal) as first antibody, anti-mouse IgM as second and propidium iodide to label DNA. Fluorescent label was observed both in MII and immature oocytes. Results demonstrate that HS is already present at early stages of follicular development.and its presence in mature mouse oocytes. The aim of this study was to determine at what time in follicular development the oocyte begins to express HS. Eight weeks old female CF1 mice were stimulated with PMSG and hCG (7.5 UI for both) to recover mature oocytes (MII) or with PMSG alone to retrieve immature oocytes. MII were recovered from the oviduct and cumulus cells were removed. Immature oocytes were recovered puncturing the ovary. In both cases, zona pellucida was removed and oocytes were fixed and permeabilized with formaldehyde and Tween-20. Immunocytochemistry was performed using anti-HS (monoclonal) as first antibody, anti-mouse IgM as second and propidium iodide to label DNA. Fluorescent label was observed both in MII and immature oocytes. Results demonstrate that HS is already present at early stages of follicular development.II) or with PMSG alone to retrieve immature oocytes. MII were recovered from the oviduct and cumulus cells were removed. Immature oocytes were recovered puncturing the ovary. In both cases, zona pellucida was removed and oocytes were fixed and permeabilized with formaldehyde and Tween-20. Immunocytochemistry was performed using anti-HS (monoclonal) as first antibody, anti-mouse IgM as second and propidium iodide to label DNA. Fluorescent label was observed both in MII and immature oocytes. Results demonstrate that HS is already present at early stages of follicular development.II were recovered from the oviduct and cumulus cells were removed. Immature oocytes were recovered puncturing the ovary. In both cases, zona pellucida was removed and oocytes were fixed and permeabilized with formaldehyde and Tween-20. Immunocytochemistry was performed using anti-HS (monoclonal) as first antibody, anti-mouse IgM as second and propidium iodide to label DNA. Fluorescent label was observed both in MII and immature oocytes. Results demonstrate that HS is already present at early stages of follicular development.