Confocal and fluorescence microscopic study using lectins of the distribution of cortical granules during the maturation and fertilization of pig oocytes.
A study was carried out to determine whether pig cortical granules (CGs) could be visualized using fluorescein isothiocyanate (FITC)-labelled lectins. Following labelling with FITC-labelled peanut agglutinin (FITC-PNA), fluorescent spots were observed that had a distribution during maturation and fertilization entirely consistent with that observed by electron microscopy. For the first 18 h of in vitro maturation, most of the fluorescent spots of FITC-PNA were distributed throughout the cortical cytoplasm. Thereafter, the CGs underwent centrifugal migration to form a monolayer next to the plasma membrane. Following penetration by sperm, fluorescent spots were extruded into the perivitelline space, where they aggregated forming fluorescent clumps, which subsequently formed a reticulate structure surrounding the egg. Fluorescence was gradually lost such that by 18 h after insemination none could be detected in 70% of the eggs. The results indicate that CGs in pig oocytes contain galactosyl-rich glycoconjugates and that FITC-PNA is a useful probe for their rapid visualization and examination.
Yoshida
M
M
Department of Molecular Embryology, AFRC Institute of Animal Physiology and Genetics Research, Cambridge, United Kingdom.
Cran
D G
DG
Pursel
V G
VG
D000818:Animals; D002240:Carbohydrate Sequence; D003594:Cytoplasmic Granules / Q000378:metabolism / Q000648:ultrastructure; D004187:Disaccharides / Q000737:chemistry / Q000378:metabolism; D005260:Female; D005306:Fertilization; D005307:Fertilization in Vitro; D016650:Fluorescein-5-isothiocyanate; D006001:Glycoconjugates / Q000378:metabolism; D006651:Histocytochemistry; D066298:In Vitro Techniques; D037102:Lectins / Q000378:metabolism; D008297:Male; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009865:Oocytes / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure*; D019887:Peanut Agglutinin; D011247:Pregnancy; D013552:Swine
D004187:Disaccharides; D006001:Glycoconjugates; D037102:Lectins; D019887:Peanut Agglutinin; D016650:Fluorescein-5-isothiocyanate