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Ultrastructure and molecular cytogenetics of human metaphase II oocytes with granular vacuoles

Published online by Cambridge University Press:  21 August 2009

Célia Soares ,
Ana Raquel Azevedo ,
Carolina Almeida ,
Elsa Oliveira ,
Ângela Alves ,
Joaquina Silva ,
Rosália Sá ,
Alberto Barros  and
Mário Sousa
Célia Soares
Affiliation:
Laboratory of Cell Biology, Institute of Biomedical Sciences Abel Salazar ofUniversity of Porto, Lg Prof Abel Salazar 2, 4009-003 Porto, Portugal
Ana Raquel Azevedo
Affiliation:
Department of Genetics, Faculty of Medicine ofUniversity of Porto, Alameda Prof. Hêrnani Monteiro, 4200-319 Porto, Portugal
Carolina Almeida
Affiliation:
Department of Genetics, Faculty of Medicine ofUniversity of Porto, Alameda Prof. Hêrnani Monteiro, 4200-319 Porto, Portugal
Elsa Oliveira
Affiliation:
Laboratory of Cell Biology, Institute of Biomedical Sciences Abel Salazar ofUniversity of Porto, Lg Prof Abel Salazar 2, 4009-003 Porto, Portugal
Ângela Alves
Affiliation:
Laboratory of Cell Biology, Institute of Biomedical Sciences Abel Salazar ofUniversity of Porto, Lg Prof Abel Salazar 2, 4009-003 Porto, Portugal
Joaquina Silva
Affiliation:
Centre for Reproductive Genetics Alberto Barros, Av. do Bessa 591 1º Dto. Frente, 4100-009 Porto, Portugal
Rosália Sá
Affiliation:
Laboratory of Cell Biology, Institute of Biomedical Sciences Abel Salazar ofUniversity of Porto, Lg Prof Abel Salazar 2, 4009-003 Porto, Portugal
Alberto Barros
Affiliation:
Centre for Reproductive Genetics Alberto Barros, Av. do Bessa 591 1º Dto. Frente, 4100-009 Porto, Portugal Department of Genetics, Faculty of Medicine ofUniversity of Porto, Alameda Prof. Hêrnani Monteiro, 4200-319 Porto, Portugal
Mário Sousa
Affiliation:
Laboratory of Cell Biology, Institute of Biomedical Sciences Abel Salazar ofUniversity of Porto, Lg Prof Abel Salazar 2, 4009-003 Porto, Portugal Centre for Reproductive Genetics Alberto Barros, Av. do Bessa 591 1º Dto. Frente, 4100-009 Porto, Portugal Department of Genetics, Faculty of Medicine ofUniversity of Porto, Alameda Prof. Hêrnani Monteiro, 4200-319 Porto, Portugal

Abstract

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Studies demonstrated poor fertilization rates and compromised early embryo development in cases of oocytes with dark and granular ooplasm, or with central granular ooplasm and presence of a refractile body. Many other structural abnormalities of human mature oocytes do exist, but remain to be evaluated. In the present study we characterized granular vacuoles (GVa) of mature metaphase II human oocytes. Surplus abnormal MII oocytes (n=6) with GVa were used under informed consent, after controlled superovulation during IVF treatment cycles. In all cases, patients had normal karyotypes. For electron microscopy oocytes were fixed with karnovsky fixative for 2h at 4°C, post-fixed in 1-2% OsO4 in buffer containing 0,8% K3Fe3+(CN)6, dehydrated in ethanol series, equilibrated with propylene oxide and embedded in Epon. Semithin sections were stained with aqueous azur II and methylene blue (1:1). Ultrathin sections were stained with 3% aquous uranyl acetate (20 min) and Reynols lead citrate (10 min). Sections were observed with a transmission electron microscope JEOL 100CXII operated at 60kV.

Type
Life Sciences
Information
Microscopy and Microanalysis , Volume 15 , Supplement S3 , July 2009 , pp. 31 - 32
Copyright
Copyright © Microscopy Society of America 2009