Distribution of prepubertal and adult goat oocyte cortical granules during meiotic maturation and fertilisation: Ultrastructural and cytochemical study

Molecular Reproduction and Development - Tập 68 Số 4 - Trang 507-514 - 2004
E. Velilla1, D. Izquierdo1, E. Rodríguez‐González1, Manel López‐Béjar2, Francesca Vidal3, M.T. Paramio4
1Departament de Ciència Animal i dels Aliments, Facultat de Vetèrinaria, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain
2Departament de Sanitat i Anatomia Animals, Facultat de Vetèrinaria, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain
3Departament de Biologia Cellular, Fisiologia i Immunologia, Facultat de Ciències, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain
4Departament de Ciència Animal i dels Aliments, Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain

Tóm tắt

AbstractThe aim of this study was evaluate cortical granule (CG) distribution during in vitro maturation (IVM) and fertilisation of prepubertal goat oocytes compared to CG distribution of ovulated and in vitro fertilised oocytes from adult goats. Oocytes from prepubertal goats were recovered from a slaughterhouse and were matured in M199 with hormones and serum for 27 hr. Ovulated oocytes were collected from gonadotrophin treated Murciana goats. Frozen‐thawed spermatozoa were selected by centrifugation in percoll gradient and were capacitated in DMH with 20% steer serum for 1 hr. Ovulated and IVM‐oocytes were inseminated in DMH medium with steer serum and calcium lactate for 20 hr. Oocytes and presumptive zygotes were stained with FITC‐LCA (Lens culinaris agglutinin labelled with fluorescein isothiocyanate) and observed under a confocal laser scanning microscope. Ultrastructure morphology of oocytes and presumptive zygotes were analysed by transmission electron microscopy (TEM). Prepubertal goat oocytes at germinal vesicle stage show a homogeneous CG distribution in the cytoplasm. IVM‐oocytes at Metaphase II (MII) and ovulated oocytes presented CGs located in the cortex with the formation of a monolayer beneath to the plasma membrane. At 20 hr postinsemination (hpi), zygotes from IVM‐oocytes showed a complete CG exocytosis whereas zygotes from ovulated oocytes presented aggregates of CGs located at the cortical region. Images by TEM detected that CGs were more electrodense and compacts in oocytes from prepubertal than from adult goats. Mol. Reprod. Dev. 68: 507–514, 2004. © 2004 Wiley‐Liss, Inc.

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Molecular Reproduction and Development

Tập 68 Số 4

507-514

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