Transmission electron microscopy studies of the zona reaction in pig oocytes fertilized in vivo and in vitro

Hiroaki Funahashi, H. Ekwall, K. Kikuchi, H. Rodriguez-Martinez

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The aim of this study was to determine the ultrastructure of cross-sectioned zonae pellucidae of in vitro-matured and ovulated pig oocytes before or after sperm penetration in vitro and in vivo, respectively. The in vitro and in vivo (ovulated) oocytes and zygotes (fertilized in vitro and in vivo) were fixed with glutaraldehyde either directly or after pretreatment with ruthenium red and saponin, processed and then examined using transmission electron microscopy. The thickness of the zona pellucida, as measured on the section of the specimens with largest diameter fixed with glutaraldehyde, differed between the in vivo (9.19 ± 0.47 μm) and in vitro (5.95 ± 0.51 μm) oocytes. The in vivo oocytes had a rather thick external mesh-like structure, whereas it was much thinner in the in vitro oocytes. This mesh-like external rim was less apparent in both in vivo and in vitro zygotes. Obvious differences in the density of the lattice formed by the fixed zonae pellucidae were visible between the outer and inner (ad-oolemmal) zonae. The outer area always formed a concentrically arrayed fibrillar network, whereas the inner area showed a much more compact, trabecule-like mesh. However, both areas, but particularly the outer network, were much more compacted after the zona reaction. Clear differences in the degree of fibrillar aggregation of the inner zona area were also observed between in vitro and in vivo zygotes, being much higher in the latter. This fibrillar network was more clearly visible in the zygotes pretreated with ruthenium red and saponin; the in vitro zygotes had a fibrillar, radially oriented set of parallel fibrils, whereas it was much more aggregated and trabecule-like in the in vivo zygotes. These results demonstrate that the fine structure of the zona pellucida and the zona reaction at sperm penetration differ between pig oocytes fertilized in vivo and in vitro. Moreover, the ultrastructure of the outer and inner pig zonae pellucidae has a different network organization.

Original languageEnglish
Pages (from-to)443-452
Number of pages10
JournalReproduction
Volume122
Issue number3
Publication statusPublished - 2001
Externally publishedYes

Fingerprint

Herpes Zoster
Transmission Electron Microscopy
Oocytes
Swine
Zygote
Zona Pellucida
Sperm-Ovum Interactions
Ruthenium Red
Saponins
Glutaral
In Vitro Techniques

ASJC Scopus subject areas

  • Obstetrics and Gynaecology
  • Cell Biology
  • Endocrinology
  • Embryology

Cite this

Transmission electron microscopy studies of the zona reaction in pig oocytes fertilized in vivo and in vitro. / Funahashi, Hiroaki; Ekwall, H.; Kikuchi, K.; Rodriguez-Martinez, H.

In: Reproduction, Vol. 122, No. 3, 2001, p. 443-452.

Research output: Contribution to journalArticle

Funahashi, H, Ekwall, H, Kikuchi, K & Rodriguez-Martinez, H 2001, 'Transmission electron microscopy studies of the zona reaction in pig oocytes fertilized in vivo and in vitro', Reproduction, vol. 122, no. 3, pp. 443-452.
Funahashi, Hiroaki ; Ekwall, H. ; Kikuchi, K. ; Rodriguez-Martinez, H. / Transmission electron microscopy studies of the zona reaction in pig oocytes fertilized in vivo and in vitro. In: Reproduction. 2001 ; Vol. 122, No. 3. pp. 443-452.
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