Whole-depth change in bovine zona pellucida biomechanics after fertilization: how relevant in hindering polyspermy?

Polyspermy is a common problem in bovine in vitro fertilization (IVF) and has a still unclear etiology. In this specie, after IVF, despite the lack of a biochemical post-fertilization hardening, the stiffness of the outer ZP layer is significantly increased. Therefore, polyspermy might be related to...

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Published in:PloS one 2012-09, Vol.7 (9), p.e45696-e45696
Main Authors: Papi, Massimiliano, Brunelli, Roberto, Familiari, Giuseppe, Frassanito, Maria Cristina, Lamberti, Luciano, Maulucci, Giuseppe, Monaci, Maurizio, Pappalettere, Carmine, Parasassi, Tiziana, Relucenti, Michela, Sylla, Lakamy, Ursini, Fulvio, De Spirito, Marco
Format: Article
Language:English
Subjects:
Animals
Atomic beam spectroscopy
Biology
Biomechanical Phenomena
Biomechanics
Cattle
Cytoplasm - metabolism
Electron microscopy
Etiology
Female
Fertility
Fertilization
Fertilization in Vitro
Filaments
Flagella
Flagella - metabolism
Glycoproteins - chemistry
In vitro fertilization
Male
Mammals
Maturation
Medicine
Microscopy
Microscopy, Atomic Force - methods
Microscopy, Electron, Transmission - methods
Models, Statistical
Oocytes - cytology
Oocytes - ultrastructure
Physiological aspects
Proteins
Ruthenium
Ruthenium red
Ruthenium Red - pharmacology
Spectroscopy
Spectrum analysis
Sperm
Spermatozoa - cytology
Spermatozoa - pathology
Stiffening
Stiffness
Transmission electron microscopy
Zona pellucida
Zona Pellucida - metabolism
Zona Pellucida - ultrastructure
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container_end_page e45696
container_issue 9
container_start_page e45696
container_title PloS one
container_volume 7
creator Papi, Massimiliano
Brunelli, Roberto
Familiari, Giuseppe
Frassanito, Maria Cristina
Lamberti, Luciano
Maulucci, Giuseppe
Monaci, Maurizio
Pappalettere, Carmine
Parasassi, Tiziana
Relucenti, Michela
Sylla, Lakamy
Ursini, Fulvio
De Spirito, Marco
description Polyspermy is a common problem in bovine in vitro fertilization (IVF) and has a still unclear etiology. In this specie, after IVF, despite the lack of a biochemical post-fertilization hardening, the stiffness of the outer ZP layer is significantly increased. Therefore, polyspermy might be related to an incomplete or insufficient stiffening of the ZP. We obtained, by using atomic force spectroscopy in physiological conditions, a complete characterization of the biomechanical changes of the inner and outer ZP layers occurring during oocyte maturation/fertilization and correlated them to the ultrastructural changes observed by transmission electron microscopy using ruthenium red and saponin technique. In both the inner and outer ZP layers, stiffness decreased at maturation while, conversely, increased after fertilization. Contextually, at the nanoscale, during maturation both ZP layers displayed a fine filaments network whose length increased while thickness decreased. After fertilization, filaments partially recovered the immature features, appearing again shorter and thicker. Overall, the observed biomechanical modifications were substantiated by ultrastructural findings in the ZP filament mesh. In fertilized ZP, the calculated force necessary to displace ZP filaments resulted quite similar to that previously reported as generated by bovine sperm flagellum. Therefore, in bovine IVF biomechanical modifications of ZP appear ineffective in hindering sperm transit, highlighting the relevance of additional mechanisms operating in vivo.
doi_str_mv 10.1371/journal.pone.0045696
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1932-6203
language eng
recordid cdi_plos_journals_1326551966
source Open Access: PubMed Central; Publicly Available Content Database
subjects Animals
Atomic beam spectroscopy
Biology
Biomechanical Phenomena
Biomechanics
Cattle
Cytoplasm - metabolism
Electron microscopy
Etiology
Female
Fertility
Fertilization
Fertilization in Vitro
Filaments
Flagella
Flagella - metabolism
Glycoproteins - chemistry
In vitro fertilization
Male
Mammals
Maturation
Medicine
Microscopy
Microscopy, Atomic Force - methods
Microscopy, Electron, Transmission - methods
Models, Statistical
Oocytes - cytology
Oocytes - ultrastructure
Physiological aspects
Proteins
Ruthenium
Ruthenium red
Ruthenium Red - pharmacology
Spectroscopy
Spectrum analysis
Sperm
Spermatozoa - cytology
Spermatozoa - pathology
Stiffening
Stiffness
Transmission electron microscopy
Zona pellucida
Zona Pellucida - metabolism
Zona Pellucida - ultrastructure
title Whole-depth change in bovine zona pellucida biomechanics after fertilization: how relevant in hindering polyspermy?
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