Raman-microscopy investigation of vitrification-induced structural damages in mature bovine oocytes

PloS One
Giulia RuscianoBianca Gasparrini

Abstract

Although oocyte cryopreservation has great potentials in the field of reproductive technologies, it still is an open challenge in the majority of domestic animals and little is known on the biochemical transformation induced by this process in the different cellular compartments. Raman micro-spectroscopy allows the non-invasive evaluation of the molecular composition of cells, based on the inelastic scattering of laser photons by vibrating molecules. The aim of this work was to assess the biochemical modifications of both the zona pellucida and cytoplasm of vitrified/warmed in vitro matured bovine oocytes at different post-warming times. By taking advantage of Principal Component Analysis, we were able to shed light on the biochemical transformation induced by the cryogenic treatment, also pointing out the specific role of cryoprotective agents (CPs). Our results suggest that vitrification induces a transformation of the protein secondary structure from the α-helices to the β-sheet form, while lipids tend to assume a more packed configuration in the zona pellucida. Both modifications result in a mechanical hardening of this cellular compartment, which could account for the reduced fertility rates of vitrified oocytes. Furthermo...Continue Reading

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Citations

Jun 26, 2018·Reproduction in Domestic Animals = Zuchthygiene·Batool SanaeiMohammad H Nasr-Esfahani
Jun 24, 2019·European Biophysics Journal : EBJ·Elena GioloLaura Andolfi
Feb 24, 2020·Theriogenology·Luisa BoglioloSergio Ledda

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Methods Mentioned

BETA
FCS
PCA
Fluorescence

Software Mentioned

WiTech Project
Cryotop
princomp
MATLAB

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