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Raman-microscopy investigation of vitrification-induced structural damages in mature bovine oocytes
- Source :
- PLoS ONE, Vol 12, Iss 5, p e0177677 (2017), PLoS ONE
- Publication Year :
- 2017
- Publisher :
- Public Library of Science (PLoS), 2017.
-
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. Furthermore, biochemical modifications were observed at the cytoplasmic level in the protein secondary structure, with α-helices loss, suggesting cold protein denaturation. In addition, a decrease of lipid unsaturation was found in vitrified oocytes, suggesting oxidative damages. Interestingly, most modifications were not observed in oocytes exposed to CPs, suggesting that they do not severely affect the biochemical architecture of the oocyte. Nevertheless, in oocytes exposed to CPs decreased developmental competence and increased reactive oxygen species production were observed compared to the control. A more severe reduction of cleavage and blastocyst rates after in vitro fertilization was obtained from vitrified oocytes. Our experimental outcomes also suggest a certain degree of reversibility of the induced transformations, which renders vitrified oocytes more similar to untreated cells after 2 h warming.
- Subjects :
- 0301 basic medicine
Cytoplasm
Embryology
lcsh:Medicine
Reproductive technology
Spectrum Analysis, Raman
Biochemistry
Oogenesis
Protein Structure, Secondary
Cryopreservation
Oxidative Damage
0302 clinical medicine
Animal Cells
Macromolecular Structure Analysis
Vitrification
Birth Rate
Zona pellucida
lcsh:Science
Principal Component Analysis
030219 obstetrics & reproductive medicine
Multidisciplinary
Fertility Preservation
Lipids
Cell biology
medicine.anatomical_structure
OVA
bovine oocytes, vitrification, Raman-microscopy
Female
Structural Proteins
Cellular Types
Cellular Structures and Organelles
Research Article
Protein Structure
Cell Survival
Specimen Preservation
Biology
Research and Analysis Methods
Cryobiology
03 medical and health sciences
Botany
medicine
Animals
Blastocyst
Molecular Biology
Zona Pellucida
lcsh:R
Biology and Life Sciences
Proteins
Cell Biology
Oocyte cryopreservation
Lipid Metabolism
Oocyte
Germ Cells
030104 developmental biology
Specimen Preparation and Treatment
Oocytes
Blastocysts
Cattle
lcsh:Q
Reactive Oxygen Species
Developmental Biology
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 12
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....d8c460c5e830c968ab20aa98ddcbf701