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Raman microspectroscopy as a non-invasive tool to assess the vitrification-induced changes of ovine oocyte zona pellucida

Bogliolo, Luisa and Ledda, Sergio and Innocenzi, Plinio and Ariu, Federica and Bebbere, Daniela and Rosati, Irma and Leoni, Giovanni Giuseppe and Piccinini, Massimo (2012) Raman microspectroscopy as a non-invasive tool to assess the vitrification-induced changes of ovine oocyte zona pellucida. Cryobiology, Vol. 64 (3), p. 267-272. eISSN 1090-2392. Article.

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DOI: 10.1016/j.cryobiol.2012.02.010

Abstract

Cryopreservation-induced modifications of zona pellucida (ZP) have been explored to a lesser extent compared to other oocyte compartments. Different methods have been applied to identify ZP changes, but most of them are invasive and measure only few properties of ZP.
Raman microspectroscopy (RMS) is a powerful technique for studying the molecular composition of cells but to date few studies have been performed on the oocytes using this method. The aim of the present study is to investigate the structural modifications of ZP of vitrified/warmed in vitro matured ovine oocytes by means of RMS. Cumulus–oocyte complexes were recovered from the ovaries of slaughtered adult sheep, matured in vitro and vitrified following the Minimum Essential Volume method using cryotops. ZPs of vitrified/warmed oocytes (VITRI), were exposed to vitrification solutions but not cryopreserved (CPA-exp) and untreated oocytes (CTR) were analyzed by RMS. We focused our analysis on the ZP protein and carbohydrate components by analyzing the 1230–1300 cm−1 amide III region and the 1020–1140 cm−1 spectral range in RMS spectra, respectively.
The spectral profiles in the ranges of proteins and carbohydrates were comparable between CTR and CPA-exp ZPs, whereas VITRI ZPs showed a significantly altered protein secondary structure characterized by an increase in β-sheet content and a decrease in the α-helix content. A significant modification of the carbohydrate components was also observed. This study demonstrates that vitrification of ovine oocytes induces biochemical changes of ZP related to the secondary structure of proteins and carbohydrate residues. Cryoprotectants do not strongly alter the molecular composition of ZP which is affected mainly by cooling. Raman technology offers a powerful and non-invasive tool to assess molecular modifications induced by cryopreservation in oocytes.

Item Type:Article
ID Code:8642
Status:Published
Refereed:Yes
Uncontrolled Keywords:Raman microspectroscopy, oocyte zona pellucida, vitrification
Subjects:Area 07 - Scienze agrarie e veterinarie > VET/10 Clinica ostetrica e ginecologia veterinaria
Area 05 - Scienze biologiche > BIO/10 Biochimica
Area 09 - Ingegneria industriale e dell'informazione > ING-IND/22 Scienza e tecnologia dei materiali
Divisions:002 Altri enti e centri di ricerca del Nord Sardegna > Porto Conte Ricerche, Alghero
001 Università di Sassari > 01-a Nuovi Dipartimenti dal 2012 > Architettura, Design e Urbanistica
001 Università di Sassari > 01-a Nuovi Dipartimenti dal 2012 > Medicina Veterinaria
Publisher:Academic Press / Elsevier
eISSN:1090-2392
Deposited On:19 Feb 2013 14:30

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