Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Journal of Assisted Reproduction and Genetics
Published in: Journal of Assisted Reproduction and Genetics 2/2015

01-02-2015 | Gamete Biology

Evaluation of the impact of vitrification on the actin cytoskeleton of in vitro matured ovine oocytes by means of Raman microspectroscopy

Authors: Luisa Bogliolo, Ombretta Murrone, Massimo Piccinini, Federica Ariu, Sergio Ledda, Sara Tilocca, David F. Albertini

Published in: Journal of Assisted Reproduction and Genetics | Issue 2/2015

Login to get access

Abstract

Purpose

Investigation of the changes induced by vitrification on the cortical F-actin of in vitro matured ovine oocytes by Raman microspectroscopy (RMS).

Methods

Cumulus-oocyte complexes, recovered from the ovaries of slaughtered sheep, were matured in vitro and vitrified following the Minimum Essential Volume method using cryotops. The cortical region of metaphase II (MII) oocytes (1) exposed to vitrification solutions but not cryopreserved (CPA-exp), (2) vitrified/warmed (VITRI), and (3) untreated (CTR) was analyzed by RMS. A chemical map of one quadrant of single CPA-exp, VITRI and CTR oocytes was, also, performed. In order to identify the region of Raman spectra representative of the cortical F-actin modification, a group of in vitro matured oocytes were incubated with latrunculin–A (LATA), a specific F-actin destabilizing drug, and processed for RMS analysis. Thereafter, all the oocytes were stained with rhodamine phalloidin and evaluated by fluorescence confocal microscopy. Raman spectra of the oocytes were, statistically, analyzed using Principal Component Analysis (PCA).

Results

The PCA score plots showed a marked discrimination between CTR oocytes and CPA-exp/ VITRI groups. The main differences, highlighted by PCA loadings, were referable to proteins (1657, 1440 and 1300 cm−1) and, as indicated by LATA experiments, also included the changes of the F-actin. Analysis by confocal microscopy revealed a clear alteration of the cortical F-actin of CPA-exp and VITRI oocytes confirming RMS results.

Conclusions

Raman microspectroscopy may represent an alternative analytical tool for investigating the biochemical modification of the oocyte cortex, including the F-actin cytoskeleton, during vitrification of in vitro matured ovine oocytes.
Literature
1.
Konc J, Kanyó K, Kriston R, Somoskői B, Cseh S. Cryopreservation of embryos and oocytes in human assisted reproduction. Biomed Res Int. 2014;2014:307268.CrossRefPubMedCentralPubMed
2.
Saragusty J, Arav A. Current progress in oocyte and embryo cryopreservation by slow freezing and vitrification. Reproduction. 2011;141:1–19.CrossRefPubMed
3.
Mullen SF, Fahy GM. A chronologic review of mature oocyte vitrification research in cattle, pigs, and sheep. Theriogenology. 2012;78:1709–19.CrossRefPubMed
4.
The Practice Committees of the American Society for Reproductive Medicine and the Society for Assisted Reproductive Technology. Mature oocyte cryopreservation: a guideline. Fertil Steril. 2013;99:37–43.CrossRef
5.
Arav A, Natan Y. Vitrification of oocytes: from basic science to clinical application. Adv Exp Med Biol. 2013;761:69–83.CrossRefPubMed
6.
Díez C, Muñoz M, Caamaño JN, Gómez E. Cryopreservation of the bovine oocyte: current status and perspectives. Reprod Domest Anim. 2012;3:76–83.CrossRef
7.
Vincent C, Johnson MH. Cooling, cryoprotectants, and the cytoskeleton of the mammalian oocyte. Oxf Rev Reprod Biol. 1992;14:73–100.PubMed
8.
Moussa M, Shu J, Zhang X, Zeng F. Cryopreservation of mammalian oocytes and embryos: current problems and future perspectives. Sci China Life Sci. 2014;57:903–14.CrossRefPubMed
9.
Li R, Albertini DF. The road to maturation: somatic cell interaction and self-organization of the mammalian oocyte. Nat Rev Mol Cell Biol. 2013;14:141–52.CrossRefPubMed
10.
Simerly C, Navara CS, Wu GJ, Schatten G. Cytoskeletal organization and dynamics in mammalian oocytes during maturation and fertilization. In: Grudzinskas JG, Yovich JL, editors. Gametes - the oocyte. Cambridge: Cambridge University Press; 1995. p. 54–94.
11.
Lei T, Guo N, Liu JQ, Tan MH, Li YF. Vitrification of in vitro matured oocytes: effects on meiotic spindle configuration and mitochondrial function. Int J Clin Exp Pathol. 2014;7:1159–65.PubMedCentralPubMed
12.
Tamura AN, Huang TT, Marikawa Y. Impact of vitrification on the meiotic spindle and components of the microtubule-organizing center in mouse mature oocytes. Biol Reprod. 2013;89:112.CrossRefPubMedCentralPubMed
13.
Egerszegi I, Somfai T, Nakai M, Tanihara F, Noguchi J, Kaneko H, et al. Comparison of cytoskeletal integrity, fertilization and developmental competence of oocytes vitrified before or after in vitro maturation in a porcine model. Cryobiology. 2013;67:287–92.CrossRefPubMed
14.
Coticchio G, Bromfield JJ, Sciajno R, Gambardella A, Scaravelli G, Borini A, et al. Vitrification may increase the rate of chromosome misalignment in the metaphase II spindle of human mature oocytes. Reprod Biomed Online. 2009;19:29–34.CrossRefPubMed
15.
Chen SU, Yang YS. Slow freezing or vitrification of oocytes: their effects on survival and meiotic spindles, and the time schedule for clinical practice. Taiwan J Obstet Gynecol. 2009;48:15–22.CrossRefPubMed
16.
Succu S, Leoni GG, Bebbere D, Berlinguer F, Mossa F, Bogliolo L, et al. Vitrification devices affect structural and molecular status of in vitro matured ovine oocytes. Mol Reprod Dev. 2007;74:1337–44.CrossRefPubMed
17.
Mikołajewska N, Müller K, Niżański W, Jewgenow K. Vitrification of domestic cat oocytes–effect on viability and integrity of subcellular structures. Reprod Domest Anim. 2012;47:295–9.CrossRefPubMed
18.
Wu C, Rui R, Dai J, Zhang C, Ju S, Xie B, et al. Effects of cryopreservation on the developmental competence, ultrastructure and cytoskeletal structure of porcine oocytes. Mol Reprod Dev. 2006;73:1454–62.CrossRefPubMed
19.
Rojas C, Palomo MJ, Albarracín JL, Mogas T. Vitrification of immature and in vitro matured pig oocytes: study of distribution of chromosomes, microtubules, and actin microfilaments. Cryobiology. 2004;49:211–20.CrossRefPubMed
20.
Hotamisligil S, Toner M, Powers RD. Changes in membrane integrity, cytoskeletal structure, and developmental potential of murine oocytes after vitrification in ethylene glycol. Biol Reprod. 1996;55:161–8.CrossRefPubMed
21.
Combelles CM, Ceyhan ST, Wang H, Racowsky C. Maturation outcomes are improved following Cryoleaf vitrification of immature human oocytes when compared to choline-based slow-freezing. J Assist Reprod Genet. 2011;28:1183–92.CrossRefPubMedCentralPubMed
22.
Azoury J, Lee KW, Georget V, Hikal P, Verlhac MH. Symmetry breaking in mouse oocytes requires transient F-actin meshwork destabilization. Development. 2011;138:2903–8.CrossRefPubMed
23.
Khalili MA, Maione M, Palmerini MG, Bianchi S, Macchiarelli G, Nottola SA. Ultrastructure of human mature oocytes after vitrification. Eur J Histochem. 2012;56:e38.CrossRefPubMedCentralPubMed
24.
Caamaño JN, Muñoz M, Diez C, Gómez E. Polarized light microscopy in mammalian oocytes. Reprod Domest Anim. 2010;2:49–56.CrossRef
25.
Coticchio G, Sciajno R, Hutt K, Bromfield J, Borini A, Albertini DF. Comparative analysis of the metaphase II spindle of human oocytes through polarized light and high-performance confocal microscopy. Fertil Steril. 2010;93:2056–64.CrossRefPubMed
26.
Notingher I, Hench LL. Raman microspectroscopy: a noninvasive tool for studies of individual living cells in vitro. Expert Rev Med Devices. 2006;3:215–34.CrossRefPubMed
27.
28.
Wood BR, Chernenko T, Matthaus C, Diem M, Chong C, Bernhard U, et al. Shedding new light on the molecular architecture of oocytes using a combination of synchrotron fourier transm-infrared and Raman spectroscopic mapping. Anal Chem. 2008;80:9065–72.CrossRefPubMedCentralPubMed
29.
Davidson B, Murray AA, Elfick A, Spears N. Raman micro-spectroscopy can be used to investigate the developmental stage of the mouse oocyte. PLoS One. 2013;8:e67972.CrossRefPubMedCentralPubMed
30.
Davidson B, Spears N, Murray A, Elfick A. The changing biochemical composition and organisation of the murine oocyte and early embryo as revealed by Raman spectroscopic mapping. J Raman Spectrosc. 2012;43:24–31.CrossRef
31.
Bogliolo L, Ledda S, Innocenzi P, Ariu F, Bebbere D, Rosati I, et al. Raman microspectroscopy as a non-invasive tool to assess the vitrification-induced changes of ovine oocyte zona pellucida. Cryobiology. 2012;64:267–72.CrossRefPubMed
32.
Bogliolo L, Murrone O, Di Emidio G, Piccinini M, Ariu F, Ledda S, et al. Raman spectroscopy-based approach to detect aging-related oxidative damage in the mouse oocyte. J Assist Reprod Genet. 2013;30:877–82.CrossRefPubMedCentralPubMed
33.
Spector I, Shochet NR, Blasberger D, Kashman Y. Latrunculins–novel marine macrolides that disrupt microfilament organization and affect cell growth: I. Comparison with cytochalasin D. Cell Motil Cytoskeleton. 1989;13:127–44.CrossRefPubMed
34.
Spector I, Shochet NR, Kashman Y, Groweiss A. Latrunculins: novel marine toxins that disrupt microfilament organization in cultured cells. Science. 1983;219:493–5.CrossRefPubMed
35.
Kuwayama M, Vajta G, Kato O, Leibo SP. Highly efficient vitrification method for cryopreservation of human oocytes. Reprod BioMed Online. 2005;11:300–8.CrossRefPubMed
36.
Zoladek AB, Johal RK, Garcia-Nieto S, Pascut F, Shakesheff KM, Ghaemmaghami AM. Notingher Label-free molecular imaging of immunological synapses between dendritic and T cells by Raman micro-spectroscopy. Analyst. 2010;135:3205–12.CrossRefPubMed
37.
Matthäus C, Chernenko T, Newmark JA, Warner CM, Diem M. Label-free detection of mitochondrial distribution in cells by nonresonant Raman microspectroscopy. Biophys J. 2007;93:668–73.CrossRefPubMedCentralPubMed
38.
Kim SS, Olsen R, Kim DD, Albertini DF. The impact of vitrification on immature oocyte cell cycle and cytoskeletal integrity in a rat model. J Assist Reprod Genet. 2014;31:739–47.CrossRefPubMed
39.
Albarracín JL, Morató R, Rojas C, Mogas T. Effects of vitrification in open pulled straws on the cytology of in vitro matured prepubertal and adult bovine oocytes. Theriogenology. 2005;63:890–901.CrossRefPubMed
40.
Lee K, Wang C, Spate L, Murphy CN, Prather RS, Machaty Z. Gynogenetic activation of porcine oocytes. Cell Reprogram. 2014;16:121–9.CrossRefPubMed
41.
Maro B, Johnson MH, Pickering SJ, Flach G. Changes in actin distribution during fertilization of the mouse egg. J Embryol Exp Morphol. 1984;81:211–37.PubMed
42.
Lee K, Wang C, Spate L, Murphy CN, Prather RS, Machaty Z. Gynogenetic activation of porcine oocytes. Cell Reprogram. 2014;16:121–9.CrossRefPubMed
43.
Hosu BG, Mullen SF, Critser JK, Forgacs G. Reversible disassembly of the actin cytoskeleton improves the survival rate and developmental competence of cryopreserved mouse oocytes. PLoS One. 2008;30:3–e2787.
44.
Wen Y, Zhao S, Chao L, Yu H, Song C, Shen Y, Chen H, Deng X. The protective role of antifreeze protein 3 on the structure and function of mature mouse oocytes in vitrification. Cryobiology. 2014, in press.
45.
46.
Riedl J, Crevenna AH, Kessenbrock K, Yu JH, Neukirchen D, Bista M, et al. Lifeact: a versatile marker to visualize F-actin. Nat Methods. 2008;5:605–7.CrossRefPubMedCentralPubMed
47.
Chaigne A, Campillo C, Gov NS, Voituriez R, Azoury J, Umaña-Diaz C, et al. A soft cortex is essential for asymmetric spindle positioning in mouse oocytes. Nat Cell Biol. 2013;15:958–66.CrossRefPubMed
48.
Downes A, Mouras R, Bagnaninchi P, Elfick A. Raman spectroscopy and CARS microscopy of stem cells and their derivatives. J Raman Spectrosc. 2011;42:1864–70.CrossRefPubMedCentralPubMed
Metadata
Title
Evaluation of the impact of vitrification on the actin cytoskeleton of in vitro matured ovine oocytes by means of Raman microspectroscopy
Authors
Luisa Bogliolo
Ombretta Murrone
Massimo Piccinini
Federica Ariu
Sergio Ledda
Sara Tilocca
David F. Albertini
Publication date
01-02-2015
Publisher
Springer US
Published in
Journal of Assisted Reproduction and Genetics / Issue 2/2015
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-014-0389-7

Other articles of this Issue 2/2015

Journal of Assisted Reproduction and Genetics 2/2015 Go to the issue

Genetics

BRCA1 alterations are associated with endometriosis, but BRCA2 alterations show no detectable endometriosis risk: a study in Indian population

Reproductive Physiology and Disease

Effect of induced peritoneal endometriosis on oocyte and embryo quality in a mouse model

Fertility Preservation

The role of menstrual cycle phase and AMH levels in breast cancer patients whose ovarian tissue was cryopreserved for oncofertility treatment

Letter to the Editor

Understanding reproducibility of human IVF traits to predict next IVF cycle outcome; Common mistake

Fertility Preservation

Assessment of vitrification outcome by xenotransplantation of ovarian cortex pieces in γ-irradiated mice: morphological and molecular analyses of apoptosis

Embryo Biology

Administration of the nonsteroidal anti-inflammatory drug tolfenamic acid at embryo transfer improves maintenance of pregnancy and embryo survival in recipient mice