Gene expression profiles of vitrified in vivo derived 8-cell stage mouse embryos detected by high density oligonucleotide microarrays

Solomon Mamo, Szilard Bodo, Julianna Kobolak, Zsuzsanna Polgar, Gergely Tolgyesi, A. Dinnyés

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Very little is known about the effect of vitrification on gene functions after warming. The goals of our study were to compare the gene expression patterns, and identify those most affected. For this, 8-cell stage embryos were collected from ICR mice and vitrified with solid surface vitrification technique, while maintaining equal numbers of embryos as control. Total RNAs were extracted and two rounds of amplification were employed. Finally three micrograms of contrasting RNA samples were hybridized on the Agilent Mouse 22 K oligonucleotide slides and the results were analyzed with subsequent verification by independent real-time PCR analyses. The two rounds of amplification with 5 μg tRNA input have yielded 15-16 μg of cRNA. The analyses of repeated hybridizations showed 20,183 genes/ESTs as common signatures, and unsupervised analysis identified 628 differentially expressed (P <0.01) genes. However, with at least 1.5-fold change considerations, 183 genes were differentially expressed (P <0.01) out of which 107 were upregulated. The independent analysis with real-time PCR and unamplified samples fully confirmed the results of microarray, indicating the linearity of amplification. Furthermore, this novel gene expression study for vitrified embryos identified many new candidate genes with overrepresentation in some important biological processes. Thus, it is possible to conclude that the expression pattern reflected a broad spectrum of consequences of vitrification on embryos, with most effects on metabolism, regulatory role and stress response genes and allowed the identification of new candidate marker genes for cryosurvival.

Original languageEnglish
Pages (from-to)1380-1392
Number of pages13
JournalMolecular Reproduction and Development
Volume73
Issue number11
DOIs
Publication statusPublished - Nov 2006

Fingerprint

Oligonucleotide Array Sequence Analysis
Transcriptome
Embryonic Structures
Vitrification
Genes
Real-Time Polymerase Chain Reaction
RNA
Biological Phenomena
Gene Expression
Complementary RNA
Inbred ICR Mouse
Expressed Sequence Tags
Transfer RNA
Oligonucleotides

Keywords

  • Cryopreservation
  • Gene expression
  • Microarray
  • RNA amplification
  • Vitrification

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology
  • Cell Biology

Cite this

Gene expression profiles of vitrified in vivo derived 8-cell stage mouse embryos detected by high density oligonucleotide microarrays. / Mamo, Solomon; Bodo, Szilard; Kobolak, Julianna; Polgar, Zsuzsanna; Tolgyesi, Gergely; Dinnyés, A.

In: Molecular Reproduction and Development, Vol. 73, No. 11, 11.2006, p. 1380-1392.

Research output: Contribution to journalArticle

Mamo, Solomon ; Bodo, Szilard ; Kobolak, Julianna ; Polgar, Zsuzsanna ; Tolgyesi, Gergely ; Dinnyés, A. / Gene expression profiles of vitrified in vivo derived 8-cell stage mouse embryos detected by high density oligonucleotide microarrays. In: Molecular Reproduction and Development. 2006 ; Vol. 73, No. 11. pp. 1380-1392.
@article{89d31443a69f481e946accae2b43596f,
title = "Gene expression profiles of vitrified in vivo derived 8-cell stage mouse embryos detected by high density oligonucleotide microarrays",
abstract = "Very little is known about the effect of vitrification on gene functions after warming. The goals of our study were to compare the gene expression patterns, and identify those most affected. For this, 8-cell stage embryos were collected from ICR mice and vitrified with solid surface vitrification technique, while maintaining equal numbers of embryos as control. Total RNAs were extracted and two rounds of amplification were employed. Finally three micrograms of contrasting RNA samples were hybridized on the Agilent Mouse 22 K oligonucleotide slides and the results were analyzed with subsequent verification by independent real-time PCR analyses. The two rounds of amplification with 5 μg tRNA input have yielded 15-16 μg of cRNA. The analyses of repeated hybridizations showed 20,183 genes/ESTs as common signatures, and unsupervised analysis identified 628 differentially expressed (P <0.01) genes. However, with at least 1.5-fold change considerations, 183 genes were differentially expressed (P <0.01) out of which 107 were upregulated. The independent analysis with real-time PCR and unamplified samples fully confirmed the results of microarray, indicating the linearity of amplification. Furthermore, this novel gene expression study for vitrified embryos identified many new candidate genes with overrepresentation in some important biological processes. Thus, it is possible to conclude that the expression pattern reflected a broad spectrum of consequences of vitrification on embryos, with most effects on metabolism, regulatory role and stress response genes and allowed the identification of new candidate marker genes for cryosurvival.",
keywords = "Cryopreservation, Gene expression, Microarray, RNA amplification, Vitrification",
author = "Solomon Mamo and Szilard Bodo and Julianna Kobolak and Zsuzsanna Polgar and Gergely Tolgyesi and A. Dinny{\'e}s",
year = "2006",
month = "11",
doi = "10.1002/mrd.20588",
language = "English",
volume = "73",
pages = "1380--1392",
journal = "Molecular Reproduction and Development",
issn = "1040-452X",
publisher = "Wiley-Liss Inc.",
number = "11",

}

TY - JOUR

T1 - Gene expression profiles of vitrified in vivo derived 8-cell stage mouse embryos detected by high density oligonucleotide microarrays

AU - Mamo, Solomon

AU - Bodo, Szilard

AU - Kobolak, Julianna

AU - Polgar, Zsuzsanna

AU - Tolgyesi, Gergely

AU - Dinnyés, A.

PY - 2006/11

Y1 - 2006/11

N2 - Very little is known about the effect of vitrification on gene functions after warming. The goals of our study were to compare the gene expression patterns, and identify those most affected. For this, 8-cell stage embryos were collected from ICR mice and vitrified with solid surface vitrification technique, while maintaining equal numbers of embryos as control. Total RNAs were extracted and two rounds of amplification were employed. Finally three micrograms of contrasting RNA samples were hybridized on the Agilent Mouse 22 K oligonucleotide slides and the results were analyzed with subsequent verification by independent real-time PCR analyses. The two rounds of amplification with 5 μg tRNA input have yielded 15-16 μg of cRNA. The analyses of repeated hybridizations showed 20,183 genes/ESTs as common signatures, and unsupervised analysis identified 628 differentially expressed (P <0.01) genes. However, with at least 1.5-fold change considerations, 183 genes were differentially expressed (P <0.01) out of which 107 were upregulated. The independent analysis with real-time PCR and unamplified samples fully confirmed the results of microarray, indicating the linearity of amplification. Furthermore, this novel gene expression study for vitrified embryos identified many new candidate genes with overrepresentation in some important biological processes. Thus, it is possible to conclude that the expression pattern reflected a broad spectrum of consequences of vitrification on embryos, with most effects on metabolism, regulatory role and stress response genes and allowed the identification of new candidate marker genes for cryosurvival.

AB - Very little is known about the effect of vitrification on gene functions after warming. The goals of our study were to compare the gene expression patterns, and identify those most affected. For this, 8-cell stage embryos were collected from ICR mice and vitrified with solid surface vitrification technique, while maintaining equal numbers of embryos as control. Total RNAs were extracted and two rounds of amplification were employed. Finally three micrograms of contrasting RNA samples were hybridized on the Agilent Mouse 22 K oligonucleotide slides and the results were analyzed with subsequent verification by independent real-time PCR analyses. The two rounds of amplification with 5 μg tRNA input have yielded 15-16 μg of cRNA. The analyses of repeated hybridizations showed 20,183 genes/ESTs as common signatures, and unsupervised analysis identified 628 differentially expressed (P <0.01) genes. However, with at least 1.5-fold change considerations, 183 genes were differentially expressed (P <0.01) out of which 107 were upregulated. The independent analysis with real-time PCR and unamplified samples fully confirmed the results of microarray, indicating the linearity of amplification. Furthermore, this novel gene expression study for vitrified embryos identified many new candidate genes with overrepresentation in some important biological processes. Thus, it is possible to conclude that the expression pattern reflected a broad spectrum of consequences of vitrification on embryos, with most effects on metabolism, regulatory role and stress response genes and allowed the identification of new candidate marker genes for cryosurvival.

KW - Cryopreservation

KW - Gene expression

KW - Microarray

KW - RNA amplification

KW - Vitrification

UR - http://www.scopus.com/inward/record.url?scp=33748849063&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33748849063&partnerID=8YFLogxK

U2 - 10.1002/mrd.20588

DO - 10.1002/mrd.20588

M3 - Article

C2 - 16897732

AN - SCOPUS:33748849063

VL - 73

SP - 1380

EP - 1392

JO - Molecular Reproduction and Development

JF - Molecular Reproduction and Development

SN - 1040-452X

IS - 11

ER -