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Gene expression profiling of pluripotency and differentiation-related markers in cat oocytes and preimplantation embryos

Muriel Filliers UGent, Karen Goossens UGent, Ann Van Soom UGent, Barbara Merlo, Charles Earle Pope, Hilde De Rooster UGent, Katrien Smits UGent, Leen Vandaele UGent and Luc Peelman UGent (2012) REPRODUCTION FERTILITY AND DEVELOPMENT. 24(5). p.691-703
abstract
During mammalian preimplantation development, two successive differentiation events lead to the establishment of three committed lineages with separate fates: the trophectoderm, the primitive endoderm and the pluripotent epiblast. In the mouse embryo, the molecular mechanisms underlying these two cell fate decisions have been studied extensively, leading to the identification of lineage-specific transcription factors. Species-specific differences in expression patterns of key regulatory genes have been reported, raising questions regarding their role in different species. The aim of the present study was to characterise the gene expression patterns of pluripotency (OCT4, SOX2, NANOG) and differentiation (CDX2, GATA6)-related markers during feline early development using reverse transcription-quantitative polymerase chain reaction. In addition, we assessed the impact of in vitro development on gene expression by comparing transcript levels of the genes investigated between in vitro and in vivo blastocysts. To normalise quantitative data within different preimplantation embryo stages, we first validated a set of stable reference genes. Transcript levels of all genes investigated were present and changed over the course of preimplantation development; a highly significant embryo-stage effect on gene expression was observed. Transcript levels of OCT4 were significantly reduced in in vitro blastocysts compared with their in vivo counterparts. None of the other genes investigated showed altered expression under in vitro conditions. The different gene expression patterns of OCT4, SOX2, CDX2 and GATA6 in cat embryos resembled those described in mouse embryos, indicative of a preserved role for these genes during early segregation. However, because of the absence of any upregulation of NANOG transcription levels after embryonic genome activation, it is unlikely that NANOG is a key regular of lineage segregation. Such results support the hypothesis that the behaviour of early lineage markers can be species specific. The present study also revealed a pool of maternal NANOG mRNA transcripts, the role of which remains to be elucidated. Comparing transcription levels of these genes between in vivo and in vitro blastocysts revealed low levels of OCT4 mRNA in the latter, which may contribute to the reduced developmental competence of embryos under suboptimal conditions.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
IN-VITRO FERTILIZATION, MESSENGER-RNA EXPRESSION, EARLY MOUSE EMBRYO, OCT-4 EXPRESSION, PRIMITIVE ENDODERM, ES CELLS, BOVINE EMBRYOS, DOMESTIC CAT, EARLY LINEAGE SEGREGATION, OCT4, in vivo blastocyst, early lineage segregation, STEM-CELL LINES
journal title
REPRODUCTION FERTILITY AND DEVELOPMENT
Reprod. Fertil. Dev.
volume
24
issue
5
pages
691 - 703
Web of Science type
Article
Web of Science id
000304603300006
JCR category
ZOOLOGY
JCR impact factor
2.583 (2012)
JCR rank
18/149 (2012)
JCR quartile
1 (2012)
ISSN
1031-3613
DOI
10.1071/RD11068
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
2938669
handle
http://hdl.handle.net/1854/LU-2938669
date created
2012-06-26 16:01:29
date last changed
2012-07-05 14:37:43
@article{2938669,
  abstract     = {During mammalian preimplantation development, two successive differentiation events lead to the establishment of three committed lineages with separate fates: the trophectoderm, the primitive endoderm and the pluripotent epiblast. In the mouse embryo, the molecular mechanisms underlying these two cell fate decisions have been studied extensively, leading to the identification of lineage-specific transcription factors. Species-specific differences in expression patterns of key regulatory genes have been reported, raising questions regarding their role in different species. The aim of the present study was to characterise the gene expression patterns of pluripotency (OCT4, SOX2, NANOG) and differentiation (CDX2, GATA6)-related markers during feline early development using reverse transcription-quantitative polymerase chain reaction. In addition, we assessed the impact of in vitro development on gene expression by comparing transcript levels of the genes investigated between in vitro and in vivo blastocysts. To normalise quantitative data within different preimplantation embryo stages, we first validated a set of stable reference genes. Transcript levels of all genes investigated were present and changed over the course of preimplantation development; a highly significant embryo-stage effect on gene expression was observed. Transcript levels of OCT4 were significantly reduced in in vitro blastocysts compared with their in vivo counterparts. None of the other genes investigated showed altered expression under in vitro conditions. The different gene expression patterns of OCT4, SOX2, CDX2 and GATA6 in cat embryos resembled those described in mouse embryos, indicative of a preserved role for these genes during early segregation. However, because of the absence of any upregulation of NANOG transcription levels after embryonic genome activation, it is unlikely that NANOG is a key regular of lineage segregation. Such results support the hypothesis that the behaviour of early lineage markers can be species specific. The present study also revealed a pool of maternal NANOG mRNA transcripts, the role of which remains to be elucidated. Comparing transcription levels of these genes between in vivo and in vitro blastocysts revealed low levels of OCT4 mRNA in the latter, which may contribute to the reduced developmental competence of embryos under suboptimal conditions.},
  author       = {Filliers, Muriel and Goossens, Karen and Van Soom, Ann and Merlo, Barbara and Pope, Charles Earle and De Rooster, Hilde and Smits, Katrien and Vandaele, Leen and Peelman, Luc},
  issn         = {1031-3613},
  journal      = {REPRODUCTION FERTILITY AND DEVELOPMENT},
  keyword      = {IN-VITRO FERTILIZATION,MESSENGER-RNA EXPRESSION,EARLY MOUSE EMBRYO,OCT-4 EXPRESSION,PRIMITIVE ENDODERM,ES CELLS,BOVINE EMBRYOS,DOMESTIC CAT,EARLY LINEAGE SEGREGATION,OCT4,in vivo blastocyst,early lineage segregation,STEM-CELL LINES},
  language     = {eng},
  number       = {5},
  pages        = {691--703},
  title        = {Gene expression profiling of pluripotency and differentiation-related markers in cat oocytes and preimplantation embryos},
  url          = {http://dx.doi.org/10.1071/RD11068},
  volume       = {24},
  year         = {2012},
}

Chicago
Filliers, Muriel, Karen Goossens, Ann Van Soom, Barbara Merlo, Charles Earle Pope, Hilde De Rooster, Katrien Smits, Leen Vandaele, and Luc Peelman. 2012. “Gene Expression Profiling of Pluripotency and Differentiation-related Markers in Cat Oocytes and Preimplantation Embryos.” Reproduction Fertility and Development 24 (5): 691–703.
APA
Filliers, M., Goossens, K., Van Soom, A., Merlo, B., Pope, C. E., De Rooster, H., Smits, K., et al. (2012). Gene expression profiling of pluripotency and differentiation-related markers in cat oocytes and preimplantation embryos. REPRODUCTION FERTILITY AND DEVELOPMENT, 24(5), 691–703.
Vancouver
1.
Filliers M, Goossens K, Van Soom A, Merlo B, Pope CE, De Rooster H, et al. Gene expression profiling of pluripotency and differentiation-related markers in cat oocytes and preimplantation embryos. REPRODUCTION FERTILITY AND DEVELOPMENT. 2012;24(5):691–703.
MLA
Filliers, Muriel, Karen Goossens, Ann Van Soom, et al. “Gene Expression Profiling of Pluripotency and Differentiation-related Markers in Cat Oocytes and Preimplantation Embryos.” REPRODUCTION FERTILITY AND DEVELOPMENT 24.5 (2012): 691–703. Print.