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In vitro fertilization and embryo culture strongly impact the placental transcriptome in the mouse model.

Fauque P, Mondon F, Letourneur F, Ripoche MA, Journot L, Barbaux S, Dandolo L, Patrat C, Wolf JP, Jouannet P, Jammes H, Vaiman D - PLoS ONE (2010)

Bottom Line: Imprinted genes were modified similarly to the X.Expression of X and imprinted genes is also greatly modulated probably to adapt to adverse conditions.Our results highlight the importance of studying human placentas from ART.

View Article: PubMed Central - PubMed

Affiliation: Service d'Histologie-Embryologie, Biologie de la Reproduction, Hôpital Cochin, Paris, France. patricia.fauque@cch.aphp.fr

ABSTRACT

Background: Assisted Reproductive Technologies (ART) are increasingly used in humans; however, their impact is now questioned. At blastocyst stage, the trophectoderm is directly in contact with an artificial medium environment, which can impact placental development. This study was designed to carry out an in-depth analysis of the placental transcriptome after ART in mice.

Methodology/principal findings: Blastocysts were transferred either (1) after in vivo fertilization and development (control group) or (2) after in vitro fertilization and embryo culture. Placentas were then analyzed at E10.5. Six percent of transcripts were altered at the two-fold threshold in placentas of manipulated embryos, 2/3 of transcripts being down-regulated. Strikingly, the X-chromosome harbors 11% of altered genes, 2/3 being induced. Imprinted genes were modified similarly to the X. Promoter composition analysis indicates that FOXA transcription factors may be involved in the transcriptional deregulations.

Conclusions: For the first time, our study shows that in vitro fertilization associated with embryo culture strongly modify the placental expression profile, long after embryo manipulations, meaning that the stress of artificial environment is memorized after implantation. Expression of X and imprinted genes is also greatly modulated probably to adapt to adverse conditions. Our results highlight the importance of studying human placentas from ART.

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Hierarchical clustering analysis according to the different procedures (IVF M16 and G1/G2 groups and control group).
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pone-0009218-g003: Hierarchical clustering analysis according to the different procedures (IVF M16 and G1/G2 groups and control group).

Mentions: The expression of placental genes was compared between placentas from the different groups (Figure 1). The Affymetrix 430 2.0 microarray that was used encompasses a complete set of mouse transcripts (45,101 transcripts). We calculated a ratio of modification for each gene, by pairwise comparisons between the three conditions (in vivo, in vitro M16, and in vitro G1/G2), in order to count how many genes were modified at the two-fold or four-fold thresholds (either up- or down-regulated), as shown in Table 1 and Figure 2. At the two-fold threshold, in vitro fertilization and embryo culture in M16 resulted in the overall deregulation of 3,553 transcripts compared to in vivo produced embryos, while G1/G2 culture induced the expressional alterations of 2,721 transcripts. At the four-fold threshold, the number of modified transcripts was reduced more than five times, with 703 and 494 misregulated transcripts after culture in M16 and G1/G2, respectively. Overall, the count of dysregulated transcripts after in vitro fertilization and embryo culture versus in vivo conditions was much higher than when the two in vitro conditions of embryo culture were compared (Table 1). The same idea is illustrated when all the transcripts were taken into consideration by a hierarchical clustering analysis of the expression profiles. This representation shows a striking separation into two major branches opposing the in vivo sample to the two in vitro samples (correlation coefficient between IVF M16 and IVF G1/G2  = 0.67, Figure 3). IVF M16 and IVF G1/G2 samples were further divided into two little branches (correlation coefficient IVF M16 versus control  = 0.14 and IVF G1/G2 versus control  = 0.27).


In vitro fertilization and embryo culture strongly impact the placental transcriptome in the mouse model.

Fauque P, Mondon F, Letourneur F, Ripoche MA, Journot L, Barbaux S, Dandolo L, Patrat C, Wolf JP, Jouannet P, Jammes H, Vaiman D - PLoS ONE (2010)

Hierarchical clustering analysis according to the different procedures (IVF M16 and G1/G2 groups and control group).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2821408&req=5

pone-0009218-g003: Hierarchical clustering analysis according to the different procedures (IVF M16 and G1/G2 groups and control group).
Mentions: The expression of placental genes was compared between placentas from the different groups (Figure 1). The Affymetrix 430 2.0 microarray that was used encompasses a complete set of mouse transcripts (45,101 transcripts). We calculated a ratio of modification for each gene, by pairwise comparisons between the three conditions (in vivo, in vitro M16, and in vitro G1/G2), in order to count how many genes were modified at the two-fold or four-fold thresholds (either up- or down-regulated), as shown in Table 1 and Figure 2. At the two-fold threshold, in vitro fertilization and embryo culture in M16 resulted in the overall deregulation of 3,553 transcripts compared to in vivo produced embryos, while G1/G2 culture induced the expressional alterations of 2,721 transcripts. At the four-fold threshold, the number of modified transcripts was reduced more than five times, with 703 and 494 misregulated transcripts after culture in M16 and G1/G2, respectively. Overall, the count of dysregulated transcripts after in vitro fertilization and embryo culture versus in vivo conditions was much higher than when the two in vitro conditions of embryo culture were compared (Table 1). The same idea is illustrated when all the transcripts were taken into consideration by a hierarchical clustering analysis of the expression profiles. This representation shows a striking separation into two major branches opposing the in vivo sample to the two in vitro samples (correlation coefficient between IVF M16 and IVF G1/G2  = 0.67, Figure 3). IVF M16 and IVF G1/G2 samples were further divided into two little branches (correlation coefficient IVF M16 versus control  = 0.14 and IVF G1/G2 versus control  = 0.27).

Bottom Line: Imprinted genes were modified similarly to the X.Expression of X and imprinted genes is also greatly modulated probably to adapt to adverse conditions.Our results highlight the importance of studying human placentas from ART.

View Article: PubMed Central - PubMed

Affiliation: Service d'Histologie-Embryologie, Biologie de la Reproduction, Hôpital Cochin, Paris, France. patricia.fauque@cch.aphp.fr

ABSTRACT

Background: Assisted Reproductive Technologies (ART) are increasingly used in humans; however, their impact is now questioned. At blastocyst stage, the trophectoderm is directly in contact with an artificial medium environment, which can impact placental development. This study was designed to carry out an in-depth analysis of the placental transcriptome after ART in mice.

Methodology/principal findings: Blastocysts were transferred either (1) after in vivo fertilization and development (control group) or (2) after in vitro fertilization and embryo culture. Placentas were then analyzed at E10.5. Six percent of transcripts were altered at the two-fold threshold in placentas of manipulated embryos, 2/3 of transcripts being down-regulated. Strikingly, the X-chromosome harbors 11% of altered genes, 2/3 being induced. Imprinted genes were modified similarly to the X. Promoter composition analysis indicates that FOXA transcription factors may be involved in the transcriptional deregulations.

Conclusions: For the first time, our study shows that in vitro fertilization associated with embryo culture strongly modify the placental expression profile, long after embryo manipulations, meaning that the stress of artificial environment is memorized after implantation. Expression of X and imprinted genes is also greatly modulated probably to adapt to adverse conditions. Our results highlight the importance of studying human placentas from ART.

Show MeSH