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SUMO-specific protease 2 is essential for modulating p53-Mdm2 in development of trophoblast stem cell niches and lineages.

Chiu SY, Asai N, Costantini F, Hsu W - PLoS Biol. (2008)

Bottom Line: Reintroducing SENP2 into the mutants can reduce the sumoylation of Mdm2, diminish the p53 level and promote trophoblast development.Furthermore, downregulation of p53 alleviates the SENP2- phenotypes and stimulation of p53 causes abnormalities in trophoblast proliferation and differentiation, resembling those of the SENP2 mutants.Our data reveal a key genetic pathway, SENP2-Mdm2-p53, underlying trophoblast lineage development, suggesting its pivotal role in cell cycle progression of mitosis and endoreduplication.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Genetics, Center for Oral Biology, James P Wilmot Cancer Center, University of Rochester Medical Center, Rochester, New York, USA.

ABSTRACT
SUMO-specific protease 2 (SENP2) modifies proteins by removing SUMO from its substrates. Although SUMO-specific proteases are known to reverse sumoylation in many defined systems, their importance in mammalian development and pathogenesis remains largely elusive. Here we report that SENP2 is highly expressed in trophoblast cells that are required for placentation. Targeted disruption of SENP2 in mice reveals its essential role in development of all three trophoblast layers. The mutation causes a deficiency in cell cycle progression. SENP2 has a specific role in the G-S transition, which is required for mitotic and endoreduplication cell cycles in trophoblast proliferation and differentiation, respectively. SENP2 ablation disturbs the p53-Mdm2 pathway, affecting the expansion of trophoblast progenitors and their maturation. Reintroducing SENP2 into the mutants can reduce the sumoylation of Mdm2, diminish the p53 level and promote trophoblast development. Furthermore, downregulation of p53 alleviates the SENP2- phenotypes and stimulation of p53 causes abnormalities in trophoblast proliferation and differentiation, resembling those of the SENP2 mutants. Our data reveal a key genetic pathway, SENP2-Mdm2-p53, underlying trophoblast lineage development, suggesting its pivotal role in cell cycle progression of mitosis and endoreduplication.

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In Vitro Differentiation of SENP2-Null Blastocysts into Trophoblast Cells Is Defective(A–H) Isolated SENP2+/+ (A) and SENP2–/– (B) blastocysts were cultured for trophoblast differentiation in vitro. Images were taken at culturing day 1 (A,B), day 3 (C,D) and day 6 (E–H). TS cells, outgrowing from the trophectoderm, differentiated into a single trophoblast cell (TC) layer, whereas the ICM formed aggregates and sat on top of the trophoblast cells (C,D).Arrows indicated TGCs present in the cultures (E,F). The cultures were then analyzed by immunostaining of a trophoblast specific marker p450scc (brown) and counterstaining of hematoxylin (blue) on day 6 (G,H).(I) The graph represents the average number of TGC present in the SENP2+/+ and SENP2–/– cultures (p = 0.005, n = 6).Scale bars, 200 μm (E–H); 100 μm (C,D); 50 μm (A,B).
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pbio-0060310-g005: In Vitro Differentiation of SENP2-Null Blastocysts into Trophoblast Cells Is Defective(A–H) Isolated SENP2+/+ (A) and SENP2–/– (B) blastocysts were cultured for trophoblast differentiation in vitro. Images were taken at culturing day 1 (A,B), day 3 (C,D) and day 6 (E–H). TS cells, outgrowing from the trophectoderm, differentiated into a single trophoblast cell (TC) layer, whereas the ICM formed aggregates and sat on top of the trophoblast cells (C,D).Arrows indicated TGCs present in the cultures (E,F). The cultures were then analyzed by immunostaining of a trophoblast specific marker p450scc (brown) and counterstaining of hematoxylin (blue) on day 6 (G,H).(I) The graph represents the average number of TGC present in the SENP2+/+ and SENP2–/– cultures (p = 0.005, n = 6).Scale bars, 200 μm (E–H); 100 μm (C,D); 50 μm (A,B).

Mentions: The abnormal development of TGC caused by SENP2 deficiency was further tested using an in vitro differentiation analysis. The SENP2+/+ and SENP2–/– blastocysts were isolated at E3.5, and cultured to induce TGC differentiation. TS cells growing out from the trophectoderm soon attached to the cultured plates, differentiated, and formed a single trophoblast layer. No noticeable difference was observed between the SENP2+/+ and SENP2–/– blastocysts before hatching (Figure 5A and 5B). About equal amounts of ICM and trophoblast cells developed after 3 d in culture (Figure 5C and 5D). However, although the differentiated TGCs were evident in the SENP2+/+ cultures, their number was significantly reduced in the SENP2–/– cultures after 6 d (Figure 5E–5H). The average number of TGC dropped from 40 in the SENP2+/+ culture to 15 in the SENP2–/– (Figure 5I, p = 0.005, n = 6). Consistent with our in vivo findings, these data suggest that TGC differentiation is severely affected by the loss of SENP2. The results suggest an essential role for SENP2 in TGC development during early placentation.


SUMO-specific protease 2 is essential for modulating p53-Mdm2 in development of trophoblast stem cell niches and lineages.

Chiu SY, Asai N, Costantini F, Hsu W - PLoS Biol. (2008)

In Vitro Differentiation of SENP2-Null Blastocysts into Trophoblast Cells Is Defective(A–H) Isolated SENP2+/+ (A) and SENP2–/– (B) blastocysts were cultured for trophoblast differentiation in vitro. Images were taken at culturing day 1 (A,B), day 3 (C,D) and day 6 (E–H). TS cells, outgrowing from the trophectoderm, differentiated into a single trophoblast cell (TC) layer, whereas the ICM formed aggregates and sat on top of the trophoblast cells (C,D).Arrows indicated TGCs present in the cultures (E,F). The cultures were then analyzed by immunostaining of a trophoblast specific marker p450scc (brown) and counterstaining of hematoxylin (blue) on day 6 (G,H).(I) The graph represents the average number of TGC present in the SENP2+/+ and SENP2–/– cultures (p = 0.005, n = 6).Scale bars, 200 μm (E–H); 100 μm (C,D); 50 μm (A,B).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2602722&req=5

pbio-0060310-g005: In Vitro Differentiation of SENP2-Null Blastocysts into Trophoblast Cells Is Defective(A–H) Isolated SENP2+/+ (A) and SENP2–/– (B) blastocysts were cultured for trophoblast differentiation in vitro. Images were taken at culturing day 1 (A,B), day 3 (C,D) and day 6 (E–H). TS cells, outgrowing from the trophectoderm, differentiated into a single trophoblast cell (TC) layer, whereas the ICM formed aggregates and sat on top of the trophoblast cells (C,D).Arrows indicated TGCs present in the cultures (E,F). The cultures were then analyzed by immunostaining of a trophoblast specific marker p450scc (brown) and counterstaining of hematoxylin (blue) on day 6 (G,H).(I) The graph represents the average number of TGC present in the SENP2+/+ and SENP2–/– cultures (p = 0.005, n = 6).Scale bars, 200 μm (E–H); 100 μm (C,D); 50 μm (A,B).
Mentions: The abnormal development of TGC caused by SENP2 deficiency was further tested using an in vitro differentiation analysis. The SENP2+/+ and SENP2–/– blastocysts were isolated at E3.5, and cultured to induce TGC differentiation. TS cells growing out from the trophectoderm soon attached to the cultured plates, differentiated, and formed a single trophoblast layer. No noticeable difference was observed between the SENP2+/+ and SENP2–/– blastocysts before hatching (Figure 5A and 5B). About equal amounts of ICM and trophoblast cells developed after 3 d in culture (Figure 5C and 5D). However, although the differentiated TGCs were evident in the SENP2+/+ cultures, their number was significantly reduced in the SENP2–/– cultures after 6 d (Figure 5E–5H). The average number of TGC dropped from 40 in the SENP2+/+ culture to 15 in the SENP2–/– (Figure 5I, p = 0.005, n = 6). Consistent with our in vivo findings, these data suggest that TGC differentiation is severely affected by the loss of SENP2. The results suggest an essential role for SENP2 in TGC development during early placentation.

Bottom Line: Reintroducing SENP2 into the mutants can reduce the sumoylation of Mdm2, diminish the p53 level and promote trophoblast development.Furthermore, downregulation of p53 alleviates the SENP2- phenotypes and stimulation of p53 causes abnormalities in trophoblast proliferation and differentiation, resembling those of the SENP2 mutants.Our data reveal a key genetic pathway, SENP2-Mdm2-p53, underlying trophoblast lineage development, suggesting its pivotal role in cell cycle progression of mitosis and endoreduplication.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Genetics, Center for Oral Biology, James P Wilmot Cancer Center, University of Rochester Medical Center, Rochester, New York, USA.

ABSTRACT
SUMO-specific protease 2 (SENP2) modifies proteins by removing SUMO from its substrates. Although SUMO-specific proteases are known to reverse sumoylation in many defined systems, their importance in mammalian development and pathogenesis remains largely elusive. Here we report that SENP2 is highly expressed in trophoblast cells that are required for placentation. Targeted disruption of SENP2 in mice reveals its essential role in development of all three trophoblast layers. The mutation causes a deficiency in cell cycle progression. SENP2 has a specific role in the G-S transition, which is required for mitotic and endoreduplication cell cycles in trophoblast proliferation and differentiation, respectively. SENP2 ablation disturbs the p53-Mdm2 pathway, affecting the expansion of trophoblast progenitors and their maturation. Reintroducing SENP2 into the mutants can reduce the sumoylation of Mdm2, diminish the p53 level and promote trophoblast development. Furthermore, downregulation of p53 alleviates the SENP2- phenotypes and stimulation of p53 causes abnormalities in trophoblast proliferation and differentiation, resembling those of the SENP2 mutants. Our data reveal a key genetic pathway, SENP2-Mdm2-p53, underlying trophoblast lineage development, suggesting its pivotal role in cell cycle progression of mitosis and endoreduplication.

Show MeSH
Related in: MedlinePlus