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Molecular characterization of EG-VEGF-mediated angiogenesis: differential effects on microvascular and macrovascular endothelial cells.

Brouillet S, Hoffmann P, Benharouga M, Salomon A, Schaal JP, Feige JJ, Alfaidy N - Mol. Biol. Cell (2010)

Bottom Line: Here we characterized its angiogenic effect using different experimental procedures.More importantly, we demonstrated that PROKR1 mediates EG-VEGF angiogenic effects, whereas PROKR2 mediates cellular permeability.Altogether, these data characterized angiogenic processes mediated by EG-VEGF, depicted a new angiogenic factor in the placenta, and suggest a novel view of the regulation of angiogenesis in placental pathologies.

View Article: PubMed Central - PubMed

Affiliation: Institut National de la Santé et de la Recherche Médicale, Unité 878, Grenoble, France.

ABSTRACT
Endocrine gland derived vascular endothelial growth factor (EG-VEGF) also called prokineticin (PK1), has been identified and linked to several biological processes including angiogenesis. EG-VEGF is abundantly expressed in the highest vascularized organ, the human placenta. Here we characterized its angiogenic effect using different experimental procedures. Immunohistochemistry was used to localize EG-VEGF receptors (PROKR1 and PROKR2) in placental and umbilical cord tissue. Primary microvascular placental endothelial cell (HPEC) and umbilical vein-derived macrovascular EC (HUVEC) were used to assess its effects on proliferation, migration, cell survival, pseudovascular organization, spheroid sprouting, permeability and paracellular transport. siRNA and neutralizing antibody strategies were used to differentiate PROKR1- from PROKR2-mediated effects. Our results show that 1) HPEC and HUVEC express both types of receptors 2) EG-VEGF stimulates HPEC's proliferation, migration and survival, but increases only survival in HUVECs. and 3) EG-VEGF was more potent than VEGF in stimulating HPEC sprout formation, pseudovascular organization, and it significantly increases HPEC permeability and paracellular transport. More importantly, we demonstrated that PROKR1 mediates EG-VEGF angiogenic effects, whereas PROKR2 mediates cellular permeability. Altogether, these data characterized angiogenic processes mediated by EG-VEGF, depicted a new angiogenic factor in the placenta, and suggest a novel view of the regulation of angiogenesis in placental pathologies.

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EG-VEGF increases HPEC but not HUVEC cord-like organization. (A) Photographs of HPEC and HUVEC cells cultured on Matrigel for 0 and 10 h in the absence or the presence of EG-VEGF (25 ng/ml). Note that EG-VEGF increased HPEC but not HUVEC organization into cord-like structures compared with the control condition. (B) Measurements of the number of branches formed by the cells after 10 h of culture in the absence or presence of EG-VEGF. *p < 0.05.
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Figure 4: EG-VEGF increases HPEC but not HUVEC cord-like organization. (A) Photographs of HPEC and HUVEC cells cultured on Matrigel for 0 and 10 h in the absence or the presence of EG-VEGF (25 ng/ml). Note that EG-VEGF increased HPEC but not HUVEC organization into cord-like structures compared with the control condition. (B) Measurements of the number of branches formed by the cells after 10 h of culture in the absence or presence of EG-VEGF. *p < 0.05.

Mentions: Endothelial cells are well known to self-organize as networks of vascular-like structures when grown on Matrigel (Murray, 2003). Here, we investigated the effect of EG-VEGF on pseudovascular organization of HPECs and HUVECs, with photographs taken every hour. Figure 4A shows representative photographs of HPEC and HUVEC cells at 0 and 10 h of culture on Matrigel and under different treatments. In the control condition, HPEC start to organize into tube-like structures by 3 h. By 10 h of culture only half of the plate was organized in a network of tubular structures. In the EG-VEGF treatment condition, this process was increased with an organization starting as early as after 1 h of culture, and the plate was completely organized by 10 h of culture. In contrast, EG-VEGF did not affect HUVEC organization. Quantification of four independent experiments shows that EG-VEGF significantly increased HPEC but not HUVEC organization as compared with the control condition, (Figure 4B).


Molecular characterization of EG-VEGF-mediated angiogenesis: differential effects on microvascular and macrovascular endothelial cells.

Brouillet S, Hoffmann P, Benharouga M, Salomon A, Schaal JP, Feige JJ, Alfaidy N - Mol. Biol. Cell (2010)

EG-VEGF increases HPEC but not HUVEC cord-like organization. (A) Photographs of HPEC and HUVEC cells cultured on Matrigel for 0 and 10 h in the absence or the presence of EG-VEGF (25 ng/ml). Note that EG-VEGF increased HPEC but not HUVEC organization into cord-like structures compared with the control condition. (B) Measurements of the number of branches formed by the cells after 10 h of culture in the absence or presence of EG-VEGF. *p < 0.05.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2921113&req=5

Figure 4: EG-VEGF increases HPEC but not HUVEC cord-like organization. (A) Photographs of HPEC and HUVEC cells cultured on Matrigel for 0 and 10 h in the absence or the presence of EG-VEGF (25 ng/ml). Note that EG-VEGF increased HPEC but not HUVEC organization into cord-like structures compared with the control condition. (B) Measurements of the number of branches formed by the cells after 10 h of culture in the absence or presence of EG-VEGF. *p < 0.05.
Mentions: Endothelial cells are well known to self-organize as networks of vascular-like structures when grown on Matrigel (Murray, 2003). Here, we investigated the effect of EG-VEGF on pseudovascular organization of HPECs and HUVECs, with photographs taken every hour. Figure 4A shows representative photographs of HPEC and HUVEC cells at 0 and 10 h of culture on Matrigel and under different treatments. In the control condition, HPEC start to organize into tube-like structures by 3 h. By 10 h of culture only half of the plate was organized in a network of tubular structures. In the EG-VEGF treatment condition, this process was increased with an organization starting as early as after 1 h of culture, and the plate was completely organized by 10 h of culture. In contrast, EG-VEGF did not affect HUVEC organization. Quantification of four independent experiments shows that EG-VEGF significantly increased HPEC but not HUVEC organization as compared with the control condition, (Figure 4B).

Bottom Line: Here we characterized its angiogenic effect using different experimental procedures.More importantly, we demonstrated that PROKR1 mediates EG-VEGF angiogenic effects, whereas PROKR2 mediates cellular permeability.Altogether, these data characterized angiogenic processes mediated by EG-VEGF, depicted a new angiogenic factor in the placenta, and suggest a novel view of the regulation of angiogenesis in placental pathologies.

View Article: PubMed Central - PubMed

Affiliation: Institut National de la Santé et de la Recherche Médicale, Unité 878, Grenoble, France.

ABSTRACT
Endocrine gland derived vascular endothelial growth factor (EG-VEGF) also called prokineticin (PK1), has been identified and linked to several biological processes including angiogenesis. EG-VEGF is abundantly expressed in the highest vascularized organ, the human placenta. Here we characterized its angiogenic effect using different experimental procedures. Immunohistochemistry was used to localize EG-VEGF receptors (PROKR1 and PROKR2) in placental and umbilical cord tissue. Primary microvascular placental endothelial cell (HPEC) and umbilical vein-derived macrovascular EC (HUVEC) were used to assess its effects on proliferation, migration, cell survival, pseudovascular organization, spheroid sprouting, permeability and paracellular transport. siRNA and neutralizing antibody strategies were used to differentiate PROKR1- from PROKR2-mediated effects. Our results show that 1) HPEC and HUVEC express both types of receptors 2) EG-VEGF stimulates HPEC's proliferation, migration and survival, but increases only survival in HUVECs. and 3) EG-VEGF was more potent than VEGF in stimulating HPEC sprout formation, pseudovascular organization, and it significantly increases HPEC permeability and paracellular transport. More importantly, we demonstrated that PROKR1 mediates EG-VEGF angiogenic effects, whereas PROKR2 mediates cellular permeability. Altogether, these data characterized angiogenic processes mediated by EG-VEGF, depicted a new angiogenic factor in the placenta, and suggest a novel view of the regulation of angiogenesis in placental pathologies.

Show MeSH
Related in: MedlinePlus