Limits...
Vascular Notch proteins and Notch signaling in the peri-implantation mouse uterus.

Shawber CJ, Lin L, Gnarra M, Sauer MV, Papaioannou VE, Kitajewski JK, Douglas NC - (2015)

Bottom Line: In the current study, we define the vasculature, expression of Notch proteins and Notch ligands, and Notch activity in both endothelial cells and vascular-associated mural cells of blood vessels in the pre-implantation endometrium and post-implantation decidua of the mouse uterus.Jagged1 is expressed in endothelial cells of spiral arteries and a subset of decidual pericytes.Notch proteins are not expressed in lymphatic vessels or macrophages in the peri-implantation uterus.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Division of Reproductive Sciences, College of Physicians and Surgeons, Columbia University Medical Center, 630 West 168th St, New York, NY 10032 USA ; Department of Surgery, College of Physicians and Surgeons, Columbia University Medical Center, 630 West 168th St, New York, NY 10032 USA.

ABSTRACT

Background: Angiogenesis is essential for uterine decidualization, the progesterone-mediated transformation of the uterus allowing embryo implantation and initiation of pregnancy. In the current study, we define the vasculature, expression of Notch proteins and Notch ligands, and Notch activity in both endothelial cells and vascular-associated mural cells of blood vessels in the pre-implantation endometrium and post-implantation decidua of the mouse uterus.

Methods: We used immunofluorescence to determine the expression of Notch in endothelial cells and mural cells by co-staining for the endothelial cell marker, CD31, the pan-mural cell marker, platelet-derived growth factor receptor beta (PDGFR-β), the pericyte markers, neural/glial antigen 2 (NG2) and desmin, or the smooth muscle cell marker, alpha smooth muscle actin (SMA). A fluorescein isothiocyanate-labeled dextran tracer, was used to identify functional peri-implantation vasculature. CBF:H2B-Venus Notch reporter transgenic mice were used to determine Notch activity.

Results: Notch signaling is observed in endothelial cells and pericytes in the peri-implantation uterus. Prior to implantation, Notch1, Notch2 and Notch4 and Notch ligand, Delta-like 4 (Dll4) are expressed in capillary endothelial cells, while Notch3 is expressed in the pericytes. Jagged1 is expressed in both capillary endothelial cells and pericytes. After implantation, Notch1, Notch4 and Dll4 are expressed in endothelial cells of newly formed decidual capillaries. Jagged1 is expressed in endothelial cells of spiral arteries and a subset of decidual pericytes. Notch proteins are not expressed in lymphatic vessels or macrophages in the peri-implantation uterus.

Conclusions: We show Notch activity and distinct expression patterns for Notch proteins and ligands, suggesting unique roles for Notch1, Notch4, Dll4, and Jag1 during decidual angiogenesis and early placentation. These data set the stage for loss-of-function and gain-of-function studies that will determine the cell-type specific requirements for Notch proteins in decidual angiogenesis and placentation.

No MeSH data available.


Related in: MedlinePlus

Distribution of endothelial cells and patent vessels identified by FITC-dextran in the post-implantation uterus. H&E, IHC and fluorescently labeled dextran in E6.5 uterine sections. (A) H&E at E6.5 highlighting the embryo (e) and inter-implantation sites (i). (B) CD31+ ECs are abundant in the decidua and myometrium. (C) H&E at E6.5 highlighting the embryo surrounded by blood filled maternal sinusoids (box 1, C1) and the blood filled lateral vascular sinuses in the mesometrial decidua (box 2, C2). (D) FITC-dextran is detected in capillaries (D3, arrow) and has extravasated into sinusoids around the embryo (D1), lateral vascular sinuses in the mesometrial decidua (D2 and D3, arrowhead), and the stroma (D3, asterisk). DAPI identifies all nuclei in IF images. am, anti-mesometrial; m, mesometrial; s, serosa. Bar in A and B = 100 μm. Bar in C and D = 500 μm. Bar in A1 and B1, C1, C2, D1 – D3 = 50 μm.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4666149&req=5

Fig3: Distribution of endothelial cells and patent vessels identified by FITC-dextran in the post-implantation uterus. H&E, IHC and fluorescently labeled dextran in E6.5 uterine sections. (A) H&E at E6.5 highlighting the embryo (e) and inter-implantation sites (i). (B) CD31+ ECs are abundant in the decidua and myometrium. (C) H&E at E6.5 highlighting the embryo surrounded by blood filled maternal sinusoids (box 1, C1) and the blood filled lateral vascular sinuses in the mesometrial decidua (box 2, C2). (D) FITC-dextran is detected in capillaries (D3, arrow) and has extravasated into sinusoids around the embryo (D1), lateral vascular sinuses in the mesometrial decidua (D2 and D3, arrowhead), and the stroma (D3, asterisk). DAPI identifies all nuclei in IF images. am, anti-mesometrial; m, mesometrial; s, serosa. Bar in A and B = 100 μm. Bar in C and D = 500 μm. Bar in A1 and B1, C1, C2, D1 – D3 = 50 μm.

Mentions: To determine the presence of functional decidual vessels, we performed FITC-conjugated 10 kDa dextran studies in E6.5 females. FITC-dextran identifies patent capillaries within the mesometrial decidua running laterally from the inter-implantation sites to the embryo (Fig. 3D, D2 and D3). These mesometrial vessels appear to be leaky as areas of extravasation are readily observed. FITC-dextran extravasation is seen into blood filled sinusoids in the implantation crypt around the embryo (Fig. 3D, D1), where CD31 staining is also present (Fig. 3D, B1), and in the lateral vascular sinuses (Fig. 3D, D2 and D3). The distribution of FITC-dextran suggests that functional, patent vessels as well as areas of increased vascular permeability are present in the mesometrial decidua, the site of placenta development.Fig. 3


Vascular Notch proteins and Notch signaling in the peri-implantation mouse uterus.

Shawber CJ, Lin L, Gnarra M, Sauer MV, Papaioannou VE, Kitajewski JK, Douglas NC - (2015)

Distribution of endothelial cells and patent vessels identified by FITC-dextran in the post-implantation uterus. H&E, IHC and fluorescently labeled dextran in E6.5 uterine sections. (A) H&E at E6.5 highlighting the embryo (e) and inter-implantation sites (i). (B) CD31+ ECs are abundant in the decidua and myometrium. (C) H&E at E6.5 highlighting the embryo surrounded by blood filled maternal sinusoids (box 1, C1) and the blood filled lateral vascular sinuses in the mesometrial decidua (box 2, C2). (D) FITC-dextran is detected in capillaries (D3, arrow) and has extravasated into sinusoids around the embryo (D1), lateral vascular sinuses in the mesometrial decidua (D2 and D3, arrowhead), and the stroma (D3, asterisk). DAPI identifies all nuclei in IF images. am, anti-mesometrial; m, mesometrial; s, serosa. Bar in A and B = 100 μm. Bar in C and D = 500 μm. Bar in A1 and B1, C1, C2, D1 – D3 = 50 μm.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4666149&req=5

Fig3: Distribution of endothelial cells and patent vessels identified by FITC-dextran in the post-implantation uterus. H&E, IHC and fluorescently labeled dextran in E6.5 uterine sections. (A) H&E at E6.5 highlighting the embryo (e) and inter-implantation sites (i). (B) CD31+ ECs are abundant in the decidua and myometrium. (C) H&E at E6.5 highlighting the embryo surrounded by blood filled maternal sinusoids (box 1, C1) and the blood filled lateral vascular sinuses in the mesometrial decidua (box 2, C2). (D) FITC-dextran is detected in capillaries (D3, arrow) and has extravasated into sinusoids around the embryo (D1), lateral vascular sinuses in the mesometrial decidua (D2 and D3, arrowhead), and the stroma (D3, asterisk). DAPI identifies all nuclei in IF images. am, anti-mesometrial; m, mesometrial; s, serosa. Bar in A and B = 100 μm. Bar in C and D = 500 μm. Bar in A1 and B1, C1, C2, D1 – D3 = 50 μm.
Mentions: To determine the presence of functional decidual vessels, we performed FITC-conjugated 10 kDa dextran studies in E6.5 females. FITC-dextran identifies patent capillaries within the mesometrial decidua running laterally from the inter-implantation sites to the embryo (Fig. 3D, D2 and D3). These mesometrial vessels appear to be leaky as areas of extravasation are readily observed. FITC-dextran extravasation is seen into blood filled sinusoids in the implantation crypt around the embryo (Fig. 3D, D1), where CD31 staining is also present (Fig. 3D, B1), and in the lateral vascular sinuses (Fig. 3D, D2 and D3). The distribution of FITC-dextran suggests that functional, patent vessels as well as areas of increased vascular permeability are present in the mesometrial decidua, the site of placenta development.Fig. 3

Bottom Line: In the current study, we define the vasculature, expression of Notch proteins and Notch ligands, and Notch activity in both endothelial cells and vascular-associated mural cells of blood vessels in the pre-implantation endometrium and post-implantation decidua of the mouse uterus.Jagged1 is expressed in endothelial cells of spiral arteries and a subset of decidual pericytes.Notch proteins are not expressed in lymphatic vessels or macrophages in the peri-implantation uterus.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Division of Reproductive Sciences, College of Physicians and Surgeons, Columbia University Medical Center, 630 West 168th St, New York, NY 10032 USA ; Department of Surgery, College of Physicians and Surgeons, Columbia University Medical Center, 630 West 168th St, New York, NY 10032 USA.

ABSTRACT

Background: Angiogenesis is essential for uterine decidualization, the progesterone-mediated transformation of the uterus allowing embryo implantation and initiation of pregnancy. In the current study, we define the vasculature, expression of Notch proteins and Notch ligands, and Notch activity in both endothelial cells and vascular-associated mural cells of blood vessels in the pre-implantation endometrium and post-implantation decidua of the mouse uterus.

Methods: We used immunofluorescence to determine the expression of Notch in endothelial cells and mural cells by co-staining for the endothelial cell marker, CD31, the pan-mural cell marker, platelet-derived growth factor receptor beta (PDGFR-β), the pericyte markers, neural/glial antigen 2 (NG2) and desmin, or the smooth muscle cell marker, alpha smooth muscle actin (SMA). A fluorescein isothiocyanate-labeled dextran tracer, was used to identify functional peri-implantation vasculature. CBF:H2B-Venus Notch reporter transgenic mice were used to determine Notch activity.

Results: Notch signaling is observed in endothelial cells and pericytes in the peri-implantation uterus. Prior to implantation, Notch1, Notch2 and Notch4 and Notch ligand, Delta-like 4 (Dll4) are expressed in capillary endothelial cells, while Notch3 is expressed in the pericytes. Jagged1 is expressed in both capillary endothelial cells and pericytes. After implantation, Notch1, Notch4 and Dll4 are expressed in endothelial cells of newly formed decidual capillaries. Jagged1 is expressed in endothelial cells of spiral arteries and a subset of decidual pericytes. Notch proteins are not expressed in lymphatic vessels or macrophages in the peri-implantation uterus.

Conclusions: We show Notch activity and distinct expression patterns for Notch proteins and ligands, suggesting unique roles for Notch1, Notch4, Dll4, and Jag1 during decidual angiogenesis and early placentation. These data set the stage for loss-of-function and gain-of-function studies that will determine the cell-type specific requirements for Notch proteins in decidual angiogenesis and placentation.

No MeSH data available.


Related in: MedlinePlus