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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

Notch activity in endothelial cells and mural cells in the peri-implantation uterus. (A) H&E at E3.5 highlighting the luminal epithelium (arrows), inner circular and outer longitudinal myometrium [dashed bracket (myo)] and endometrial stroma (st). (B, C) IF of sections through the uterus of CBF:H2B-Venus Notch reporter mice enabled assessment of Notch activity at E3.5. Ovals indicate areas of the uteri magnified (B1, C1). (B) Notch activity was observed in CD31+ ECs (red) in the stroma (B1), myometrium, and glandular epithelium (arrowheads). (C) Notch activity was observed in PDGFR-β+ mural cells in the stroma (C1) and myometrium. (D) H&E at E6.5 highlighting the embryo (e), inter-implantation sites (i), and myometrium (myo). (E, F) IF of sections through implantation sites of female CBF:H2B-Venus Notch reporter mice at E6.5. Ovals indicate areas of the uteri magnified (E1, F1). (E) Notch activity was observed in CD31+ ECs of decidual capillaries (E1) and myometrial vessels. (F) Notch activity was observed in PDGFR-β+ mural cells in the anti-mesometrial decidua. DAPI identifies all nuclei in IF images. am, anti-mesometrial; m, mesometrial. Bar in A and D =100 μm. Bar in B, C, E, and F = 500 μm. Bar in B1, C1, E1, and F1 = 100 μm.
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Fig11: Notch activity in endothelial cells and mural cells in the peri-implantation uterus. (A) H&E at E3.5 highlighting the luminal epithelium (arrows), inner circular and outer longitudinal myometrium [dashed bracket (myo)] and endometrial stroma (st). (B, C) IF of sections through the uterus of CBF:H2B-Venus Notch reporter mice enabled assessment of Notch activity at E3.5. Ovals indicate areas of the uteri magnified (B1, C1). (B) Notch activity was observed in CD31+ ECs (red) in the stroma (B1), myometrium, and glandular epithelium (arrowheads). (C) Notch activity was observed in PDGFR-β+ mural cells in the stroma (C1) and myometrium. (D) H&E at E6.5 highlighting the embryo (e), inter-implantation sites (i), and myometrium (myo). (E, F) IF of sections through implantation sites of female CBF:H2B-Venus Notch reporter mice at E6.5. Ovals indicate areas of the uteri magnified (E1, F1). (E) Notch activity was observed in CD31+ ECs of decidual capillaries (E1) and myometrial vessels. (F) Notch activity was observed in PDGFR-β+ mural cells in the anti-mesometrial decidua. DAPI identifies all nuclei in IF images. am, anti-mesometrial; m, mesometrial. Bar in A and D =100 μm. Bar in B, C, E, and F = 500 μm. Bar in B1, C1, E1, and F1 = 100 μm.

Mentions: To determine Notch activity in the peri-implantation uterus, female CBF:H2B-Venus Notch reporter mice were mated to wild-type males. YFP expression was used to determine Notch signaling activity in CD31+ decidual ECs and PDGFR-β+ pericytes. At E3.5, Notch activity is observed in CD31+ ECs in the endometrium and myometrium (Fig. 11B, B1), in luminal and glandular epithelial cells (Fig. 11B and C, arrowheads), and in a subset of PDGFR-β+ pericytes (Fig. 11C, C1). At E6.5, the H2B-Venus transgene is expressed in CD31+ ECs of decidual capillaries (Fig. 11E, E1), CD31+ ECs of myometrial vessels (Fig. 11E) and in a subset of PDGFR-β+ pericytes in the anti-mesometrial decidua (Fig. 11F, F1). Thus, Notch activity is observed in ECs and pericytes in the peri-implantation uterus.Fig. 11


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)

Notch activity in endothelial cells and mural cells in the peri-implantation uterus. (A) H&E at E3.5 highlighting the luminal epithelium (arrows), inner circular and outer longitudinal myometrium [dashed bracket (myo)] and endometrial stroma (st). (B, C) IF of sections through the uterus of CBF:H2B-Venus Notch reporter mice enabled assessment of Notch activity at E3.5. Ovals indicate areas of the uteri magnified (B1, C1). (B) Notch activity was observed in CD31+ ECs (red) in the stroma (B1), myometrium, and glandular epithelium (arrowheads). (C) Notch activity was observed in PDGFR-β+ mural cells in the stroma (C1) and myometrium. (D) H&E at E6.5 highlighting the embryo (e), inter-implantation sites (i), and myometrium (myo). (E, F) IF of sections through implantation sites of female CBF:H2B-Venus Notch reporter mice at E6.5. Ovals indicate areas of the uteri magnified (E1, F1). (E) Notch activity was observed in CD31+ ECs of decidual capillaries (E1) and myometrial vessels. (F) Notch activity was observed in PDGFR-β+ mural cells in the anti-mesometrial decidua. DAPI identifies all nuclei in IF images. am, anti-mesometrial; m, mesometrial. Bar in A and D =100 μm. Bar in B, C, E, and F = 500 μm. Bar in B1, C1, E1, and F1 = 100 μm.
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Related In: Results  -  Collection

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Fig11: Notch activity in endothelial cells and mural cells in the peri-implantation uterus. (A) H&E at E3.5 highlighting the luminal epithelium (arrows), inner circular and outer longitudinal myometrium [dashed bracket (myo)] and endometrial stroma (st). (B, C) IF of sections through the uterus of CBF:H2B-Venus Notch reporter mice enabled assessment of Notch activity at E3.5. Ovals indicate areas of the uteri magnified (B1, C1). (B) Notch activity was observed in CD31+ ECs (red) in the stroma (B1), myometrium, and glandular epithelium (arrowheads). (C) Notch activity was observed in PDGFR-β+ mural cells in the stroma (C1) and myometrium. (D) H&E at E6.5 highlighting the embryo (e), inter-implantation sites (i), and myometrium (myo). (E, F) IF of sections through implantation sites of female CBF:H2B-Venus Notch reporter mice at E6.5. Ovals indicate areas of the uteri magnified (E1, F1). (E) Notch activity was observed in CD31+ ECs of decidual capillaries (E1) and myometrial vessels. (F) Notch activity was observed in PDGFR-β+ mural cells in the anti-mesometrial decidua. DAPI identifies all nuclei in IF images. am, anti-mesometrial; m, mesometrial. Bar in A and D =100 μm. Bar in B, C, E, and F = 500 μm. Bar in B1, C1, E1, and F1 = 100 μm.
Mentions: To determine Notch activity in the peri-implantation uterus, female CBF:H2B-Venus Notch reporter mice were mated to wild-type males. YFP expression was used to determine Notch signaling activity in CD31+ decidual ECs and PDGFR-β+ pericytes. At E3.5, Notch activity is observed in CD31+ ECs in the endometrium and myometrium (Fig. 11B, B1), in luminal and glandular epithelial cells (Fig. 11B and C, arrowheads), and in a subset of PDGFR-β+ pericytes (Fig. 11C, C1). At E6.5, the H2B-Venus transgene is expressed in CD31+ ECs of decidual capillaries (Fig. 11E, E1), CD31+ ECs of myometrial vessels (Fig. 11E) and in a subset of PDGFR-β+ pericytes in the anti-mesometrial decidua (Fig. 11F, F1). Thus, Notch activity is observed in ECs and pericytes in the peri-implantation uterus.Fig. 11

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