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Class 3 semaphorins negatively regulate dermal lymphatic network formation.

Uchida Y, James JM, Suto F, Mukouyama YS - Biol Open (2015)

Bottom Line: In contrast, Sema3f;Sema3g double mutants display increased lymphatic branching, while Nrp2 mutants exhibit reduced lymphatic branching.Our results provide the first evidence that SEMA3F and SEMA3G function as a negative regulator for dermal lymphangiogenesis in vivo.The reciprocal phenotype in lymphatic branching between Sema3f;Sema3g double mutants and Nrp2 mutants suggest a complex NRP2 function that regulates LEC behavior both positively and negatively, through a binding with VEGFC or SEMA3s.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Stem Cell and Neuro-Vascular Biology, Genetics and Developmental Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Building 10/6C103, 10 Center Drive, Bethesda, MD 20892, USA.

No MeSH data available.


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Expression of SEMA3s and their receptor NRP2 in limb skin. (A,B) Expression of Sema3c and Sema3f in E15.5 skin (arrows). In situ hybridization on serial transverse sections was performed with the indicated probes. (C,D) Whole-mount limb skin staining of E17.5 Nrp2taugfp/+ embryos with antibodies to the LEC marker PROX1 (red), LYVE1 (blue), and the EC marker PECAM1 (red) together with GFP (Nrp2taugfp, green). A, arteries; L, lymphatic vessels; V, veins. (E,E′) Whole-mount limb skin staining of E17.5 Sema3glacZ/+;Nrp2taugfp/+ embryos with antibodies to β-gal (Sema3glacZ, red), GFP (Nrp2taugfp, green), and PECAM1 (blue). Sema3glacZ is expressed in arteries (A, open arrowhead), whereas Nrp2taugfp is expressed in both lymphatic vessels (L, arrow) and veins (V, arrowhead). (F) Schematic model illustrating differential expression patterns of SEMA3s and NRP2 in the skin. Scale bars: 100 µm.
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BIO012302F2: Expression of SEMA3s and their receptor NRP2 in limb skin. (A,B) Expression of Sema3c and Sema3f in E15.5 skin (arrows). In situ hybridization on serial transverse sections was performed with the indicated probes. (C,D) Whole-mount limb skin staining of E17.5 Nrp2taugfp/+ embryos with antibodies to the LEC marker PROX1 (red), LYVE1 (blue), and the EC marker PECAM1 (red) together with GFP (Nrp2taugfp, green). A, arteries; L, lymphatic vessels; V, veins. (E,E′) Whole-mount limb skin staining of E17.5 Sema3glacZ/+;Nrp2taugfp/+ embryos with antibodies to β-gal (Sema3glacZ, red), GFP (Nrp2taugfp, green), and PECAM1 (blue). Sema3glacZ is expressed in arteries (A, open arrowhead), whereas Nrp2taugfp is expressed in both lymphatic vessels (L, arrow) and veins (V, arrowhead). (F) Schematic model illustrating differential expression patterns of SEMA3s and NRP2 in the skin. Scale bars: 100 µm.

Mentions: We next examined the expression patterns of Sema3b, Sema3c, Sema3f and Sema3g in E15.5 skin by in situ hybridization, stages when lymphatic network is actively developing in the skin. We found distinct expression patterns of Sema3c and Sema3f (Fig. 2A,B), albeit a less clear in situ hybridization signal for Sema3b and Sema3g (data not shown). Expression of Sema3c was detectable in the layer adjacent to the boundary between the dermis and hypodermis (Fig. 2A). Consistent with a previous report (Eckhardt and Meyerhans, 1998), expression of Sema3f was observed in the epidermis (Fig. 2B). Although the Sema3g in situ hybridization signal was not clearly detectable, the expression of SEMA3G using Sema3glacZ/+ heterozygous embryos, carrying a lacZ reporter cassette under the endogenous Sema3g promoter, was restricted to arteries and is not expressed in veins or lymphatic vessels within the skin (Fig. 2E,E′; Kutschera et al., 2011).Fig. 2.


Class 3 semaphorins negatively regulate dermal lymphatic network formation.

Uchida Y, James JM, Suto F, Mukouyama YS - Biol Open (2015)

Expression of SEMA3s and their receptor NRP2 in limb skin. (A,B) Expression of Sema3c and Sema3f in E15.5 skin (arrows). In situ hybridization on serial transverse sections was performed with the indicated probes. (C,D) Whole-mount limb skin staining of E17.5 Nrp2taugfp/+ embryos with antibodies to the LEC marker PROX1 (red), LYVE1 (blue), and the EC marker PECAM1 (red) together with GFP (Nrp2taugfp, green). A, arteries; L, lymphatic vessels; V, veins. (E,E′) Whole-mount limb skin staining of E17.5 Sema3glacZ/+;Nrp2taugfp/+ embryos with antibodies to β-gal (Sema3glacZ, red), GFP (Nrp2taugfp, green), and PECAM1 (blue). Sema3glacZ is expressed in arteries (A, open arrowhead), whereas Nrp2taugfp is expressed in both lymphatic vessels (L, arrow) and veins (V, arrowhead). (F) Schematic model illustrating differential expression patterns of SEMA3s and NRP2 in the skin. Scale bars: 100 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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BIO012302F2: Expression of SEMA3s and their receptor NRP2 in limb skin. (A,B) Expression of Sema3c and Sema3f in E15.5 skin (arrows). In situ hybridization on serial transverse sections was performed with the indicated probes. (C,D) Whole-mount limb skin staining of E17.5 Nrp2taugfp/+ embryos with antibodies to the LEC marker PROX1 (red), LYVE1 (blue), and the EC marker PECAM1 (red) together with GFP (Nrp2taugfp, green). A, arteries; L, lymphatic vessels; V, veins. (E,E′) Whole-mount limb skin staining of E17.5 Sema3glacZ/+;Nrp2taugfp/+ embryos with antibodies to β-gal (Sema3glacZ, red), GFP (Nrp2taugfp, green), and PECAM1 (blue). Sema3glacZ is expressed in arteries (A, open arrowhead), whereas Nrp2taugfp is expressed in both lymphatic vessels (L, arrow) and veins (V, arrowhead). (F) Schematic model illustrating differential expression patterns of SEMA3s and NRP2 in the skin. Scale bars: 100 µm.
Mentions: We next examined the expression patterns of Sema3b, Sema3c, Sema3f and Sema3g in E15.5 skin by in situ hybridization, stages when lymphatic network is actively developing in the skin. We found distinct expression patterns of Sema3c and Sema3f (Fig. 2A,B), albeit a less clear in situ hybridization signal for Sema3b and Sema3g (data not shown). Expression of Sema3c was detectable in the layer adjacent to the boundary between the dermis and hypodermis (Fig. 2A). Consistent with a previous report (Eckhardt and Meyerhans, 1998), expression of Sema3f was observed in the epidermis (Fig. 2B). Although the Sema3g in situ hybridization signal was not clearly detectable, the expression of SEMA3G using Sema3glacZ/+ heterozygous embryos, carrying a lacZ reporter cassette under the endogenous Sema3g promoter, was restricted to arteries and is not expressed in veins or lymphatic vessels within the skin (Fig. 2E,E′; Kutschera et al., 2011).Fig. 2.

Bottom Line: In contrast, Sema3f;Sema3g double mutants display increased lymphatic branching, while Nrp2 mutants exhibit reduced lymphatic branching.Our results provide the first evidence that SEMA3F and SEMA3G function as a negative regulator for dermal lymphangiogenesis in vivo.The reciprocal phenotype in lymphatic branching between Sema3f;Sema3g double mutants and Nrp2 mutants suggest a complex NRP2 function that regulates LEC behavior both positively and negatively, through a binding with VEGFC or SEMA3s.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Stem Cell and Neuro-Vascular Biology, Genetics and Developmental Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Building 10/6C103, 10 Center Drive, Bethesda, MD 20892, USA.

No MeSH data available.


Related in: MedlinePlus