Activation of Dll4/notch signaling and hypoxia-inducible factor-1 alpha facilitates lymphangiogenesis in lacrimal glands in dry eye

Ji Hwan Min, Chul Hee Lee, Yong Woo Ji, Areum Yeo, Hyemi Noh, Insil Song, Eung Kweon Kim, Hyung Keun Lee

Research output: Contribution to journalArticle

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Abstract

Purpose By using hypoxia-inducible factor-1 alpha conditional knockout (HIF-1α CKO) mice and a dry eye (DE) mouse model, we aimed to determine the role played by delta-like ligand 4 (Dll4)/Notch signaling and HIF-1α in the lymphangiogenesis of lacrimal glands (LGs). Methods C57BL/6 mice were housed in a controlled-environment chamber for DE induction. During DE induction, the expression level of Dll4/Notch signaling and lymphangiogenesis in LGs was measured by quantitative RT-PCR, immunoblot, and immunofluorescence staining. Next, lymphangiogenesis was measured after Dll4/Notch signal inhibition by anti-Dll4 antibody or I-secretase inhibitor. Using HIF-1α CKO mice, the expression of Dll4/Notch signaling and lymphangiogenesis in LGs of DE-induced HIF-1α CKO mice were assessed. Additionally, the infiltration of CD45+ cells in LGs was assessed by immunohistochemical (IHC) staining and flow cytometry for each condition. Results DE significantly upregulated Dll4/Notch and lymphangiogenesis in LGs. Inhibition of Dll4/ Notch significantly suppressed lymphangiogenesis in LGs. Compared to wild-type (WT) mice, DE induced HIF-1α CKO mice showed markedly low levels of Dll4/Notch and lymphangiogenesis. Inhibition of lymphangiogenesis by Dll4/Notch suppression resulted in increased CD45+ cell infiltration in LGs. Likewise, CD45+ cells infiltrated more in the LGs of HIF-1α CKO DE mice than in non-DE HIF-1α CKO mice. Conclusions Dll4/Notch signaling and HIF-1α are closely related to lymphangiogenesis in DE-induced LGs. Lymphangiogenesis stimulated by Dll4/Notch and HIF-1α may play a role in protecting LGs from DE-induced inflammation by aiding the clearance of immune cells from LGs.

Original languageEnglish
Article numbere0147846
JournalPloS one
Volume11
Issue number2
DOIs
Publication statusPublished - 2016 Feb 1

Fingerprint

Lymphangiogenesis
lacrimal apparatus
Hypoxia-Inducible Factor 1
Lacrimal Apparatus
eyes
Chemical activation
Knockout Mice
mice
Infiltration
delta protein
ligands
hypoxia-inducible factor 1
cells
Staining and Labeling
Controlled Environment
Amyloid Precursor Protein Secretases
Flow cytometry
Inbred C57BL Mouse
fluorescent antibody technique
Fluorescent Antibody Technique

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Min, Ji Hwan ; Lee, Chul Hee ; Ji, Yong Woo ; Yeo, Areum ; Noh, Hyemi ; Song, Insil ; Kim, Eung Kweon ; Lee, Hyung Keun. / Activation of Dll4/notch signaling and hypoxia-inducible factor-1 alpha facilitates lymphangiogenesis in lacrimal glands in dry eye. In: PloS one. 2016 ; Vol. 11, No. 2.
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title = "Activation of Dll4/notch signaling and hypoxia-inducible factor-1 alpha facilitates lymphangiogenesis in lacrimal glands in dry eye",
abstract = "Purpose By using hypoxia-inducible factor-1 alpha conditional knockout (HIF-1{\^I}± CKO) mice and a dry eye (DE) mouse model, we aimed to determine the role played by delta-like ligand 4 (Dll4)/Notch signaling and HIF-1{\^I}± in the lymphangiogenesis of lacrimal glands (LGs). Methods C57BL/6 mice were housed in a controlled-environment chamber for DE induction. During DE induction, the expression level of Dll4/Notch signaling and lymphangiogenesis in LGs was measured by quantitative RT-PCR, immunoblot, and immunofluorescence staining. Next, lymphangiogenesis was measured after Dll4/Notch signal inhibition by anti-Dll4 antibody or I-secretase inhibitor. Using HIF-1{\^I}± CKO mice, the expression of Dll4/Notch signaling and lymphangiogenesis in LGs of DE-induced HIF-1{\^I}± CKO mice were assessed. Additionally, the infiltration of CD45+ cells in LGs was assessed by immunohistochemical (IHC) staining and flow cytometry for each condition. Results DE significantly upregulated Dll4/Notch and lymphangiogenesis in LGs. Inhibition of Dll4/ Notch significantly suppressed lymphangiogenesis in LGs. Compared to wild-type (WT) mice, DE induced HIF-1{\^I}± CKO mice showed markedly low levels of Dll4/Notch and lymphangiogenesis. Inhibition of lymphangiogenesis by Dll4/Notch suppression resulted in increased CD45+ cell infiltration in LGs. Likewise, CD45+ cells infiltrated more in the LGs of HIF-1{\^I}± CKO DE mice than in non-DE HIF-1{\^I}± CKO mice. Conclusions Dll4/Notch signaling and HIF-1{\^I}± are closely related to lymphangiogenesis in DE-induced LGs. Lymphangiogenesis stimulated by Dll4/Notch and HIF-1{\^I}± may play a role in protecting LGs from DE-induced inflammation by aiding the clearance of immune cells from LGs.",
author = "Min, {Ji Hwan} and Lee, {Chul Hee} and Ji, {Yong Woo} and Areum Yeo and Hyemi Noh and Insil Song and Kim, {Eung Kweon} and Lee, {Hyung Keun}",
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Activation of Dll4/notch signaling and hypoxia-inducible factor-1 alpha facilitates lymphangiogenesis in lacrimal glands in dry eye. / Min, Ji Hwan; Lee, Chul Hee; Ji, Yong Woo; Yeo, Areum; Noh, Hyemi; Song, Insil; Kim, Eung Kweon; Lee, Hyung Keun.

In: PloS one, Vol. 11, No. 2, e0147846, 01.02.2016.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Activation of Dll4/notch signaling and hypoxia-inducible factor-1 alpha facilitates lymphangiogenesis in lacrimal glands in dry eye

AU - Min, Ji Hwan

AU - Lee, Chul Hee

AU - Ji, Yong Woo

AU - Yeo, Areum

AU - Noh, Hyemi

AU - Song, Insil

AU - Kim, Eung Kweon

AU - Lee, Hyung Keun

PY - 2016/2/1

Y1 - 2016/2/1

N2 - Purpose By using hypoxia-inducible factor-1 alpha conditional knockout (HIF-1α CKO) mice and a dry eye (DE) mouse model, we aimed to determine the role played by delta-like ligand 4 (Dll4)/Notch signaling and HIF-1α in the lymphangiogenesis of lacrimal glands (LGs). Methods C57BL/6 mice were housed in a controlled-environment chamber for DE induction. During DE induction, the expression level of Dll4/Notch signaling and lymphangiogenesis in LGs was measured by quantitative RT-PCR, immunoblot, and immunofluorescence staining. Next, lymphangiogenesis was measured after Dll4/Notch signal inhibition by anti-Dll4 antibody or I-secretase inhibitor. Using HIF-1α CKO mice, the expression of Dll4/Notch signaling and lymphangiogenesis in LGs of DE-induced HIF-1α CKO mice were assessed. Additionally, the infiltration of CD45+ cells in LGs was assessed by immunohistochemical (IHC) staining and flow cytometry for each condition. Results DE significantly upregulated Dll4/Notch and lymphangiogenesis in LGs. Inhibition of Dll4/ Notch significantly suppressed lymphangiogenesis in LGs. Compared to wild-type (WT) mice, DE induced HIF-1α CKO mice showed markedly low levels of Dll4/Notch and lymphangiogenesis. Inhibition of lymphangiogenesis by Dll4/Notch suppression resulted in increased CD45+ cell infiltration in LGs. Likewise, CD45+ cells infiltrated more in the LGs of HIF-1α CKO DE mice than in non-DE HIF-1α CKO mice. Conclusions Dll4/Notch signaling and HIF-1α are closely related to lymphangiogenesis in DE-induced LGs. Lymphangiogenesis stimulated by Dll4/Notch and HIF-1α may play a role in protecting LGs from DE-induced inflammation by aiding the clearance of immune cells from LGs.

AB - Purpose By using hypoxia-inducible factor-1 alpha conditional knockout (HIF-1α CKO) mice and a dry eye (DE) mouse model, we aimed to determine the role played by delta-like ligand 4 (Dll4)/Notch signaling and HIF-1α in the lymphangiogenesis of lacrimal glands (LGs). Methods C57BL/6 mice were housed in a controlled-environment chamber for DE induction. During DE induction, the expression level of Dll4/Notch signaling and lymphangiogenesis in LGs was measured by quantitative RT-PCR, immunoblot, and immunofluorescence staining. Next, lymphangiogenesis was measured after Dll4/Notch signal inhibition by anti-Dll4 antibody or I-secretase inhibitor. Using HIF-1α CKO mice, the expression of Dll4/Notch signaling and lymphangiogenesis in LGs of DE-induced HIF-1α CKO mice were assessed. Additionally, the infiltration of CD45+ cells in LGs was assessed by immunohistochemical (IHC) staining and flow cytometry for each condition. Results DE significantly upregulated Dll4/Notch and lymphangiogenesis in LGs. Inhibition of Dll4/ Notch significantly suppressed lymphangiogenesis in LGs. Compared to wild-type (WT) mice, DE induced HIF-1α CKO mice showed markedly low levels of Dll4/Notch and lymphangiogenesis. Inhibition of lymphangiogenesis by Dll4/Notch suppression resulted in increased CD45+ cell infiltration in LGs. Likewise, CD45+ cells infiltrated more in the LGs of HIF-1α CKO DE mice than in non-DE HIF-1α CKO mice. Conclusions Dll4/Notch signaling and HIF-1α are closely related to lymphangiogenesis in DE-induced LGs. Lymphangiogenesis stimulated by Dll4/Notch and HIF-1α may play a role in protecting LGs from DE-induced inflammation by aiding the clearance of immune cells from LGs.

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