TY - JOUR
T1 - Molecular profiling of the developing lacrimal gland reveals putative role of notch signaling in branching morphogenesis
AU - Dvoriantchikova, Galina
AU - Tao, Wensi
AU - Pappas, Steve
AU - Gaidosh, Gabriel
AU - Tse, David T.
AU - Ivanov, Dmitry
AU - Pelaez, Daniel
N1 - Funding Information:
The authors thank the Analytic Imaging Facility at the Bascom Palmer Eye Institute (BPEI). This study was possible due to the generous and visionary support of Nasser Ibrahim Al-Rashid, PhD. Supported by grants in part from Nasser Al-Rashid Orbital Vision Research Center Grant (DP; Miami, FL, USA), GemCon Family Foundation Research Grant (DP; Palm Beach, FL, USA), National Eye Institute/National Institutes of Health (NIH) Grant R01 EY022348 (DI), and NIH Center Core Grant P30EY014801 (Bethesda, MD, USA).
Publisher Copyright:
© 2017 The Authors.
Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
PY - 2017/2/1
Y1 - 2017/2/1
N2 - PURPOSE. Although normal function of the lacrimal gland is essential for vision (and thus for human well-being), the lacrimal gland remains rather poorly understood at a molecular level. The purpose of this study was to identify new genes and signaling cascades involved in lacrimal gland development. METHODS. To identify these genes, we used microarray analysis to compare the gene expression profiles of developing (embryonic) and adult lacrimal glands. Differential data were validated by quantitative RT-PCR, and several corresponding proteins were confirmed by immunohistochemistry and Western blot analysis. To evaluate the role of NOTCH signaling in lacrimal gland (LG) development, we used the NOTCH inhibitor DAPT and conditional Notch1 knockouts. RESULTS. Our microarray data and an in silico reconstruction of cellular networks revealed significant changes in the expression patterns of genes from the NOTCH, WNT, TGFb, and Hedgehog pathways, all of which are involved in the regulation of epithelial-to-mesenchymal transition (EMT). Our study also revealed new putative lacrimal gland stem cell/progenitor markers. We found that inhibiting Notch signaling both increases the average number of lacrimal gland lobules and reduces the size of each lobule. CONCLUSIONS. Our findings suggest that NOTCH-, WNT-, TGFb-, and Hedgehog-regulated EMT transition are critical mechanisms in lacrimal gland development and morphogenesis. Our data also supports the hypothesis that NOTCH signaling regulates branching morphogenesis in the developing lacrimal gland by suppressing cleft formation.
AB - PURPOSE. Although normal function of the lacrimal gland is essential for vision (and thus for human well-being), the lacrimal gland remains rather poorly understood at a molecular level. The purpose of this study was to identify new genes and signaling cascades involved in lacrimal gland development. METHODS. To identify these genes, we used microarray analysis to compare the gene expression profiles of developing (embryonic) and adult lacrimal glands. Differential data were validated by quantitative RT-PCR, and several corresponding proteins were confirmed by immunohistochemistry and Western blot analysis. To evaluate the role of NOTCH signaling in lacrimal gland (LG) development, we used the NOTCH inhibitor DAPT and conditional Notch1 knockouts. RESULTS. Our microarray data and an in silico reconstruction of cellular networks revealed significant changes in the expression patterns of genes from the NOTCH, WNT, TGFb, and Hedgehog pathways, all of which are involved in the regulation of epithelial-to-mesenchymal transition (EMT). Our study also revealed new putative lacrimal gland stem cell/progenitor markers. We found that inhibiting Notch signaling both increases the average number of lacrimal gland lobules and reduces the size of each lobule. CONCLUSIONS. Our findings suggest that NOTCH-, WNT-, TGFb-, and Hedgehog-regulated EMT transition are critical mechanisms in lacrimal gland development and morphogenesis. Our data also supports the hypothesis that NOTCH signaling regulates branching morphogenesis in the developing lacrimal gland by suppressing cleft formation.
KW - Branching morphogenesis
KW - Development
KW - Lacrimal gland
KW - Microarray analysis
KW - NOTCH signaling
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U2 - 10.1167/iovs.16-20315
DO - 10.1167/iovs.16-20315
M3 - Article
C2 - 28192800
AN - SCOPUS:85012260176
VL - 58
SP - 1098
EP - 1109
JO - Investigative Ophthalmology and Visual Science
JF - Investigative Ophthalmology and Visual Science
SN - 0146-0404
IS - 2
ER -