Attribution 4.0 International (CC BY 4.0)2022-09-302022-09-302017-09-22Wang, J., Ma, H. Y., Krishnamoorthy, R. R., Yorio, T., & He, S. (2017). A feed-forward regulation of endothelin receptors by c-Jun in human non-pigmented ciliary epithelial cells and retinal ganglion cells. PloS one, 12(9), e0185390. https://doi.org/10.1371/journal.pone.01853901932-6203https://hdl.handle.net/20.500.12503/31815c-Jun, c-Jun N-terminal kinase(JNK) and endothelin B (ETB) receptor have been shown to contribute to the pathogenesis of glaucoma. Previously, we reported that an increase of c-Jun and CCAAT/enhancer binding protein beta (C/EBPbeta) immunohistostaining is associated with upregulation of the ETB receptor within the ganglion cell layer of rats with elevated intraocular pressure (IOP). In addition, both transcription factors regulate the expression of the ETB receptor in human non-pigmented ciliary epithelial cells (HNPE). The current study addressed the mechanisms by which ET-1 produced upregulation of ET receptors in primary rat retinal ganglion cells (RGCs) and HNPE cells. Treatment of ET-1 and ET-3 increased the immunocytochemical staining of c-Jun and C/EBPbeta in primary rat RGCs and co-localization of both transcription factors was observed. A marked increase in DNA binding activity of AP-1 and C/EBPbeta as well as elevated protein levels of c-Jun and c-Jun-N-terminal kinase (JNK) were detected following ET-1 treatment in HNPE cells. Overexpression of ETA or ETB receptor promoted the upregulation of c-Jun and also elevated its promoter activity. In addition, upregulation of C/EBPbeta augmented DNA binding and mRNA expression of c-Jun, and furthermore, the interaction of c-Jun and C/EBPbeta was confirmed using co-immunoprecipitation. Apoptosis of HNPE cells was identified following ET-1 treatment, and overexpression of the ETA or ETB receptor produced enhanced apoptosis. ET-1 mediated upregulation of c-Jun and C/EBPbeta and their interaction may represent a novel mechanism contributing to the regulation of endothelin receptor expression. Reciprocally, c-Jun was also found to regulate the ET receptors and C/EBPbeta appeared to play a regulatory role in promoting expression of c-Jun. Taken together, the data suggests that ET-1 triggers the upregulation of c-Jun through both ETA and ETB receptors, and conversely c-Jun also upregulates endothelin receptor expression, thereby generating a positive feed-forward loop of endothelin receptor activation and expression. This feed-forward regulation may contribute to RGC death and astrocyte proliferation following ET-1 treatment.http://creativecommons.org/licenses/by/4.0/AnimalsApoptosis / physiologyCCAAT-Enhancer-Binding Protein-beta / metabolismCells, CulturedCilia / enzymologyEndothelin-1 / metabolismEpithelial Cells / enzymologyHumansJNK Mitogen-Activated Protein Kinases / metabolismProtein BindingRats, Sprague-DawleyReceptors, Endothelin / metabolismRetinal Ganglion Cells / enzymologyTranscription Factor AP-1 / metabolismArticle© 2017 Wang et al.129