Per2-mediated vascular dysfunction is caused by the upregulation of the connective tissue growth factor (CTGF)

Vaishnavi Jadhav, Qianyi Luo, James M. Dominguez, Jude Al-Sabah, Brahim Chaqour, Maria B. Grant, Ashay Bhatwadekar

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Period 2-mutant mice (Per2M/M), which possess a circadian dysfunction, recapitulate the retinal vascular phenotype similar to diabetic retinopathy (DR). The vascular dysfunction in Per2M/M is associated with an increase in connective tissue growth factor (CTGF/CCN2). At the molecular level, CTGF gene expression is dependent on the canonical Wnt/β-catenin pathway. The nuclear binding of β-catenin to a transcription factor, lymphoid enhancer binding protein (Lef)/T-cell factor (TCF/LEF), leads to downstream activation of CTGF. For this study, we hypothesized that the silencing of Per2 results in nuclear translocation and subsequent transactivation of the CTGF gene. To test this hypothesis, we performed immunofluorescence labeling for CTGF in retinal sections from wild-type (WT) and Per2M/M mice. Human retinal endothelial cells (HRECs) were transfected with siRNA for Per2, and the protein expression of CTGF and β-catenin was evaluated. The TCF/LEF luciferase reporter (TOPflash) assay was performed to validate the involvement of β-catenin in the activation of CTGF. Per2M/M retinas exhibited an increased CTGF immunostaining in ganglion cell layer and retinal endothelium. Silencing of Per2 using siRNA resulted in an upregulation of CTGF and β-catenin. The TOPflash assay revealed an increase in luminescence for HRECs transfected with Per2 siRNA. Our studies show that loss of Per2 results in an activation of CTGF via nuclear entry of β-catenin. Our study provides novel insight into the understanding of microvascular dysfunction in Per2M/M mice.

Original languageEnglish (US)
Article numbere0163367
JournalPLoS One
Volume11
Issue number9
DOIs
StatePublished - Sep 1 2016

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Connective Tissue Growth Factor
blood vessels
connective tissues
growth factors
Blood Vessels
Up-Regulation
Catenins
mice
mutants
small interfering RNA
Small Interfering RNA
Chemical activation
Endothelial cells
endothelial cells
Assays
Endothelial Cells
diabetic retinopathy
TCF Transcription Factors
Retinal Vessels
Wnt Signaling Pathway

ASJC Scopus subject areas

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

Cite this

Per2-mediated vascular dysfunction is caused by the upregulation of the connective tissue growth factor (CTGF). / Jadhav, Vaishnavi; Luo, Qianyi; Dominguez, James M.; Al-Sabah, Jude; Chaqour, Brahim; Grant, Maria B.; Bhatwadekar, Ashay.

In: PLoS One, Vol. 11, No. 9, e0163367, 01.09.2016.

Research output: Contribution to journalArticle

Jadhav, Vaishnavi ; Luo, Qianyi ; Dominguez, James M. ; Al-Sabah, Jude ; Chaqour, Brahim ; Grant, Maria B. ; Bhatwadekar, Ashay. / Per2-mediated vascular dysfunction is caused by the upregulation of the connective tissue growth factor (CTGF). In: PLoS One. 2016 ; Vol. 11, No. 9.
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