title: The Zeb proteins δEF1 and Sip1 may have distinct functions in lens cells following cataract surgery.
abstract: OBJECTIVE
  Posterior capsular opacification (PCO), the most prevalent side effect of cataract surgery, occurs when residual lens epithelial cells (LECs) undergo fiber cell differentiation or epithelial-to-mesenchymal transition (EMT). Here, we used a murine cataract surgery model to investigate the role of the Zeb proteins, Smad interacting protein 1 (Sip1) and δ-crystallin enhancer-binding factor 1 (δEF1), during PCO.
  
  
  METHODS
  Extracapsular extraction of lens fiber cells was performed on wild-type and Sip1 knockout mice. Protein expression patterns were assessed at multiple time points after surgery using confocal immunofluorescence. βB1-Crystallin mRNA levels were measured using quantitative RT-PCR. We used Transfac searches to identify δEF1 binding sites in the βB1-crystallin promoter and transfection analysis to test the ability of δEF1 to regulate βB1-crystallin expression.
  
  
  RESULTS
  δEF1, which, in other systems, can activate fibrotic genes (e.g., α-smooth muscle actin) and repress epithelial genes, upregulates by 48 hours after fiber cell removal. In culture, δEF1 repressed βB1-crystallin promoter activity, suggesting that it may also turn off lens gene expression following surgery, contributing to "fibrotic PCO" development. Sip1 also upregulates in LECs by 48 hours, but analysis of Sip1 knockout lenses demonstrated that Sip1 does not play a major role in EMT or fiber cell differentiation after surgery. However, Sip1 knockout LECs do express the ectodermal marker keratin 8, suggesting that Sip1 may limit the reprogramming of residual LECs to an embryonic state.
  
  
  CONCLUSIONS
  Zeb transcription factors likely play important, but distinct roles in PCO development after cataract surgery.
authors.affiliation: Department of Biological Sciences, University of Delaware, Newark, Delaware, United States.
authors.forename: Abby L
authors.initials: AL
authors.lastname: Manthey
authors.affiliation: Department of Biological Sciences, University of Delaware, Newark, Delaware, United States.
authors.forename: Anne M
authors.initials: AM
authors.lastname: Terrell
authors.affiliation: Department of Biological Sciences, University of Delaware, Newark, Delaware, United States.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Biological Sciences, University of Delaware, Newark, Delaware, United States.
authors.forename: Jennifer R
authors.initials: JR
authors.lastname: Taube
authors.affiliation: Department of Biological Sciences, University of Delaware, Newark, Delaware, United States.
authors.forename: Alisha R
authors.initials: AR
authors.lastname: Yallowitz
authors.affiliation: Department of Biological Sciences, University of Delaware, Newark, Delaware, United States.
authors.forename: Melinda K
authors.initials: MK
authors.lastname: Duncan
mesh_terms: D000818:Animals; D002386:Cataract / Q000378:metabolism* / Q000473:pathology; D002387:Cataract Extraction / Q000009:adverse effects*; D004195:Disease Models, Animal; D018398:Homeodomain Proteins / Q000502:physiology*; D007908:Lens, Crystalline / Q000378:metabolism*; D018345:Mice, Knockout; D009419:Nerve Tissue Proteins / Q000502:physiology*; D011183:Postoperative Complications; D012333:RNA, Messenger / Q000378:metabolism; D014157:Transcription Factors / Q000502:physiology*; D000071799:Zinc Finger E-box-Binding Homeobox 1
chemical_list: D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; C491561:Sip1 protein, mouse; C519422:ZEB1 protein, mouse; D000071799:Zinc Finger E-box-Binding Homeobox 1
keywords: Zeb; cataract surgery; epithelial-to-mesenchymal transition; lens regeneration; mouse disease model; posterior capsular opacification
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