May 2006
Volume 47, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2006
Transcriptional Alterations in Müller Cells With Elevated Glucose
Author Affiliations & Notes
  • T.W. Gardner
    Penn State College of Medicine, Hershey, PA
    Department of Ophthalmology,
    Department of Cellular & Molecular Physiology,
  • D.J. Krissinger
    Penn State College of Medicine, Hershey, PA
    Functional Genomics Core Facility,
  • T.L. Schrufer
    Penn State College of Medicine, Hershey, PA
    Department of Cellular & Molecular Physiology,
  • R.M. Brucklacher
    Penn State College of Medicine, Hershey, PA
    Functional Genomics Core Facility,
  • S.R. Kimball
    Penn State College of Medicine, Hershey, PA
    Department of Cellular & Molecular Physiology,
  • W.M. Freeman
    Penn State College of Medicine, Hershey, PA
    Department of Pharmacology,
  • Footnotes
    Commercial Relationships  T.W. Gardner, None; D.J. Krissinger, None; T.L. Schrufer, None; R.M. Brucklacher, None; S.R. Kimball, None; W.M. Freeman, None.
  • Footnotes
    Support  JDRF Penn State Diabetic Retinopathy Center and JDRF Program Project Grant
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 1722. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      T.W. Gardner, D.J. Krissinger, T.L. Schrufer, R.M. Brucklacher, S.R. Kimball, W.M. Freeman; Transcriptional Alterations in Müller Cells With Elevated Glucose . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1722.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: : Müller cells play a critical role in the retina by serving as the primary glial cell. Neurodegeneration is a pathological feature of diabetic retinopathy. One key element of neurodegeneration is a reduction in the number of synapses between neurons. The aim of this study was to test the hypothesis that diabetes alters the expression of specific synaptic and structural proteins in the rat retina.

Methods: : The Müller cell line TR–MUL5 was maintained in culture medium containing 5 mM glucose. On the day of the study, cells were incubated in medium containing either 25mM glucose or mannitol for 3 or 12 hours and RNA was isolated from the cells. RNA was subjected to whole genome microarray analysis using Codelink arrays. Statistical and ontological analysis was performed on the array data to determine processes and pathways affected by elevated glucose. Genes identified as regulated by microarray analysis were then confirmed by quantitative PCR using TaqMan fluorescent probes and real–time detection.

Results: : Of the 33911 probes on the array, 21168 were present at a detectable level. 78 genes were found to be significantly different between glucose and mannitol conditions at 3 hours and 186 were found to be altered at 12 hours. Of these changes a significant increased abundance of vesicle and cytoskeletal genes were found at the 12 hour timepoint. Quantitative PCR analysis confirmed significantly higher expression of the NGFI–A, NGFI–B, Fos, and VEGFA mRNAs, and significantly lower expression of Igfbp2. In addition, elevated expression of Jundp2 and reduced expression of KIFC2 were confirmed at 12 hours.

Conclusions: : Elevated concentrations of glucose cause a number of significant changes in gene expression in cultures of Müller cells within hours of treatment. Further experiments must be performed to examine these changes in Müller cells within the retina.

Keywords: diabetic retinopathy • Muller cells • gene microarray 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×