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(Investigative Ophthalmology and Visual Science. 2006;47:3860-3869.)
© 2006 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.05-1408
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Pressure-Induced Regulation of IL-6 in Retinal Glial Cells: Involvement of the Ubiquitin/Proteasome Pathway and NF{kappa}B

Rebecca M. Sappington and David J. Calkins

From the Department of Ophthalmology and Visual Sciences, The Vanderbilt Eye Institute, Vanderbilt University Medical Center, Nashville, Tennessee.

PURPOSE. To investigate how hydrostatic pressure influences regulation of interleukin (IL)-6 by retinal glia and whether this regulation is associated with the ubiquitin/proteasome pathway (UPP) and activation of the transcription factor nuclear factor (NF){kappa}B.

METHODS. Astrocytes and microglia isolated from rat retina were maintained in vitro, and the IL-6 concentration in the media at ambient and elevated pressure were compared, with and without the proteasome inhibitor MG132 (10 µM). Immunocytochemistry was used to correlate translocation of NF{kappa}B with pressure.

RESULTS. Exposure to elevated pressure for 24 hours maximally altered the concentration of media IL-6 of glia cultures, where IL-6 concentrations decreased in astrocyte cultures and increased in microglia cultures. These pressure-induced changes in IL-6 were largely insensitive to MG132 in astrocytes, but were largely MG132-sensitive in microglia. Like IL-6 regulation, pressure-induced activation of NF{kappa}B also differed between the two glial cell types, where nuclear localization of NF{kappa}B was transient in astrocytes, but sustained in microglia. Elevated pressure also increased MG132-sensitive expression of IL-6 mRNA by microglia.

CONCLUSIONS. Though pressure-induced regulation of IL-6 by astrocytes is preceded by NF{kappa}B translocation, it is not altered by MG132 and therefore is not likely to be regulated by NF{kappa}B or the UPP. In contrast, pressure-induced regulation of IL-6 protein and mRNA by microglia is preceded by NF{kappa}B translocation and is sensitive to MG132. Together with precedence in the literature, these data suggest that pressure-induced activation of the UPP leads to transcription of IL-6 driven by NF{kappa}B.




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R. M. Sappington and D. J. Calkins
Contribution of TRPV1 to Microglia-Derived IL-6 and NF{kappa}B Translocation with Elevated Hydrostatic Pressure
Invest. Ophthalmol. Vis. Sci., July 1, 2008; 49(7): 3004 - 3017.
[Abstract] [Full Text] [PDF]


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