Expression of acute-phase response proteins in retinal Müller cells in diabetes.
OBJECTIVE
To characterize the whole spectrum of gene expression changes induced by diabetes in retinal Müller glial cells.


METHODS
Müller cells were isolated from the retina of streptozotocin-diabetic and age-matched control rats by gradient centrifugation and immediately processed for RNA isolation. The gene expression profile of Müller cells was studied with the GeneChip Rat Genome oligonucleotide array (Affymetrix, Santa Clara, CA). The upregulation of acute-phase proteins in the retina of diabetic rats was confirmed by Northern and Western blot analyses. Real-time-RT-PCR was used to study the retinal expression of inflammatory cytokines.


RESULTS
Gene expression profiling identified 78 genes as differentially expressed in diabetic Müller cells. One third of these genes were associated with inflammation, including a large cluster (18% of the differentially expressed genes) of acute-phase response proteins: alpha2-macroglobulin, ceruloplasmin, complement components, lipocalin-2, metallothionein, serine protease inhibitor-2, transferrin, tissue inhibitor of metalloproteases-1, transthyretin, and the transcription factor C/EBPdelta. Northern and Western blot analyses confirmed the upregulation of alpha2-macroglobulin and ceruloplasmin in the diabetic retina, but not in the cerebral cortex and liver of the same animals. The acute-phase response of Müller cells in diabetes was associated with upregulation of interleukin (IL)-1beta in the retina.


CONCLUSIONS
Müller cells acquire a complex and specific reactive phenotype in diabetes characterized by the induction of acute-phase response proteins and other inflammation-related genes. The concomitant upregulation of IL-1beta in the retina of diabetic rats points to this cytokine as a possible mediator of the acute-phase response mounted by Müller cells in diabetes.
Gerhardinger
Chiara
C
Schepens Eye Research Institute, Harvard Medical School, Boston, MA 02114, USA. chiara@vision.eri.harvard.edu
Costa
Montserrat Biarnés
MB
Coulombe
Mary C
MC
Toth
Ildiko
I
Hoehn
Todd
T
Grosu
Paul
P
D000209:Acute-Phase Proteins / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism*; D003930:Diabetic Retinopathy / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000378:metabolism; D008297:Male; D009457:Neuroglia / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000473:pathology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
D000209:Acute-Phase Proteins; D007375:Interleukin-1; D012333:RNA, Messenger; D012313:RNA