Limits...
Neuroinflammation in advanced canine glaucoma.

Jiang B, Harper MM, Kecova H, Adamus G, Kardon RH, Grozdanic SD, Kuehn MH - Mol. Vis. (2010)

Bottom Line: We identified over 500 genes with statistically significant changes in expression level in the glaucomatous retina.Many of the molecules with decreased expression levels have been previously shown to be components of retinal ganglion cells.Our data suggest that in the dog this stage of the disease is accompanied by pronounced retinal neuroinflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52242, USA.

ABSTRACT

Purpose: The pathophysiological events that occur in advanced glaucoma are not well characterized. The principal purpose of this study is to characterize the gene expression changes that occur in advanced glaucoma.

Methods: Retinal RNA was obtained from canine eyes with advanced glaucoma as well as from healthy eyes. Global gene expression patterns were determined using oligonucleotide microarrays and confirmed by real-time PCR. The presence of tumor necrosis factor (TNF) and its receptors was evaluated by immunolabeling. Finally, we evaluated the presence of serum autoantibodies directed against retinal epitopes using western blot analyses.

Results: We identified over 500 genes with statistically significant changes in expression level in the glaucomatous retina. Decreased expression levels were detected for large number of functional groups, including synapse and synaptic transmission, cell adhesion, and calcium metabolism. Many of the molecules with decreased expression levels have been previously shown to be components of retinal ganglion cells. Genes with elevated expression in glaucoma are largely associated with inflammation, such as antigen presentation, protein degradation, and innate immunity. In contrast, expression of many other pro-inflammatory genes, such as interferons or interleukins, was not detected at abnormal levels.

Conclusions: This study characterizes the molecular events that occur in the canine retina with advanced glaucoma. Our data suggest that in the dog this stage of the disease is accompanied by pronounced retinal neuroinflammation.

Show MeSH

Related in: MedlinePlus

Western blotting analysis of autoantibodies present in sera of dogs with glaucoma and in sera of control healthy dog sera. Sera from dogs with glaucoma are labeled from 1 to 7; control dog sera are labeled as “control dog sera,” Lanes R- recoverin, E – enolase, C - crystalline-μ are immunostained with antibodies specifically directed against these molecules. N – negative control, M – molecular weight markers.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2965571&req=5

f7: Western blotting analysis of autoantibodies present in sera of dogs with glaucoma and in sera of control healthy dog sera. Sera from dogs with glaucoma are labeled from 1 to 7; control dog sera are labeled as “control dog sera,” Lanes R- recoverin, E – enolase, C - crystalline-μ are immunostained with antibodies specifically directed against these molecules. N – negative control, M – molecular weight markers.

Mentions: Despite the apparent paucity of CD3/CD79 positive cells, it was conceivable that canine glaucoma may result in the formation of autoantibodies directed against retinal antigens. To determine if this does indeed occur, we incubated western blots of retinal protein extracts with serum obtained from seven dogs with moderate to advanced glaucoma and ten healthy control animals (Figure 7). While minor immune reactivity can be observed in all samples, the majority of the serum obtained from dogs with glaucoma reacts with retinal proteins more vigorously than that obtained from healthy control dogs. It is noteworthy that the samples with the most pronounced immunoreactivity were derived from dogs with advanced glaucoma. A general pattern of labeled bands is not apparent, rather it appears that each individual displays immunereactivity to a specific subset of molecules. In the majority of cases the labeled molecules do not react with purified Recoverin, α-enolase, or crystallins, suggesting that these proteins are not major autoantigens in dogs with glaucoma.


Neuroinflammation in advanced canine glaucoma.

Jiang B, Harper MM, Kecova H, Adamus G, Kardon RH, Grozdanic SD, Kuehn MH - Mol. Vis. (2010)

Western blotting analysis of autoantibodies present in sera of dogs with glaucoma and in sera of control healthy dog sera. Sera from dogs with glaucoma are labeled from 1 to 7; control dog sera are labeled as “control dog sera,” Lanes R- recoverin, E – enolase, C - crystalline-μ are immunostained with antibodies specifically directed against these molecules. N – negative control, M – molecular weight markers.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2965571&req=5

f7: Western blotting analysis of autoantibodies present in sera of dogs with glaucoma and in sera of control healthy dog sera. Sera from dogs with glaucoma are labeled from 1 to 7; control dog sera are labeled as “control dog sera,” Lanes R- recoverin, E – enolase, C - crystalline-μ are immunostained with antibodies specifically directed against these molecules. N – negative control, M – molecular weight markers.
Mentions: Despite the apparent paucity of CD3/CD79 positive cells, it was conceivable that canine glaucoma may result in the formation of autoantibodies directed against retinal antigens. To determine if this does indeed occur, we incubated western blots of retinal protein extracts with serum obtained from seven dogs with moderate to advanced glaucoma and ten healthy control animals (Figure 7). While minor immune reactivity can be observed in all samples, the majority of the serum obtained from dogs with glaucoma reacts with retinal proteins more vigorously than that obtained from healthy control dogs. It is noteworthy that the samples with the most pronounced immunoreactivity were derived from dogs with advanced glaucoma. A general pattern of labeled bands is not apparent, rather it appears that each individual displays immunereactivity to a specific subset of molecules. In the majority of cases the labeled molecules do not react with purified Recoverin, α-enolase, or crystallins, suggesting that these proteins are not major autoantigens in dogs with glaucoma.

Bottom Line: We identified over 500 genes with statistically significant changes in expression level in the glaucomatous retina.Many of the molecules with decreased expression levels have been previously shown to be components of retinal ganglion cells.Our data suggest that in the dog this stage of the disease is accompanied by pronounced retinal neuroinflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52242, USA.

ABSTRACT

Purpose: The pathophysiological events that occur in advanced glaucoma are not well characterized. The principal purpose of this study is to characterize the gene expression changes that occur in advanced glaucoma.

Methods: Retinal RNA was obtained from canine eyes with advanced glaucoma as well as from healthy eyes. Global gene expression patterns were determined using oligonucleotide microarrays and confirmed by real-time PCR. The presence of tumor necrosis factor (TNF) and its receptors was evaluated by immunolabeling. Finally, we evaluated the presence of serum autoantibodies directed against retinal epitopes using western blot analyses.

Results: We identified over 500 genes with statistically significant changes in expression level in the glaucomatous retina. Decreased expression levels were detected for large number of functional groups, including synapse and synaptic transmission, cell adhesion, and calcium metabolism. Many of the molecules with decreased expression levels have been previously shown to be components of retinal ganglion cells. Genes with elevated expression in glaucoma are largely associated with inflammation, such as antigen presentation, protein degradation, and innate immunity. In contrast, expression of many other pro-inflammatory genes, such as interferons or interleukins, was not detected at abnormal levels.

Conclusions: This study characterizes the molecular events that occur in the canine retina with advanced glaucoma. Our data suggest that in the dog this stage of the disease is accompanied by pronounced retinal neuroinflammation.

Show MeSH
Related in: MedlinePlus