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Gene expression profiling of vasoregression in the retina--involvement of microglial cells.

Feng Y, Wang Y, Li L, Wu L, Hoffmann S, Gretz N, Hammes HP - PLoS ONE (2011)

Bottom Line: Of the 157 genes regulated more than twofold, the MHC class II invariant chain CD74 yielded the strongest upregulation, and was allocated to activated microglial cells close to the vessels undergoing vasoregression.Pathway clustering identified genes of the immune system including inflammatory signaling, and components of the complement cascade upregulated during vasoregression.Together, our data suggest that microglial cells involved in retinal immune response participate in the initiation of vasoregression in the retina.

View Article: PubMed Central - PubMed

Affiliation: 5 Medical Department, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany.

ABSTRACT
Vasoregression is a hallmark of vascular eye diseases but the mechanisms involved are still largely unknown. We have recently characterized a rat ciliopathy model which develops primary photoreceptor degeneration and secondary vasoregression. To improve the understanding of secondary vasoregression in retinal neurodegeneration, we used microarray techniques to compare gene expression profiles in this new model before and after retinal vasoregression. Differential gene expression was validated by quantitative RT-PCR, Western blot and immunofluorescence. Of the 157 genes regulated more than twofold, the MHC class II invariant chain CD74 yielded the strongest upregulation, and was allocated to activated microglial cells close to the vessels undergoing vasoregression. Pathway clustering identified genes of the immune system including inflammatory signaling, and components of the complement cascade upregulated during vasoregression. Together, our data suggest that microglial cells involved in retinal immune response participate in the initiation of vasoregression in the retina.

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Expression of CD74 in SD and TGR retinas at 1- and 3 months.A. Translational analysis of CD74 by Western blot. B. quantitative analysis of translational CD74 expression. The values at 1-month are standardized to 1. C: expression and localization of CD74 in the retina. Vessels are visualized by staining with Lectin (red), and CD74 positive cells are displayed in green. Few CD74 positive cells are seen in SD in the superficial and deep capillary layers. A slight increase in number of CD74 positive cells is found in TGR in the superficial capillary layer at 1 month. The numbers of CD74 positive cells are raised in the deep capillary layer in 3-month TGR compared with other three groups at all time points observed. C–F: the superficial capillary layer; G–J: the deep capillary layer. 1 m: 1-month retina. 3 m: 3-month retina. *p<0.05: 3-month TGR compared with other three groups.
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pone-0016865-g005: Expression of CD74 in SD and TGR retinas at 1- and 3 months.A. Translational analysis of CD74 by Western blot. B. quantitative analysis of translational CD74 expression. The values at 1-month are standardized to 1. C: expression and localization of CD74 in the retina. Vessels are visualized by staining with Lectin (red), and CD74 positive cells are displayed in green. Few CD74 positive cells are seen in SD in the superficial and deep capillary layers. A slight increase in number of CD74 positive cells is found in TGR in the superficial capillary layer at 1 month. The numbers of CD74 positive cells are raised in the deep capillary layer in 3-month TGR compared with other three groups at all time points observed. C–F: the superficial capillary layer; G–J: the deep capillary layer. 1 m: 1-month retina. 3 m: 3-month retina. *p<0.05: 3-month TGR compared with other three groups.

Mentions: To investigate translational expression levels of CD74 obtained on the microarray, we performed western blot analysis with retinal lysates from TGR and SD rats at 1 and 3 months. Weak CD74 expression was found in the retinas of 1- and 3-month SD and 1-month TGR rats. A significant overexpression of CD74 protein was detected in 3-month TGR retinas compared with other three groups (Figure 5A and B).


Gene expression profiling of vasoregression in the retina--involvement of microglial cells.

Feng Y, Wang Y, Li L, Wu L, Hoffmann S, Gretz N, Hammes HP - PLoS ONE (2011)

Expression of CD74 in SD and TGR retinas at 1- and 3 months.A. Translational analysis of CD74 by Western blot. B. quantitative analysis of translational CD74 expression. The values at 1-month are standardized to 1. C: expression and localization of CD74 in the retina. Vessels are visualized by staining with Lectin (red), and CD74 positive cells are displayed in green. Few CD74 positive cells are seen in SD in the superficial and deep capillary layers. A slight increase in number of CD74 positive cells is found in TGR in the superficial capillary layer at 1 month. The numbers of CD74 positive cells are raised in the deep capillary layer in 3-month TGR compared with other three groups at all time points observed. C–F: the superficial capillary layer; G–J: the deep capillary layer. 1 m: 1-month retina. 3 m: 3-month retina. *p<0.05: 3-month TGR compared with other three groups.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3040753&req=5

pone-0016865-g005: Expression of CD74 in SD and TGR retinas at 1- and 3 months.A. Translational analysis of CD74 by Western blot. B. quantitative analysis of translational CD74 expression. The values at 1-month are standardized to 1. C: expression and localization of CD74 in the retina. Vessels are visualized by staining with Lectin (red), and CD74 positive cells are displayed in green. Few CD74 positive cells are seen in SD in the superficial and deep capillary layers. A slight increase in number of CD74 positive cells is found in TGR in the superficial capillary layer at 1 month. The numbers of CD74 positive cells are raised in the deep capillary layer in 3-month TGR compared with other three groups at all time points observed. C–F: the superficial capillary layer; G–J: the deep capillary layer. 1 m: 1-month retina. 3 m: 3-month retina. *p<0.05: 3-month TGR compared with other three groups.
Mentions: To investigate translational expression levels of CD74 obtained on the microarray, we performed western blot analysis with retinal lysates from TGR and SD rats at 1 and 3 months. Weak CD74 expression was found in the retinas of 1- and 3-month SD and 1-month TGR rats. A significant overexpression of CD74 protein was detected in 3-month TGR retinas compared with other three groups (Figure 5A and B).

Bottom Line: Of the 157 genes regulated more than twofold, the MHC class II invariant chain CD74 yielded the strongest upregulation, and was allocated to activated microglial cells close to the vessels undergoing vasoregression.Pathway clustering identified genes of the immune system including inflammatory signaling, and components of the complement cascade upregulated during vasoregression.Together, our data suggest that microglial cells involved in retinal immune response participate in the initiation of vasoregression in the retina.

View Article: PubMed Central - PubMed

Affiliation: 5 Medical Department, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany.

ABSTRACT
Vasoregression is a hallmark of vascular eye diseases but the mechanisms involved are still largely unknown. We have recently characterized a rat ciliopathy model which develops primary photoreceptor degeneration and secondary vasoregression. To improve the understanding of secondary vasoregression in retinal neurodegeneration, we used microarray techniques to compare gene expression profiles in this new model before and after retinal vasoregression. Differential gene expression was validated by quantitative RT-PCR, Western blot and immunofluorescence. Of the 157 genes regulated more than twofold, the MHC class II invariant chain CD74 yielded the strongest upregulation, and was allocated to activated microglial cells close to the vessels undergoing vasoregression. Pathway clustering identified genes of the immune system including inflammatory signaling, and components of the complement cascade upregulated during vasoregression. Together, our data suggest that microglial cells involved in retinal immune response participate in the initiation of vasoregression in the retina.

Show MeSH
Related in: MedlinePlus