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Ciliary neurotrophic factor induces genes associated with inflammation and gliosis in the retina: a gene profiling study of flow-sorted, Müller cells.

Xue W, Cojocaru RI, Dudley VJ, Brooks M, Swaroop A, Sarthy VP - PLoS ONE (2011)

Bottom Line: A comparison of transcriptional profiles from Müller cells at one or three days after CNTF treatment showed an increase in the number of transcribed genes as well as a change in the expression pattern.Further analysis of gene profiles revealed 20-30% overlap in the transcription pattern among Müller cells, astrocytes and the RPE.Our studies provide novel molecular insights into biological functions of Müller glial cells in mediating cytokine response.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Northwestern University Feinberg Medical School, Chicago, Illinois, United States of America.

ABSTRACT

Background: Ciliary neurotrophic factor (CNTF), a member of the interleukin-6 cytokine family, has been implicated in the development, differentiation and survival of retinal neurons. The mechanisms of CNTF action as well as its cellular targets in the retina are poorly understood. It has been postulated that some of the biological effects of CNTF are mediated through its action via retinal glial cells; however, molecular changes in retinal glia induced by CNTF have not been elucidated. We have, therefore, examined gene expression dynamics of purified Müller (glial) cells exposed to CNTF in vivo.

Methodology/principal findings: Müller cells were flow-sorted from mgfap-egfp transgenic mice one or three days after intravitreal injection of CNTF. Microarray analysis using RNA from purified Müller cells showed differential expression of almost 1,000 transcripts with two- to seventeen-fold change in response to CNTF. A comparison of transcriptional profiles from Müller cells at one or three days after CNTF treatment showed an increase in the number of transcribed genes as well as a change in the expression pattern. Ingenuity Pathway Analysis showed that the differentially regulated genes belong to distinct functional types such as cytokines, growth factors, G-protein coupled receptors, transporters and ion channels. Interestingly, many genes induced by CNTF were also highly expressed in reactive Müller cells from mice with inherited or experimentally induced retinal degeneration. Further analysis of gene profiles revealed 20-30% overlap in the transcription pattern among Müller cells, astrocytes and the RPE.

Conclusions/significance: Our studies provide novel molecular insights into biological functions of Müller glial cells in mediating cytokine response. We suggest that CNTF remodels the gene expression profile of Müller cells leading to induction of networks associated with transcription, cell cycle regulation and inflammatory response. CNTF also appears to function as an inducer of gliosis in the retina.

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Microarray data analysis.(a) Hierarchical clustering showing 1261 probes that have a P-val ≤0.1 and a minimum 2-fold change between the CNTF and PBS samples at Day 1. Bright blue indicates lowest signal with increasing values indicated by yellow shading to bright red, representing peak signal. (b) Most significant 10 canonical pathways corresponding to Day 1. (c) Most significant biological functions for the same list of genes. (d) Hierarchical clustering showing 1541 probes that have a P-val ≤0.1 and a minimum 2-fold change between the CNTF and PBS samples at Day 3. (e) Most significant 10 canonical pathways corresponding to Day 3. (f) Most significant biological functions for the same list of genes.
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pone-0020326-g002: Microarray data analysis.(a) Hierarchical clustering showing 1261 probes that have a P-val ≤0.1 and a minimum 2-fold change between the CNTF and PBS samples at Day 1. Bright blue indicates lowest signal with increasing values indicated by yellow shading to bright red, representing peak signal. (b) Most significant 10 canonical pathways corresponding to Day 1. (c) Most significant biological functions for the same list of genes. (d) Hierarchical clustering showing 1541 probes that have a P-val ≤0.1 and a minimum 2-fold change between the CNTF and PBS samples at Day 3. (e) Most significant 10 canonical pathways corresponding to Day 3. (f) Most significant biological functions for the same list of genes.

Mentions: To study the transcriptional response of Müller cells to CNTF, mgfap-egfp transgenic mice were intravitreally injected with CNTF or PBS (control). One day later, GFP+-Müller cells were flow-sorted, and total RNA was prepared for microarray analysis [49]. CNTF treatment resulted in differential expression of 923 transcripts that showed at least two-fold change (P-value <0.05). Of these, 691 transcripts exhibited 2-to 17-fold higher expression, whereas 232 transcripts revealed 2-to 5-fold reduction. We noticed, however, that genes such as gfap that are known to be induced by CNTF, were not detected. When the microarray data were analyzed with P-value<0.1, however, we found several genes of biological interest among 1261 differentially expressed transcripts. Of these, 939 transcripts were elevated 2- to 17-fold, and 322 transcripts were depressed 2- to 5-fold. The differentially expressed genes fell into several functional types (Table 1). A complete list of differentially regulated genes is presented in Table S1. A hierarchical cluster analysis also showed that genes with increased expression level outnumbered genes with decreased level, and that CNTF-induced genes fell into several clusters of co-expressed genes (Figure 2).


Ciliary neurotrophic factor induces genes associated with inflammation and gliosis in the retina: a gene profiling study of flow-sorted, Müller cells.

Xue W, Cojocaru RI, Dudley VJ, Brooks M, Swaroop A, Sarthy VP - PLoS ONE (2011)

Microarray data analysis.(a) Hierarchical clustering showing 1261 probes that have a P-val ≤0.1 and a minimum 2-fold change between the CNTF and PBS samples at Day 1. Bright blue indicates lowest signal with increasing values indicated by yellow shading to bright red, representing peak signal. (b) Most significant 10 canonical pathways corresponding to Day 1. (c) Most significant biological functions for the same list of genes. (d) Hierarchical clustering showing 1541 probes that have a P-val ≤0.1 and a minimum 2-fold change between the CNTF and PBS samples at Day 3. (e) Most significant 10 canonical pathways corresponding to Day 3. (f) Most significant biological functions for the same list of genes.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3102695&req=5

pone-0020326-g002: Microarray data analysis.(a) Hierarchical clustering showing 1261 probes that have a P-val ≤0.1 and a minimum 2-fold change between the CNTF and PBS samples at Day 1. Bright blue indicates lowest signal with increasing values indicated by yellow shading to bright red, representing peak signal. (b) Most significant 10 canonical pathways corresponding to Day 1. (c) Most significant biological functions for the same list of genes. (d) Hierarchical clustering showing 1541 probes that have a P-val ≤0.1 and a minimum 2-fold change between the CNTF and PBS samples at Day 3. (e) Most significant 10 canonical pathways corresponding to Day 3. (f) Most significant biological functions for the same list of genes.
Mentions: To study the transcriptional response of Müller cells to CNTF, mgfap-egfp transgenic mice were intravitreally injected with CNTF or PBS (control). One day later, GFP+-Müller cells were flow-sorted, and total RNA was prepared for microarray analysis [49]. CNTF treatment resulted in differential expression of 923 transcripts that showed at least two-fold change (P-value <0.05). Of these, 691 transcripts exhibited 2-to 17-fold higher expression, whereas 232 transcripts revealed 2-to 5-fold reduction. We noticed, however, that genes such as gfap that are known to be induced by CNTF, were not detected. When the microarray data were analyzed with P-value<0.1, however, we found several genes of biological interest among 1261 differentially expressed transcripts. Of these, 939 transcripts were elevated 2- to 17-fold, and 322 transcripts were depressed 2- to 5-fold. The differentially expressed genes fell into several functional types (Table 1). A complete list of differentially regulated genes is presented in Table S1. A hierarchical cluster analysis also showed that genes with increased expression level outnumbered genes with decreased level, and that CNTF-induced genes fell into several clusters of co-expressed genes (Figure 2).

Bottom Line: A comparison of transcriptional profiles from Müller cells at one or three days after CNTF treatment showed an increase in the number of transcribed genes as well as a change in the expression pattern.Further analysis of gene profiles revealed 20-30% overlap in the transcription pattern among Müller cells, astrocytes and the RPE.Our studies provide novel molecular insights into biological functions of Müller glial cells in mediating cytokine response.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Northwestern University Feinberg Medical School, Chicago, Illinois, United States of America.

ABSTRACT

Background: Ciliary neurotrophic factor (CNTF), a member of the interleukin-6 cytokine family, has been implicated in the development, differentiation and survival of retinal neurons. The mechanisms of CNTF action as well as its cellular targets in the retina are poorly understood. It has been postulated that some of the biological effects of CNTF are mediated through its action via retinal glial cells; however, molecular changes in retinal glia induced by CNTF have not been elucidated. We have, therefore, examined gene expression dynamics of purified Müller (glial) cells exposed to CNTF in vivo.

Methodology/principal findings: Müller cells were flow-sorted from mgfap-egfp transgenic mice one or three days after intravitreal injection of CNTF. Microarray analysis using RNA from purified Müller cells showed differential expression of almost 1,000 transcripts with two- to seventeen-fold change in response to CNTF. A comparison of transcriptional profiles from Müller cells at one or three days after CNTF treatment showed an increase in the number of transcribed genes as well as a change in the expression pattern. Ingenuity Pathway Analysis showed that the differentially regulated genes belong to distinct functional types such as cytokines, growth factors, G-protein coupled receptors, transporters and ion channels. Interestingly, many genes induced by CNTF were also highly expressed in reactive Müller cells from mice with inherited or experimentally induced retinal degeneration. Further analysis of gene profiles revealed 20-30% overlap in the transcription pattern among Müller cells, astrocytes and the RPE.

Conclusions/significance: Our studies provide novel molecular insights into biological functions of Müller glial cells in mediating cytokine response. We suggest that CNTF remodels the gene expression profile of Müller cells leading to induction of networks associated with transcription, cell cycle regulation and inflammatory response. CNTF also appears to function as an inducer of gliosis in the retina.

Show MeSH
Related in: MedlinePlus