Gene expression changes under cyclic mechanical stretching in rat retinal glial (Müller) cells
The retina is subjected to tractional forces in various conditions. As the predominant glial element in the retina, Müller cells are active players in all forms of retinal injury and disease. In this study, we aim to identify patterns of gene expression changes induced by cyclic mechanical stretchin...
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| description | The retina is subjected to tractional forces in various conditions. As the predominant glial element in the retina, Müller cells are active players in all forms of retinal injury and disease. In this study, we aim to identify patterns of gene expression changes induced by cyclic mechanical stretching in Müller cells.
Rat Müller cells were seeded onto flexible bottom culture plates and subjected to a cyclic stretching regimen of 15% equibiaxial stretching for 1 and 24 h. RNA was extracted and amplified, labeled, and hybridized to rat genome microarrays. The expression profiles were analyzed using GeneSpring software, and gene ontology analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to select, annotate, and visualize genes by function and pathway. The selected genes of interest were further validated by Quantitative Real-time PCR (qPCR).
Microarray data analysis showed that at 1 and 24 h, the expression of 532 and 991 genes in the Müller cells significantly (t-test, p |
| doi_str_mv | 10.1371/journal.pone.0063467 |
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Rat Müller cells were seeded onto flexible bottom culture plates and subjected to a cyclic stretching regimen of 15% equibiaxial stretching for 1 and 24 h. RNA was extracted and amplified, labeled, and hybridized to rat genome microarrays. The expression profiles were analyzed using GeneSpring software, and gene ontology analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to select, annotate, and visualize genes by function and pathway. The selected genes of interest were further validated by Quantitative Real-time PCR (qPCR).
Microarray data analysis showed that at 1 and 24 h, the expression of 532 and 991 genes in the Müller cells significantly (t-test, p<0.05) differed between the mechanically stretched and unstretched groups. Of these genes, 56 genes at 1 h and 62 genes at 24 h showed more than a twofold change in expression. Several genes related to response to stimulus (e.g., Egr2, IL6), cell proliferation (e.g., Areg, Atf3), tissue remodeling (e.g., PVR, Loxl2), and vasculogenesis (e.g., Epha2, Nrn1) were selected and validated by qPCR. KEGG pathway analysis showed significant changes in MAPK signaling at both time points.
Cyclic mechanical strain induces extensive changes in the gene expression in Müller cells through multiple molecular pathways. These results indicate the complex mechanoresponsive nature of Müller cells, and they provide novel insights into possible molecular mechanisms that would account for many retinal diseases in which the retina is often subjected to mechanical forces, such as pathological myopia and proliferative vitreoretinopathy.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0063467</identifier><identifier>PMID: 23723984</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Activating transcription factor 3 ; Animals ; Biology ; Biomechanical Phenomena ; Cell culture ; Cell growth ; Cell proliferation ; Cells, Cultured ; Cluster Analysis ; Data analysis ; Data processing ; DNA microarrays ; Egr-2 protein ; Encyclopedias ; Ependymoglial Cells - physiology ; EphA2 protein ; Gene expression ; Gene Expression Regulation ; Genes ; Genomes ; Hospitals ; Hybridization ; Interleukin 6 ; Laboratories ; MAP kinase ; Mechanical stimuli ; Medicine ; Metabolism ; Molecular modelling ; Myopia ; Oligonucleotide Array Sequence Analysis ; Protein Interaction Maps ; Rats ; Rats, Sprague-Dawley ; Retina ; Retina - cytology ; Ribonucleic acid ; RNA ; Signaling ; Smooth muscle ; Strain ; Stress, Physiological ; Stretching ; Transcriptome ; Visual impairment ; West Nile virus</subject><ispartof>PloS one, 2013-05, Vol.8 (5), p.e63467-e63467</ispartof><rights>2013 Wang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2013 Wang et al 2013 Wang et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c526t-37dcf098c46d4da2845f5c605eee63b8d75c0fb33073493e62dfbb9e833a29b53</citedby><cites>FETCH-LOGICAL-c526t-37dcf098c46d4da2845f5c605eee63b8d75c0fb33073493e62dfbb9e833a29b53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3664568/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3664568/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,724,777,781,861,882,2096,2915,23847,27905,27906,53772,53774,79349,79350</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23723984$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Rendon, Alvaro</contributor><creatorcontrib>Wang, Xin</creatorcontrib><creatorcontrib>Fan, Jiawen</creatorcontrib><creatorcontrib>Zhang, Meng</creatorcontrib><creatorcontrib>Sun, Zhongcui</creatorcontrib><creatorcontrib>Xu, Gezhi</creatorcontrib><title>Gene expression changes under cyclic mechanical stretching in rat retinal glial (Müller) cells</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>The retina is subjected to tractional forces in various conditions. As the predominant glial element in the retina, Müller cells are active players in all forms of retinal injury and disease. In this study, we aim to identify patterns of gene expression changes induced by cyclic mechanical stretching in Müller cells.
Rat Müller cells were seeded onto flexible bottom culture plates and subjected to a cyclic stretching regimen of 15% equibiaxial stretching for 1 and 24 h. RNA was extracted and amplified, labeled, and hybridized to rat genome microarrays. The expression profiles were analyzed using GeneSpring software, and gene ontology analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to select, annotate, and visualize genes by function and pathway. The selected genes of interest were further validated by Quantitative Real-time PCR (qPCR).
Microarray data analysis showed that at 1 and 24 h, the expression of 532 and 991 genes in the Müller cells significantly (t-test, p<0.05) differed between the mechanically stretched and unstretched groups. Of these genes, 56 genes at 1 h and 62 genes at 24 h showed more than a twofold change in expression. Several genes related to response to stimulus (e.g., Egr2, IL6), cell proliferation (e.g., Areg, Atf3), tissue remodeling (e.g., PVR, Loxl2), and vasculogenesis (e.g., Epha2, Nrn1) were selected and validated by qPCR. KEGG pathway analysis showed significant changes in MAPK signaling at both time points.
Cyclic mechanical strain induces extensive changes in the gene expression in Müller cells through multiple molecular pathways. These results indicate the complex mechanoresponsive nature of Müller cells, and they provide novel insights into possible molecular mechanisms that would account for many retinal diseases in which the retina is often subjected to mechanical forces, such as pathological myopia and proliferative vitreoretinopathy.</description><subject>Activating transcription factor 3</subject><subject>Animals</subject><subject>Biology</subject><subject>Biomechanical Phenomena</subject><subject>Cell culture</subject><subject>Cell growth</subject><subject>Cell proliferation</subject><subject>Cells, Cultured</subject><subject>Cluster Analysis</subject><subject>Data analysis</subject><subject>Data processing</subject><subject>DNA microarrays</subject><subject>Egr-2 protein</subject><subject>Encyclopedias</subject><subject>Ependymoglial Cells - physiology</subject><subject>EphA2 protein</subject><subject>Gene expression</subject><subject>Gene Expression Regulation</subject><subject>Genes</subject><subject>Genomes</subject><subject>Hospitals</subject><subject>Hybridization</subject><subject>Interleukin 6</subject><subject>Laboratories</subject><subject>MAP kinase</subject><subject>Mechanical stimuli</subject><subject>Medicine</subject><subject>Metabolism</subject><subject>Molecular modelling</subject><subject>Myopia</subject><subject>Oligonucleotide Array Sequence Analysis</subject><subject>Protein Interaction Maps</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>Retina</subject><subject>Retina - cytology</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>Signaling</subject><subject>Smooth muscle</subject><subject>Strain</subject><subject>Stress, Physiological</subject><subject>Stretching</subject><subject>Transcriptome</subject><subject>Visual impairment</subject><subject>West Nile virus</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNptUsFu1DAQjRCIlsIfILDEpRx2cTK241yQUAWlUhEXOFuOPcl65bUXO0H037j1x5p006pFXGzP-M2bN6NXFK9Lui6hLj9s45iC9ut9DLimVAAT9ZPiuGygWomKwtMH76PiRc5bSjlIIZ4XRxXUFTSSHRfqHAMS_LNPmLOLgZiNDj1mMgaLiZgr450hO5zTzmhP8pBwMBsXeuICSXogU-wmIaT3bjpPv13_9R7Te2LQ-_yyeNZpn_HVcp8UP798_nH2dXX5_fzi7NPlyvBKDCuoreloIw0TllldScY7bgTliCiglbbmhnYtAK2BNYCisl3bNigBdNW0HE6KtwfevY9ZLbvJqgTORSNELSfExQFho96qfXI7na5U1E7dJmLqlU6DMx5VIyuNVvCpJ2cNp5oZYYwseWfLKYcT18el29ju0BoMQ9L-Eenjn-A2qo-_FQjBuJjFnC4EKf4aMQ9q5_K8MB0wjre6a9ZIAWKCvvsH-v_p2AFlUsw5YXcvpqRq9stdlZr9oha_TGVvHg5yX3RnELgBj4q_6A</recordid><startdate>20130527</startdate><enddate>20130527</enddate><creator>Wang, Xin</creator><creator>Fan, Jiawen</creator><creator>Zhang, Meng</creator><creator>Sun, Zhongcui</creator><creator>Xu, Gezhi</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20130527</creationdate><title>Gene expression changes under cyclic mechanical stretching in rat retinal glial (Müller) cells</title><author>Wang, Xin ; Fan, Jiawen ; Zhang, Meng ; Sun, Zhongcui ; Xu, Gezhi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c526t-37dcf098c46d4da2845f5c605eee63b8d75c0fb33073493e62dfbb9e833a29b53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Activating transcription factor 3</topic><topic>Animals</topic><topic>Biology</topic><topic>Biomechanical Phenomena</topic><topic>Cell culture</topic><topic>Cell growth</topic><topic>Cell proliferation</topic><topic>Cells, Cultured</topic><topic>Cluster Analysis</topic><topic>Data analysis</topic><topic>Data processing</topic><topic>DNA microarrays</topic><topic>Egr-2 protein</topic><topic>Encyclopedias</topic><topic>Ependymoglial Cells - physiology</topic><topic>EphA2 protein</topic><topic>Gene expression</topic><topic>Gene Expression Regulation</topic><topic>Genes</topic><topic>Genomes</topic><topic>Hospitals</topic><topic>Hybridization</topic><topic>Interleukin 6</topic><topic>Laboratories</topic><topic>MAP kinase</topic><topic>Mechanical stimuli</topic><topic>Medicine</topic><topic>Metabolism</topic><topic>Molecular modelling</topic><topic>Myopia</topic><topic>Oligonucleotide Array Sequence Analysis</topic><topic>Protein Interaction Maps</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>Retina</topic><topic>Retina - cytology</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>Signaling</topic><topic>Smooth muscle</topic><topic>Strain</topic><topic>Stress, Physiological</topic><topic>Stretching</topic><topic>Transcriptome</topic><topic>Visual impairment</topic><topic>West Nile virus</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Xin</creatorcontrib><creatorcontrib>Fan, Jiawen</creatorcontrib><creatorcontrib>Zhang, Meng</creatorcontrib><creatorcontrib>Sun, Zhongcui</creatorcontrib><creatorcontrib>Xu, Gezhi</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Xin</au><au>Fan, Jiawen</au><au>Zhang, Meng</au><au>Sun, Zhongcui</au><au>Xu, Gezhi</au><au>Rendon, Alvaro</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Gene expression changes under cyclic mechanical stretching in rat retinal glial (Müller) cells</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2013-05-27</date><risdate>2013</risdate><volume>8</volume><issue>5</issue><spage>e63467</spage><epage>e63467</epage><pages>e63467-e63467</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>The retina is subjected to tractional forces in various conditions. As the predominant glial element in the retina, Müller cells are active players in all forms of retinal injury and disease. In this study, we aim to identify patterns of gene expression changes induced by cyclic mechanical stretching in Müller cells.
Rat Müller cells were seeded onto flexible bottom culture plates and subjected to a cyclic stretching regimen of 15% equibiaxial stretching for 1 and 24 h. RNA was extracted and amplified, labeled, and hybridized to rat genome microarrays. The expression profiles were analyzed using GeneSpring software, and gene ontology analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to select, annotate, and visualize genes by function and pathway. The selected genes of interest were further validated by Quantitative Real-time PCR (qPCR).
Microarray data analysis showed that at 1 and 24 h, the expression of 532 and 991 genes in the Müller cells significantly (t-test, p<0.05) differed between the mechanically stretched and unstretched groups. Of these genes, 56 genes at 1 h and 62 genes at 24 h showed more than a twofold change in expression. Several genes related to response to stimulus (e.g., Egr2, IL6), cell proliferation (e.g., Areg, Atf3), tissue remodeling (e.g., PVR, Loxl2), and vasculogenesis (e.g., Epha2, Nrn1) were selected and validated by qPCR. KEGG pathway analysis showed significant changes in MAPK signaling at both time points.
Cyclic mechanical strain induces extensive changes in the gene expression in Müller cells through multiple molecular pathways. These results indicate the complex mechanoresponsive nature of Müller cells, and they provide novel insights into possible molecular mechanisms that would account for many retinal diseases in which the retina is often subjected to mechanical forces, such as pathological myopia and proliferative vitreoretinopathy.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23723984</pmid><doi>10.1371/journal.pone.0063467</doi><oa>free_for_read</oa></addata></record> |
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| subjects | Activating transcription factor 3 Animals Biology Biomechanical Phenomena Cell culture Cell growth Cell proliferation Cells, Cultured Cluster Analysis Data analysis Data processing DNA microarrays Egr-2 protein Encyclopedias Ependymoglial Cells - physiology EphA2 protein Gene expression Gene Expression Regulation Genes Genomes Hospitals Hybridization Interleukin 6 Laboratories MAP kinase Mechanical stimuli Medicine Metabolism Molecular modelling Myopia Oligonucleotide Array Sequence Analysis Protein Interaction Maps Rats Rats, Sprague-Dawley Retina Retina - cytology Ribonucleic acid RNA Signaling Smooth muscle Strain Stress, Physiological Stretching Transcriptome Visual impairment West Nile virus |
| title | Gene expression changes under cyclic mechanical stretching in rat retinal glial (Müller) cells |
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