Silencing of GPNMB by siRNA inhibits the formation of melanosomes in melanocytes in a MITF-independent fashion
Melanosomes are specialized membrane-surrounded organelles, which are involved in the synthesis, storage and transport of melanin. Glycoprotein (transmembrane) non-metastatic melanoma protein b (GPNMB), a melanosome-specific structural protein, shares significant amino acid sequence homology with Pm...
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| description | Melanosomes are specialized membrane-surrounded organelles, which are involved in the synthesis, storage and transport of melanin. Glycoprotein (transmembrane) non-metastatic melanoma protein b (GPNMB), a melanosome-specific structural protein, shares significant amino acid sequence homology with Pmel-17. Proteomic analysis demonstrated that GPNMB is present in all stages (I-IV) of melanosomes. However, little is known about the role of GPNMB in melanosomes.
Using real-time quantitative PCR, Western blotting and immunofluorescence analysis, we demonstrated that the expression of GPNMB in PIG1 melanocytes was up-regulated by ultraviolet B (UVB) radiation. Transmission electron microscopy analysis showed that the total number of melanosomes in PIG1 melanocytes was sharply reduced by GPNMB-siRNA transfection. Simultaneously, the expression levels of tyrosinase (Tyr), tyrosinase related protein 1 (Trp1), Pmel17/gp100 and ocular albinism type 1 protein (OA1) were all significantly attenuated. But the expression of microphthalmia-associated transcription factor (MITF) was up-regulated. Intriguingly, in GPNMB silenced PIG1 melanocytes, UVB radiation sharply reduced MITF expression.
Our present work revealed that the GPNMB was critical for the formation of melanosomes. And GPNMB expression down-regulation attenuated melanosome formation in a MITF-independent fashion. |
| doi_str_mv | 10.1371/journal.pone.0042955 |
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Using real-time quantitative PCR, Western blotting and immunofluorescence analysis, we demonstrated that the expression of GPNMB in PIG1 melanocytes was up-regulated by ultraviolet B (UVB) radiation. Transmission electron microscopy analysis showed that the total number of melanosomes in PIG1 melanocytes was sharply reduced by GPNMB-siRNA transfection. Simultaneously, the expression levels of tyrosinase (Tyr), tyrosinase related protein 1 (Trp1), Pmel17/gp100 and ocular albinism type 1 protein (OA1) were all significantly attenuated. But the expression of microphthalmia-associated transcription factor (MITF) was up-regulated. Intriguingly, in GPNMB silenced PIG1 melanocytes, UVB radiation sharply reduced MITF expression.
Our present work revealed that the GPNMB was critical for the formation of melanosomes. And GPNMB expression down-regulation attenuated melanosome formation in a MITF-independent fashion.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0042955</identifier><identifier>PMID: 22912767</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Albinism ; Amino acid sequence ; Amino acids ; Analysis ; Biology ; Blotting, Western ; Cancer metastasis ; Cell Line ; Dermatology ; DNA Primers - genetics ; Electron microscopy ; Eye Proteins - metabolism ; Flow Cytometry ; Gene expression ; Gene Expression Regulation - radiation effects ; Gene Silencing ; Glycoprotein gp100 ; Glycoproteins ; Homology ; Humans ; Immunofluorescence ; Medicine ; Melanin ; Melanocytes ; Melanocytes - metabolism ; Melanoma ; Melanosomes ; Melanosomes - metabolism ; Membrane Glycoproteins - genetics ; Membrane Glycoproteins - metabolism ; Metastases ; Metastasis ; Microphthalmia-associated transcription factor ; Microphthalmia-Associated Transcription Factor - metabolism ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; Monophenol Monooxygenase - metabolism ; Ocular albinism ; Organelles ; Protein B ; Protein expression ; Proteins ; Proteomics ; Radiation ; Radiation (Physics) ; Real-Time Polymerase Chain Reaction ; RNA, Small Interfering - genetics ; siRNA ; Transcription factors ; Transfection ; Transmission electron microscopy ; Trypsin - metabolism ; Tyrosinase ; U.V. radiation ; Ultraviolet radiation ; Ultraviolet Rays ; Western blotting</subject><ispartof>PloS one, 2012-08, Vol.7 (8), p.e42955</ispartof><rights>COPYRIGHT 2012 Public Library of Science</rights><rights>Zhang 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>2012 Zhang et al 2012 Zhang et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-5f080571f27a48c72983833106e3eb19bbf515e03d219210a2164be9f8ae78123</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3418242/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3418242/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,315,728,781,785,865,886,2103,2929,23871,27929,27930,53796,53798</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22912767$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Veitia, Reiner Albert</contributor><creatorcontrib>Zhang, Ping</creatorcontrib><creatorcontrib>Liu, Wei</creatorcontrib><creatorcontrib>Zhu, Cansheng</creatorcontrib><creatorcontrib>Yuan, Xiaoying</creatorcontrib><creatorcontrib>Li, Dongguang</creatorcontrib><creatorcontrib>Gu, Weijie</creatorcontrib><creatorcontrib>Ma, Huimin</creatorcontrib><creatorcontrib>Xie, Xin</creatorcontrib><creatorcontrib>Gao, Tianwen</creatorcontrib><title>Silencing of GPNMB by siRNA inhibits the formation of melanosomes in melanocytes in a MITF-independent fashion</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Melanosomes are specialized membrane-surrounded organelles, which are involved in the synthesis, storage and transport of melanin. Glycoprotein (transmembrane) non-metastatic melanoma protein b (GPNMB), a melanosome-specific structural protein, shares significant amino acid sequence homology with Pmel-17. Proteomic analysis demonstrated that GPNMB is present in all stages (I-IV) of melanosomes. However, little is known about the role of GPNMB in melanosomes.
Using real-time quantitative PCR, Western blotting and immunofluorescence analysis, we demonstrated that the expression of GPNMB in PIG1 melanocytes was up-regulated by ultraviolet B (UVB) radiation. Transmission electron microscopy analysis showed that the total number of melanosomes in PIG1 melanocytes was sharply reduced by GPNMB-siRNA transfection. Simultaneously, the expression levels of tyrosinase (Tyr), tyrosinase related protein 1 (Trp1), Pmel17/gp100 and ocular albinism type 1 protein (OA1) were all significantly attenuated. But the expression of microphthalmia-associated transcription factor (MITF) was up-regulated. Intriguingly, in GPNMB silenced PIG1 melanocytes, UVB radiation sharply reduced MITF expression.
Our present work revealed that the GPNMB was critical for the formation of melanosomes. And GPNMB expression down-regulation attenuated melanosome formation in a MITF-independent fashion.</description><subject>Albinism</subject><subject>Amino acid sequence</subject><subject>Amino acids</subject><subject>Analysis</subject><subject>Biology</subject><subject>Blotting, Western</subject><subject>Cancer metastasis</subject><subject>Cell Line</subject><subject>Dermatology</subject><subject>DNA Primers - genetics</subject><subject>Electron microscopy</subject><subject>Eye Proteins - metabolism</subject><subject>Flow Cytometry</subject><subject>Gene expression</subject><subject>Gene Expression Regulation - radiation effects</subject><subject>Gene Silencing</subject><subject>Glycoprotein gp100</subject><subject>Glycoproteins</subject><subject>Homology</subject><subject>Humans</subject><subject>Immunofluorescence</subject><subject>Medicine</subject><subject>Melanin</subject><subject>Melanocytes</subject><subject>Melanocytes - metabolism</subject><subject>Melanoma</subject><subject>Melanosomes</subject><subject>Melanosomes - metabolism</subject><subject>Membrane Glycoproteins - genetics</subject><subject>Membrane Glycoproteins - metabolism</subject><subject>Metastases</subject><subject>Metastasis</subject><subject>Microphthalmia-associated transcription factor</subject><subject>Microphthalmia-Associated Transcription Factor - metabolism</subject><subject>Microscopy, Electron, Transmission</subject><subject>Microscopy, Fluorescence</subject><subject>Monophenol Monooxygenase - metabolism</subject><subject>Ocular albinism</subject><subject>Organelles</subject><subject>Protein B</subject><subject>Protein expression</subject><subject>Proteins</subject><subject>Proteomics</subject><subject>Radiation</subject><subject>Radiation (Physics)</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>RNA, Small Interfering - genetics</subject><subject>siRNA</subject><subject>Transcription factors</subject><subject>Transfection</subject><subject>Transmission electron microscopy</subject><subject>Trypsin - metabolism</subject><subject>Tyrosinase</subject><subject>U.V. radiation</subject><subject>Ultraviolet radiation</subject><subject>Ultraviolet Rays</subject><subject>Western blotting</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</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>eNqNkl2L1DAYhYso7rr6D0QLguDFjPlqk94sjIu7DuyH7K7ehiRNphnaZGwy4vx7U6e7TEFBAk3TPOfk7ZuTZa8hmENM4ce13_ZOtPONd3oOAEFVUTzJjmGF0axEAD89eD_KXoSwBqDArCyfZ0cIVRDRkh5n7s622inrVrk3-cXX66tPudzlwd5eL3LrGittDHlsdG5834lovRvATrfC-eA7HRI1LtUu7pciv1ren8-sq_VGp4eLuRGhSdqX2TMj2qBfjfNJ9u388_3Zl9nlzcXybHE5U7RgcVYYwEBBoUFUEKYoqhhmGENQaqwlrKQ0BSw0wDWCFYJAIFgSqSvDhKYMInySvd37blof-NiqwCFGJSKQgDIRyz1Re7Hmm952ot9xLyz_88H3Ky76aFWruRIMFzUkqpCEoBJLpTSSRjJkhqNp8jodT9vKTtcq_XAv2onpdMfZhq_8T44JZIgM5b4bDXr_Y6tD_EfJI7USqSrrjE9mqrNB8UVBMAYU0qGY-V-oNGrdWZXCYtKNTwUfJoLERP0rrsQ2BL68u_1_9ub7lH1_wDZatLEJvt0OGQpTkOxB1fsQem0eOwcBH7L-0A0-ZJ2PWU-yN4ddfxQ9hBv_Bi7--Bk</recordid><startdate>20120813</startdate><enddate>20120813</enddate><creator>Zhang, Ping</creator><creator>Liu, Wei</creator><creator>Zhu, Cansheng</creator><creator>Yuan, Xiaoying</creator><creator>Li, Dongguang</creator><creator>Gu, Weijie</creator><creator>Ma, Huimin</creator><creator>Xie, Xin</creator><creator>Gao, Tianwen</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>IOV</scope><scope>ISR</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>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>5PM</scope><scope>DOA</scope></search><sort><creationdate>20120813</creationdate><title>Silencing of GPNMB by siRNA inhibits the formation of melanosomes in melanocytes in a MITF-independent fashion</title><author>Zhang, Ping ; Liu, Wei ; Zhu, Cansheng ; Yuan, Xiaoying ; Li, Dongguang ; Gu, Weijie ; Ma, Huimin ; Xie, Xin ; Gao, Tianwen</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c758t-5f080571f27a48c72983833106e3eb19bbf515e03d219210a2164be9f8ae78123</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Albinism</topic><topic>Amino acid sequence</topic><topic>Amino acids</topic><topic>Analysis</topic><topic>Biology</topic><topic>Blotting, Western</topic><topic>Cancer metastasis</topic><topic>Cell Line</topic><topic>Dermatology</topic><topic>DNA Primers - genetics</topic><topic>Electron microscopy</topic><topic>Eye Proteins - metabolism</topic><topic>Flow Cytometry</topic><topic>Gene expression</topic><topic>Gene Expression Regulation - radiation effects</topic><topic>Gene Silencing</topic><topic>Glycoprotein gp100</topic><topic>Glycoproteins</topic><topic>Homology</topic><topic>Humans</topic><topic>Immunofluorescence</topic><topic>Medicine</topic><topic>Melanin</topic><topic>Melanocytes</topic><topic>Melanocytes - metabolism</topic><topic>Melanoma</topic><topic>Melanosomes</topic><topic>Melanosomes - metabolism</topic><topic>Membrane Glycoproteins - genetics</topic><topic>Membrane Glycoproteins - metabolism</topic><topic>Metastases</topic><topic>Metastasis</topic><topic>Microphthalmia-associated transcription factor</topic><topic>Microphthalmia-Associated Transcription Factor - metabolism</topic><topic>Microscopy, Electron, Transmission</topic><topic>Microscopy, Fluorescence</topic><topic>Monophenol Monooxygenase - metabolism</topic><topic>Ocular albinism</topic><topic>Organelles</topic><topic>Protein B</topic><topic>Protein expression</topic><topic>Proteins</topic><topic>Proteomics</topic><topic>Radiation</topic><topic>Radiation (Physics)</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>RNA, Small Interfering - genetics</topic><topic>siRNA</topic><topic>Transcription factors</topic><topic>Transfection</topic><topic>Transmission electron microscopy</topic><topic>Trypsin - metabolism</topic><topic>Tyrosinase</topic><topic>U.V. radiation</topic><topic>Ultraviolet radiation</topic><topic>Ultraviolet Rays</topic><topic>Western blotting</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhang, Ping</creatorcontrib><creatorcontrib>Liu, Wei</creatorcontrib><creatorcontrib>Zhu, Cansheng</creatorcontrib><creatorcontrib>Yuan, Xiaoying</creatorcontrib><creatorcontrib>Li, Dongguang</creatorcontrib><creatorcontrib>Gu, Weijie</creatorcontrib><creatorcontrib>Ma, Huimin</creatorcontrib><creatorcontrib>Xie, Xin</creatorcontrib><creatorcontrib>Gao, Tianwen</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</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 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 (ProQuest)</collection><collection>Natural Science Collection (ProQuest)</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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Glycoprotein (transmembrane) non-metastatic melanoma protein b (GPNMB), a melanosome-specific structural protein, shares significant amino acid sequence homology with Pmel-17. Proteomic analysis demonstrated that GPNMB is present in all stages (I-IV) of melanosomes. However, little is known about the role of GPNMB in melanosomes.
Using real-time quantitative PCR, Western blotting and immunofluorescence analysis, we demonstrated that the expression of GPNMB in PIG1 melanocytes was up-regulated by ultraviolet B (UVB) radiation. Transmission electron microscopy analysis showed that the total number of melanosomes in PIG1 melanocytes was sharply reduced by GPNMB-siRNA transfection. Simultaneously, the expression levels of tyrosinase (Tyr), tyrosinase related protein 1 (Trp1), Pmel17/gp100 and ocular albinism type 1 protein (OA1) were all significantly attenuated. But the expression of microphthalmia-associated transcription factor (MITF) was up-regulated. Intriguingly, in GPNMB silenced PIG1 melanocytes, UVB radiation sharply reduced MITF expression.
Our present work revealed that the GPNMB was critical for the formation of melanosomes. And GPNMB expression down-regulation attenuated melanosome formation in a MITF-independent fashion.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>22912767</pmid><doi>10.1371/journal.pone.0042955</doi><tpages>e42955</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | PloS one, 2012-08, Vol.7 (8), p.e42955 |
| issn | 1932-6203 1932-6203 |
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| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Albinism Amino acid sequence Amino acids Analysis Biology Blotting, Western Cancer metastasis Cell Line Dermatology DNA Primers - genetics Electron microscopy Eye Proteins - metabolism Flow Cytometry Gene expression Gene Expression Regulation - radiation effects Gene Silencing Glycoprotein gp100 Glycoproteins Homology Humans Immunofluorescence Medicine Melanin Melanocytes Melanocytes - metabolism Melanoma Melanosomes Melanosomes - metabolism Membrane Glycoproteins - genetics Membrane Glycoproteins - metabolism Metastases Metastasis Microphthalmia-associated transcription factor Microphthalmia-Associated Transcription Factor - metabolism Microscopy, Electron, Transmission Microscopy, Fluorescence Monophenol Monooxygenase - metabolism Ocular albinism Organelles Protein B Protein expression Proteins Proteomics Radiation Radiation (Physics) Real-Time Polymerase Chain Reaction RNA, Small Interfering - genetics siRNA Transcription factors Transfection Transmission electron microscopy Trypsin - metabolism Tyrosinase U.V. radiation Ultraviolet radiation Ultraviolet Rays Western blotting |
| title | Silencing of GPNMB by siRNA inhibits the formation of melanosomes in melanocytes in a MITF-independent fashion |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-13T04%3A07%3A05IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Silencing%20of%20GPNMB%20by%20siRNA%20inhibits%20the%20formation%20of%20melanosomes%20in%20melanocytes%20in%20a%20MITF-independent%20fashion&rft.jtitle=PloS%20one&rft.au=Zhang,%20Ping&rft.date=2012-08-13&rft.volume=7&rft.issue=8&rft.spage=e42955&rft.pages=e42955-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0042955&rft_dat=%3Cgale_plos_%3EA543307177%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1326241406&rft_id=info:pmid/22912767&rft_galeid=A543307177&rft_doaj_id=oai_doaj_org_article_ca835d14c5b44263bcce2bfb82f10a27&rfr_iscdi=true |