Human Pro370Leu Mutant Myocilin Induces the Phenotype of Open-Angle Glaucoma in Transgenic Mice
To investigate the characteristics of mutation myocilin proteins and glaucoma pathological phenotype in transgenic mice with full-length human Pro370Leu mutant myocilin gene (Tg- MYOC P370L ). Tg- MYOC P370L mice were established using the CRISPR/Cas9 system. Long-term intraocular pressure (IOP) was...
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| Veröffentlicht in: | Cellular and molecular neurobiology 2023-07, Vol.43 (5), p.2021-2033 |
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| creator | Cheng, Ying Wu, Shen Yan, Xuejing Liu, Qian Lin, Danting Zhang, Jingxue Wang, Ningli |
| description | To investigate the characteristics of mutation myocilin proteins and glaucoma pathological phenotype in transgenic mice with full-length human Pro370Leu mutant myocilin gene (Tg-
MYOC
P370L
). Tg-
MYOC
P370L
mice were established using the CRISPR/Cas9 system. Long-term intraocular pressure (IOP) was measured, myocilin protein expressions in anterior chamber angle, retina, optic nerve tissues and aqueous humor were detected by western blot. RBPMS, myocilin, Iba-1 and GFAP expression were visualized by immunofluorescence. H&E staining was applied to assess the ocular angle and retinal morphology. Aqueous humor dynamics were visualized by Gadolinium magnetic resonance imaging (Gd-MRI). TUNEL assay was used to evaluate the specific cell apoptosis in trabecular meshwork and retina. Optomotor and electroretinography tests were employed to evaluate the visual function in Tg-
MYOC
P370L
and wild-type (WT) mice. Homozygous myocilin mutation at position 503 (C > T) was identified by PCR and sequencing in Tg-
MYOC
P370L
mice. Myocilin protein expression was overexpressed in eye tissues of Tg-
MYOC
P370L
mice with reduced myocilin secretion in aqueous humor. H&E staining showed normal histological morphology of anterior chamber angle whereas decreased thickness and nuclei in ganglion cell layer were found (
P
|
| doi_str_mv | 10.1007/s10571-022-01280-x |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2711307515</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2828889645</sourcerecordid><originalsourceid>FETCH-LOGICAL-c375t-3afb21f4d9f2055914fbfa0cb4f4a7dd179aa2ac804f90e91aafad68bd5d95023</originalsourceid><addsrcrecordid>eNp9kLFuFDEQhi0EIpfAC1AgSzQ0hrG9XttlFIUk0p2SItTG67UvG-3ah72Wcm_Ds_Bk2XABJAqqKeb7_xl9CL2j8IkCyM-FgpCUAGMEKFNAHl6gFRWSk1ZxeIlWwCQjDW_gCB2Xcg8AGkC8Rke8hVZroVbo22WdbMQ3OXEJa1_xps42znizT24Yh4ivYl-dL3i-8_jmzsc073f-548U8PXOR3Iat6PHF6OtLk0WL4HbbGPZ-jg4vBmcf4NeBTsW__Z5nqCvX85vzy7J-vri6ux0TRyXYibcho7R0PQ6MBBC0yZ0wYLrmtBY2fdUamuZdQqaoMFram2wfau6XvRaAOMn6OOhd5fT9-rLbKahOD-ONvpUi2GSUg5SULGgH_5B71PNcfnOMMWUUrptnih2oFxOpWQfzC4Pk817Q8E8-TcH_2bxb375Nw9L6P1zde0m3_-J_Ba-APwAlGUVtz7_vf2f2kcNxpFF</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2828889645</pqid></control><display><type>article</type><title>Human Pro370Leu Mutant Myocilin Induces the Phenotype of Open-Angle Glaucoma in Transgenic Mice</title><source>SpringerLink Journals - AutoHoldings</source><creator>Cheng, Ying ; Wu, Shen ; Yan, Xuejing ; Liu, Qian ; Lin, Danting ; Zhang, Jingxue ; Wang, Ningli</creator><creatorcontrib>Cheng, Ying ; Wu, Shen ; Yan, Xuejing ; Liu, Qian ; Lin, Danting ; Zhang, Jingxue ; Wang, Ningli</creatorcontrib><description>To investigate the characteristics of mutation myocilin proteins and glaucoma pathological phenotype in transgenic mice with full-length human Pro370Leu mutant myocilin gene (Tg-
MYOC
P370L
). Tg-
MYOC
P370L
mice were established using the CRISPR/Cas9 system. Long-term intraocular pressure (IOP) was measured, myocilin protein expressions in anterior chamber angle, retina, optic nerve tissues and aqueous humor were detected by western blot. RBPMS, myocilin, Iba-1 and GFAP expression were visualized by immunofluorescence. H&E staining was applied to assess the ocular angle and retinal morphology. Aqueous humor dynamics were visualized by Gadolinium magnetic resonance imaging (Gd-MRI). TUNEL assay was used to evaluate the specific cell apoptosis in trabecular meshwork and retina. Optomotor and electroretinography tests were employed to evaluate the visual function in Tg-
MYOC
P370L
and wild-type (WT) mice. Homozygous myocilin mutation at position 503 (C > T) was identified by PCR and sequencing in Tg-
MYOC
P370L
mice. Myocilin protein expression was overexpressed in eye tissues of Tg-
MYOC
P370L
mice with reduced myocilin secretion in aqueous humor. H&E staining showed normal histological morphology of anterior chamber angle whereas decreased thickness and nuclei in ganglion cell layer were found (
P
< 0.05). Gd signals were significantly increased in the anterior chamber of Tg-
MYOC
P370L
compared with WT eyes (
P
< 0.05). IOP was elevated in Tg-
MYOC
P370L
mice starting at 5 months of age, with significant RGC loss (
P
< 0.05). Upregulation of caspase-3 and caspase-9 expressions and increased TUNEL-positive cells were found in eyes of Tg-
MYOC
P370L
mice. Excessive activation of retinal glial cells and impaired visual function were detected in Tg-
MYOC
P370L
mice. Tg-
MYOC
P370L
mice can induce the phenotype of open-angle glaucoma, featured as IOP elevation, activated retinal glial cells, loss of RGCs and impaired visual function. These pathologic changes may arise from the abnormal mutant myocilin protein accumulation in the trabecular meshwork and injured aqueous humor drainage. Therefore, Tg-
MYOC
P370L
mice model can serve as an effective animal model for glaucoma research, especially for glaucoma-associated myocilin mutation studies.</description><identifier>ISSN: 0272-4340</identifier><identifier>EISSN: 1573-6830</identifier><identifier>DOI: 10.1007/s10571-022-01280-x</identifier><identifier>PMID: 36069958</identifier><language>eng</language><publisher>New York: Springer US</publisher><subject>Animal models ; Anterior chamber ; Apoptosis ; Biomedical and Life Sciences ; Biomedicine ; Caspase-3 ; Caspase-9 ; Cell activation ; Cell Biology ; CRISPR ; Eye ; Gadolinium ; Genotype & phenotype ; Glaucoma ; Glial cells ; Glial fibrillary acidic protein ; Immunofluorescence ; Magnetic resonance imaging ; Morphology ; Mutants ; Mutation ; Neurobiology ; Neurosciences ; Optic nerve ; Original Research ; Phenotypes ; Proteins ; Retina ; Retinal ganglion cells ; Transgenic animals ; Transgenic mice ; Visual perception</subject><ispartof>Cellular and molecular neurobiology, 2023-07, Vol.43 (5), p.2021-2033</ispartof><rights>The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2022. Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.</rights><rights>2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c375t-3afb21f4d9f2055914fbfa0cb4f4a7dd179aa2ac804f90e91aafad68bd5d95023</citedby><cites>FETCH-LOGICAL-c375t-3afb21f4d9f2055914fbfa0cb4f4a7dd179aa2ac804f90e91aafad68bd5d95023</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10571-022-01280-x$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10571-022-01280-x$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27923,27924,41487,42556,51318</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36069958$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Cheng, Ying</creatorcontrib><creatorcontrib>Wu, Shen</creatorcontrib><creatorcontrib>Yan, Xuejing</creatorcontrib><creatorcontrib>Liu, Qian</creatorcontrib><creatorcontrib>Lin, Danting</creatorcontrib><creatorcontrib>Zhang, Jingxue</creatorcontrib><creatorcontrib>Wang, Ningli</creatorcontrib><title>Human Pro370Leu Mutant Myocilin Induces the Phenotype of Open-Angle Glaucoma in Transgenic Mice</title><title>Cellular and molecular neurobiology</title><addtitle>Cell Mol Neurobiol</addtitle><addtitle>Cell Mol Neurobiol</addtitle><description>To investigate the characteristics of mutation myocilin proteins and glaucoma pathological phenotype in transgenic mice with full-length human Pro370Leu mutant myocilin gene (Tg-
MYOC
P370L
). Tg-
MYOC
P370L
mice were established using the CRISPR/Cas9 system. Long-term intraocular pressure (IOP) was measured, myocilin protein expressions in anterior chamber angle, retina, optic nerve tissues and aqueous humor were detected by western blot. RBPMS, myocilin, Iba-1 and GFAP expression were visualized by immunofluorescence. H&E staining was applied to assess the ocular angle and retinal morphology. Aqueous humor dynamics were visualized by Gadolinium magnetic resonance imaging (Gd-MRI). TUNEL assay was used to evaluate the specific cell apoptosis in trabecular meshwork and retina. Optomotor and electroretinography tests were employed to evaluate the visual function in Tg-
MYOC
P370L
and wild-type (WT) mice. Homozygous myocilin mutation at position 503 (C > T) was identified by PCR and sequencing in Tg-
MYOC
P370L
mice. Myocilin protein expression was overexpressed in eye tissues of Tg-
MYOC
P370L
mice with reduced myocilin secretion in aqueous humor. H&E staining showed normal histological morphology of anterior chamber angle whereas decreased thickness and nuclei in ganglion cell layer were found (
P
< 0.05). Gd signals were significantly increased in the anterior chamber of Tg-
MYOC
P370L
compared with WT eyes (
P
< 0.05). IOP was elevated in Tg-
MYOC
P370L
mice starting at 5 months of age, with significant RGC loss (
P
< 0.05). Upregulation of caspase-3 and caspase-9 expressions and increased TUNEL-positive cells were found in eyes of Tg-
MYOC
P370L
mice. Excessive activation of retinal glial cells and impaired visual function were detected in Tg-
MYOC
P370L
mice. Tg-
MYOC
P370L
mice can induce the phenotype of open-angle glaucoma, featured as IOP elevation, activated retinal glial cells, loss of RGCs and impaired visual function. These pathologic changes may arise from the abnormal mutant myocilin protein accumulation in the trabecular meshwork and injured aqueous humor drainage. Therefore, Tg-
MYOC
P370L
mice model can serve as an effective animal model for glaucoma research, especially for glaucoma-associated myocilin mutation studies.</description><subject>Animal models</subject><subject>Anterior chamber</subject><subject>Apoptosis</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Caspase-3</subject><subject>Caspase-9</subject><subject>Cell activation</subject><subject>Cell Biology</subject><subject>CRISPR</subject><subject>Eye</subject><subject>Gadolinium</subject><subject>Genotype & phenotype</subject><subject>Glaucoma</subject><subject>Glial cells</subject><subject>Glial fibrillary acidic protein</subject><subject>Immunofluorescence</subject><subject>Magnetic resonance imaging</subject><subject>Morphology</subject><subject>Mutants</subject><subject>Mutation</subject><subject>Neurobiology</subject><subject>Neurosciences</subject><subject>Optic nerve</subject><subject>Original Research</subject><subject>Phenotypes</subject><subject>Proteins</subject><subject>Retina</subject><subject>Retinal ganglion cells</subject><subject>Transgenic animals</subject><subject>Transgenic mice</subject><subject>Visual perception</subject><issn>0272-4340</issn><issn>1573-6830</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><recordid>eNp9kLFuFDEQhi0EIpfAC1AgSzQ0hrG9XttlFIUk0p2SItTG67UvG-3ah72Wcm_Ds_Bk2XABJAqqKeb7_xl9CL2j8IkCyM-FgpCUAGMEKFNAHl6gFRWSk1ZxeIlWwCQjDW_gCB2Xcg8AGkC8Rke8hVZroVbo22WdbMQ3OXEJa1_xps42znizT24Yh4ivYl-dL3i-8_jmzsc073f-548U8PXOR3Iat6PHF6OtLk0WL4HbbGPZ-jg4vBmcf4NeBTsW__Z5nqCvX85vzy7J-vri6ux0TRyXYibcho7R0PQ6MBBC0yZ0wYLrmtBY2fdUamuZdQqaoMFram2wfau6XvRaAOMn6OOhd5fT9-rLbKahOD-ONvpUi2GSUg5SULGgH_5B71PNcfnOMMWUUrptnih2oFxOpWQfzC4Pk817Q8E8-TcH_2bxb375Nw9L6P1zde0m3_-J_Ba-APwAlGUVtz7_vf2f2kcNxpFF</recordid><startdate>20230701</startdate><enddate>20230701</enddate><creator>Cheng, Ying</creator><creator>Wu, Shen</creator><creator>Yan, Xuejing</creator><creator>Liu, Qian</creator><creator>Lin, Danting</creator><creator>Zhang, Jingxue</creator><creator>Wang, Ningli</creator><general>Springer US</general><general>Springer Nature B.V</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20230701</creationdate><title>Human Pro370Leu Mutant Myocilin Induces the Phenotype of Open-Angle Glaucoma in Transgenic Mice</title><author>Cheng, Ying ; Wu, Shen ; Yan, Xuejing ; Liu, Qian ; Lin, Danting ; Zhang, Jingxue ; Wang, Ningli</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c375t-3afb21f4d9f2055914fbfa0cb4f4a7dd179aa2ac804f90e91aafad68bd5d95023</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Animal models</topic><topic>Anterior chamber</topic><topic>Apoptosis</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Caspase-3</topic><topic>Caspase-9</topic><topic>Cell activation</topic><topic>Cell Biology</topic><topic>CRISPR</topic><topic>Eye</topic><topic>Gadolinium</topic><topic>Genotype & phenotype</topic><topic>Glaucoma</topic><topic>Glial cells</topic><topic>Glial fibrillary acidic protein</topic><topic>Immunofluorescence</topic><topic>Magnetic resonance imaging</topic><topic>Morphology</topic><topic>Mutants</topic><topic>Mutation</topic><topic>Neurobiology</topic><topic>Neurosciences</topic><topic>Optic nerve</topic><topic>Original Research</topic><topic>Phenotypes</topic><topic>Proteins</topic><topic>Retina</topic><topic>Retinal ganglion cells</topic><topic>Transgenic animals</topic><topic>Transgenic mice</topic><topic>Visual perception</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Cheng, Ying</creatorcontrib><creatorcontrib>Wu, Shen</creatorcontrib><creatorcontrib>Yan, Xuejing</creatorcontrib><creatorcontrib>Liu, Qian</creatorcontrib><creatorcontrib>Lin, Danting</creatorcontrib><creatorcontrib>Zhang, Jingxue</creatorcontrib><creatorcontrib>Wang, Ningli</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Cellular and molecular neurobiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Cheng, Ying</au><au>Wu, Shen</au><au>Yan, Xuejing</au><au>Liu, Qian</au><au>Lin, Danting</au><au>Zhang, Jingxue</au><au>Wang, Ningli</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Human Pro370Leu Mutant Myocilin Induces the Phenotype of Open-Angle Glaucoma in Transgenic Mice</atitle><jtitle>Cellular and molecular neurobiology</jtitle><stitle>Cell Mol Neurobiol</stitle><addtitle>Cell Mol Neurobiol</addtitle><date>2023-07-01</date><risdate>2023</risdate><volume>43</volume><issue>5</issue><spage>2021</spage><epage>2033</epage><pages>2021-2033</pages><issn>0272-4340</issn><eissn>1573-6830</eissn><abstract>To investigate the characteristics of mutation myocilin proteins and glaucoma pathological phenotype in transgenic mice with full-length human Pro370Leu mutant myocilin gene (Tg-
MYOC
P370L
). Tg-
MYOC
P370L
mice were established using the CRISPR/Cas9 system. Long-term intraocular pressure (IOP) was measured, myocilin protein expressions in anterior chamber angle, retina, optic nerve tissues and aqueous humor were detected by western blot. RBPMS, myocilin, Iba-1 and GFAP expression were visualized by immunofluorescence. H&E staining was applied to assess the ocular angle and retinal morphology. Aqueous humor dynamics were visualized by Gadolinium magnetic resonance imaging (Gd-MRI). TUNEL assay was used to evaluate the specific cell apoptosis in trabecular meshwork and retina. Optomotor and electroretinography tests were employed to evaluate the visual function in Tg-
MYOC
P370L
and wild-type (WT) mice. Homozygous myocilin mutation at position 503 (C > T) was identified by PCR and sequencing in Tg-
MYOC
P370L
mice. Myocilin protein expression was overexpressed in eye tissues of Tg-
MYOC
P370L
mice with reduced myocilin secretion in aqueous humor. H&E staining showed normal histological morphology of anterior chamber angle whereas decreased thickness and nuclei in ganglion cell layer were found (
P
< 0.05). Gd signals were significantly increased in the anterior chamber of Tg-
MYOC
P370L
compared with WT eyes (
P
< 0.05). IOP was elevated in Tg-
MYOC
P370L
mice starting at 5 months of age, with significant RGC loss (
P
< 0.05). Upregulation of caspase-3 and caspase-9 expressions and increased TUNEL-positive cells were found in eyes of Tg-
MYOC
P370L
mice. Excessive activation of retinal glial cells and impaired visual function were detected in Tg-
MYOC
P370L
mice. Tg-
MYOC
P370L
mice can induce the phenotype of open-angle glaucoma, featured as IOP elevation, activated retinal glial cells, loss of RGCs and impaired visual function. These pathologic changes may arise from the abnormal mutant myocilin protein accumulation in the trabecular meshwork and injured aqueous humor drainage. Therefore, Tg-
MYOC
P370L
mice model can serve as an effective animal model for glaucoma research, especially for glaucoma-associated myocilin mutation studies.</abstract><cop>New York</cop><pub>Springer US</pub><pmid>36069958</pmid><doi>10.1007/s10571-022-01280-x</doi><tpages>13</tpages></addata></record> |
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| ispartof | Cellular and molecular neurobiology, 2023-07, Vol.43 (5), p.2021-2033 |
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| source | SpringerLink Journals - AutoHoldings |
| subjects | Animal models Anterior chamber Apoptosis Biomedical and Life Sciences Biomedicine Caspase-3 Caspase-9 Cell activation Cell Biology CRISPR Eye Gadolinium Genotype & phenotype Glaucoma Glial cells Glial fibrillary acidic protein Immunofluorescence Magnetic resonance imaging Morphology Mutants Mutation Neurobiology Neurosciences Optic nerve Original Research Phenotypes Proteins Retina Retinal ganglion cells Transgenic animals Transgenic mice Visual perception |
| title | Human Pro370Leu Mutant Myocilin Induces the Phenotype of Open-Angle Glaucoma in Transgenic Mice |
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