βB2-Crystallin Promotes Axonal Regeneration in the Injured Optic Nerve in Adult Rats
The purpose of the study was to further scrutinize the potential of βB2-crystallin in supporting regeneration of injured retinal ganglion cell axons both in vitro and in vivo. Retinal explants obtained from animals after treatment either with lens injury (LI) alone or with combined LI 5 days or 3 da...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2015-09-01
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| Series: | Cell Transplantation |
| Online Access: | https://doi.org/10.3727/096368914X684583 |
| _version_ | 1818777469874864128 |
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| author | Michael R. R. Böhm Verena Prokosch Matthias Brückner Sarah Pfrommer Harutyun Melkonyan Solon Thanos |
| author_facet | Michael R. R. Böhm Verena Prokosch Matthias Brückner Sarah Pfrommer Harutyun Melkonyan Solon Thanos |
| author_sort | Michael R. R. Böhm |
| collection | DOAJ |
| description | The purpose of the study was to further scrutinize the potential of βB2-crystallin in supporting regeneration of injured retinal ganglion cell axons both in vitro and in vivo. Retinal explants obtained from animals after treatment either with lens injury (LI) alone or with combined LI 5 days or 3 days before or simultaneously with an optic nerve crush (ONC) were cultured for 96 h under regenerative conditions, and the regenerating axons were quantified and compared with untreated controls. These measurements were then repeated with LI replaced by intravitreal injections of γ-crystallin and β-crystallin at 5 days before ONC. Finally, bB2-crystallin-overexpressing transfected neural progenitor cells (bB2-crystallin-NPCs) in the eye were studied after crushing the optic nerve in vivo. Regeneration was monitored with the aid of immunoblotting of the retina and optic nerve both distal and proximal to the lesion site, and this was compared with controls that received injections of phosphate buffer only. LI performed 5 days or 3 days before ONC significantly promoted axonal outgrowth in vitro ( p < 0.001), while LI performed alone before explantation did not. Intravitreal injections of β-crystallin and γ-crystallin mimicked the effects of LI and significantly increased axonal regeneration in culture at the same time intervals ( p < 0.001). Western blot analysis revealed that crystallins were present in the proximal optic nerve stump at the lesion site in ONC, but were neither expressed in the undamaged distal optic nerve nor in uninjured tissue. bB2-crystallin-NPCs supported the regeneration of cut optic nerve axons within the distal optic nerve stump in vivo. The reported data suggest that bB2-crystallin-producing “cell factories” could be used to provide novel therapeutic drugs for central nervous system injuries. |
| first_indexed | 2024-12-18T11:29:20Z |
| format | Article |
| id | doaj.art-515e22c0ca094723b10022d28f994df1 |
| institution | Directory Open Access Journal |
| issn | 0963-6897 1555-3892 |
| language | English |
| last_indexed | 2024-12-18T11:29:20Z |
| publishDate | 2015-09-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Cell Transplantation |
| spelling | doaj.art-515e22c0ca094723b10022d28f994df12022-12-21T21:09:37ZengSAGE PublishingCell Transplantation0963-68971555-38922015-09-012410.3727/096368914X684583βB2-Crystallin Promotes Axonal Regeneration in the Injured Optic Nerve in Adult RatsMichael R. R. Böhm0Verena Prokosch1Matthias Brückner2Sarah Pfrommer3Harutyun Melkonyan4Solon Thanos5 Interdisciplinary Center for Clinical Research, School of Medicine, University of Münster, Albert-Schweitzer-Campus 1, Münster, Germany Institute for Experimental Ophthalmology, School of Medicine, University of Münster, Albert-Schweitzer-Campus 1, Münster, Germany Institute for Experimental Ophthalmology, School of Medicine, University of Münster, Albert-Schweitzer-Campus 1, Münster, Germany Institute for Experimental Ophthalmology, School of Medicine, University of Münster, Albert-Schweitzer-Campus 1, Münster, Germany Institute for Experimental Ophthalmology, School of Medicine, University of Münster, Albert-Schweitzer-Campus 1, Münster, Germany Interdisciplinary Center for Clinical Research, School of Medicine, University of Münster, Albert-Schweitzer-Campus 1, Münster, GermanyThe purpose of the study was to further scrutinize the potential of βB2-crystallin in supporting regeneration of injured retinal ganglion cell axons both in vitro and in vivo. Retinal explants obtained from animals after treatment either with lens injury (LI) alone or with combined LI 5 days or 3 days before or simultaneously with an optic nerve crush (ONC) were cultured for 96 h under regenerative conditions, and the regenerating axons were quantified and compared with untreated controls. These measurements were then repeated with LI replaced by intravitreal injections of γ-crystallin and β-crystallin at 5 days before ONC. Finally, bB2-crystallin-overexpressing transfected neural progenitor cells (bB2-crystallin-NPCs) in the eye were studied after crushing the optic nerve in vivo. Regeneration was monitored with the aid of immunoblotting of the retina and optic nerve both distal and proximal to the lesion site, and this was compared with controls that received injections of phosphate buffer only. LI performed 5 days or 3 days before ONC significantly promoted axonal outgrowth in vitro ( p < 0.001), while LI performed alone before explantation did not. Intravitreal injections of β-crystallin and γ-crystallin mimicked the effects of LI and significantly increased axonal regeneration in culture at the same time intervals ( p < 0.001). Western blot analysis revealed that crystallins were present in the proximal optic nerve stump at the lesion site in ONC, but were neither expressed in the undamaged distal optic nerve nor in uninjured tissue. bB2-crystallin-NPCs supported the regeneration of cut optic nerve axons within the distal optic nerve stump in vivo. The reported data suggest that bB2-crystallin-producing “cell factories” could be used to provide novel therapeutic drugs for central nervous system injuries.https://doi.org/10.3727/096368914X684583 |
| spellingShingle | Michael R. R. Böhm Verena Prokosch Matthias Brückner Sarah Pfrommer Harutyun Melkonyan Solon Thanos βB2-Crystallin Promotes Axonal Regeneration in the Injured Optic Nerve in Adult Rats Cell Transplantation |
| title | βB2-Crystallin Promotes Axonal Regeneration in the Injured Optic Nerve in Adult Rats |
| title_full | βB2-Crystallin Promotes Axonal Regeneration in the Injured Optic Nerve in Adult Rats |
| title_fullStr | βB2-Crystallin Promotes Axonal Regeneration in the Injured Optic Nerve in Adult Rats |
| title_full_unstemmed | βB2-Crystallin Promotes Axonal Regeneration in the Injured Optic Nerve in Adult Rats |
| title_short | βB2-Crystallin Promotes Axonal Regeneration in the Injured Optic Nerve in Adult Rats |
| title_sort | βb2 crystallin promotes axonal regeneration in the injured optic nerve in adult rats |
| url | https://doi.org/10.3727/096368914X684583 |
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