An Ex Vivo Study on Release, Uptake, and miRNA Profile of Exosomes in Rat Lens
Purpose. To identify the ability of releasing and uptaking exosomes in rat lens and characterize the exosomal microRNA profile of lens-derived exosomes. Methods. The rat lenses were cultured ex vivo and the medium was collected. The exosomes were isolated from medium and measured in size and concent...
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| Format: | Article |
| Language: | English |
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Wiley
2022-01-01
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| Series: | Journal of Ophthalmology |
| Online Access: | http://dx.doi.org/10.1155/2022/6706172 |
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| author | Junfang Zhang Jiawei Luo Guowei Zhang Bai Qin Xiumei Ren Huaijin Guan |
| author_facet | Junfang Zhang Jiawei Luo Guowei Zhang Bai Qin Xiumei Ren Huaijin Guan |
| author_sort | Junfang Zhang |
| collection | DOAJ |
| description | Purpose. To identify the ability of releasing and uptaking exosomes in rat lens and characterize the exosomal microRNA profile of lens-derived exosomes. Methods. The rat lenses were cultured ex vivo and the medium was collected. The exosomes were isolated from medium and measured in size and concentration by nanoflow cytometry (nFCM) and transmission electron microscopy (TEM) and verified with CD63 and TSG101 by Western blot. The miRNAs in exosomes released from lens epithelial cells (LECs) were sequenced. The plasma exosomes labeled by PKH26 were used to verify the exosomes uptake LECs, and their colocalized fluorescence was imaged by confocal microscopy. Results. LECs released numerous exosomes into the medium through the capsule, which contained abundant miRNAs. The most abundant miRNAs included miR-184, let-7c-5p, let-7a-5p, let-7b-5p, let-7f-5p, miR-125a-5p, miR-204-5p, miR-125b-5p, miR-1b, and miR-23a-3p. The LECs but not the lens fibre cells showed exosome uptake. The LECs uptake more PKH26-labeled exosomes at day 7 than day 3 and day 14. Conclusions. Our results suggested that LECs can release and uptake exosomes through the capsule. Exosomes may be an important way for the lens to communicate among LECs, aqueous humour, vitreous body, and other ocular tissues. |
| format | Article |
| id | doaj-art-3c0a446a6d7a479e922cd4d684af5233 |
| institution | Kabale University |
| issn | 2090-0058 |
| language | English |
| publishDate | 2022-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Ophthalmology |
| spelling | doaj-art-3c0a446a6d7a479e922cd4d684af52332025-02-03T01:22:44ZengWileyJournal of Ophthalmology2090-00582022-01-01202210.1155/2022/6706172An Ex Vivo Study on Release, Uptake, and miRNA Profile of Exosomes in Rat LensJunfang Zhang0Jiawei Luo1Guowei Zhang2Bai Qin3Xiumei Ren4Huaijin Guan5Eye InstituteEye InstituteEye InstituteEye InstituteShanghai Municipal Hospital of Traditional Chinese MedicineEye InstitutePurpose. To identify the ability of releasing and uptaking exosomes in rat lens and characterize the exosomal microRNA profile of lens-derived exosomes. Methods. The rat lenses were cultured ex vivo and the medium was collected. The exosomes were isolated from medium and measured in size and concentration by nanoflow cytometry (nFCM) and transmission electron microscopy (TEM) and verified with CD63 and TSG101 by Western blot. The miRNAs in exosomes released from lens epithelial cells (LECs) were sequenced. The plasma exosomes labeled by PKH26 were used to verify the exosomes uptake LECs, and their colocalized fluorescence was imaged by confocal microscopy. Results. LECs released numerous exosomes into the medium through the capsule, which contained abundant miRNAs. The most abundant miRNAs included miR-184, let-7c-5p, let-7a-5p, let-7b-5p, let-7f-5p, miR-125a-5p, miR-204-5p, miR-125b-5p, miR-1b, and miR-23a-3p. The LECs but not the lens fibre cells showed exosome uptake. The LECs uptake more PKH26-labeled exosomes at day 7 than day 3 and day 14. Conclusions. Our results suggested that LECs can release and uptake exosomes through the capsule. Exosomes may be an important way for the lens to communicate among LECs, aqueous humour, vitreous body, and other ocular tissues.http://dx.doi.org/10.1155/2022/6706172 |
| spellingShingle | Junfang Zhang Jiawei Luo Guowei Zhang Bai Qin Xiumei Ren Huaijin Guan An Ex Vivo Study on Release, Uptake, and miRNA Profile of Exosomes in Rat Lens Journal of Ophthalmology |
| title | An Ex Vivo Study on Release, Uptake, and miRNA Profile of Exosomes in Rat Lens |
| title_full | An Ex Vivo Study on Release, Uptake, and miRNA Profile of Exosomes in Rat Lens |
| title_fullStr | An Ex Vivo Study on Release, Uptake, and miRNA Profile of Exosomes in Rat Lens |
| title_full_unstemmed | An Ex Vivo Study on Release, Uptake, and miRNA Profile of Exosomes in Rat Lens |
| title_short | An Ex Vivo Study on Release, Uptake, and miRNA Profile of Exosomes in Rat Lens |
| title_sort | ex vivo study on release uptake and mirna profile of exosomes in rat lens |
| url | http://dx.doi.org/10.1155/2022/6706172 |
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