Selection of red fluorescent protein for genetic labeling of mitochondria and intercellular transfer of viable mitochondria
Abstract The phenomenon of intercellular mitochondrial transfer has attracted great attention in various fields of research, including stem cell biology. Elucidating the mechanism of mitochondrial transfer from healthy stem cells to cells with mitochondrial dysfunction may lead to the development of...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-11-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-24297-0 |
| _version_ | 1811309224887582720 |
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| author | Isamu Taiko Chika Takano Masayuki Nomoto Shingo Hayashida Kazunori Kanemaru Toshio Miki |
| author_facet | Isamu Taiko Chika Takano Masayuki Nomoto Shingo Hayashida Kazunori Kanemaru Toshio Miki |
| author_sort | Isamu Taiko |
| collection | DOAJ |
| description | Abstract The phenomenon of intercellular mitochondrial transfer has attracted great attention in various fields of research, including stem cell biology. Elucidating the mechanism of mitochondrial transfer from healthy stem cells to cells with mitochondrial dysfunction may lead to the development of novel stem cell therapies to treat mitochondrial diseases, among other advances. To visually evaluate and analyze the mitochondrial transfer process, dual fluorescent labeling systems are often used to distinguish the mitochondria of donor and recipient cells. Although enhanced green fluorescent protein (EGFP) has been well-characterized for labeling mitochondria, other colors of fluorescent protein have been less extensively evaluated in the context of mitochondrial transfer. Here, we generated different lentiviral vectors with mitochondria-targeted red fluorescent proteins (RFPs), including DsRed, mCherry (both from Discosoma sp.) Kusabira orange (mKOκ, from Verrillofungia concinna), and TurboRFP (from Entacmaea quadricolor). Among these proteins, mitochondria-targeted DsRed and its variant mCherry often generated bright aggregates in the lysosome while other proteins did not. We further validated that TurboRFP-labeled mitochondria were successfully transferred from amniotic epithelial cells, one of the candidates for donor stem cells, to mitochondria-damaged recipient cells without losing the membrane potential. Our study provides new insight into the genetic labeling of mitochondria with red fluorescent proteins, which may be utilized to analyze the mechanism of intercellular mitochondrial transfer. |
| first_indexed | 2024-04-13T09:38:30Z |
| format | Article |
| id | doaj.art-0f0e13b9c1cf441d9f1f77ebb05f2879 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-13T09:38:30Z |
| publishDate | 2022-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-0f0e13b9c1cf441d9f1f77ebb05f28792022-12-22T02:52:00ZengNature PortfolioScientific Reports2045-23222022-11-0112111010.1038/s41598-022-24297-0Selection of red fluorescent protein for genetic labeling of mitochondria and intercellular transfer of viable mitochondriaIsamu Taiko0Chika Takano1Masayuki Nomoto2Shingo Hayashida3Kazunori Kanemaru4Toshio Miki5Department of Physiology, Nihon University School of MedicineDivision of Microbiology, Department of Pathology and Microbiology, Nihon University School of MedicineDivision of Respiratory Medicine, Department of Internal Medicine, Nihon University School of MedicineDepartment of Pediatrics and Child Health, Nihon University School of MedicineDepartment of Physiology, Nihon University School of MedicineDepartment of Physiology, Nihon University School of MedicineAbstract The phenomenon of intercellular mitochondrial transfer has attracted great attention in various fields of research, including stem cell biology. Elucidating the mechanism of mitochondrial transfer from healthy stem cells to cells with mitochondrial dysfunction may lead to the development of novel stem cell therapies to treat mitochondrial diseases, among other advances. To visually evaluate and analyze the mitochondrial transfer process, dual fluorescent labeling systems are often used to distinguish the mitochondria of donor and recipient cells. Although enhanced green fluorescent protein (EGFP) has been well-characterized for labeling mitochondria, other colors of fluorescent protein have been less extensively evaluated in the context of mitochondrial transfer. Here, we generated different lentiviral vectors with mitochondria-targeted red fluorescent proteins (RFPs), including DsRed, mCherry (both from Discosoma sp.) Kusabira orange (mKOκ, from Verrillofungia concinna), and TurboRFP (from Entacmaea quadricolor). Among these proteins, mitochondria-targeted DsRed and its variant mCherry often generated bright aggregates in the lysosome while other proteins did not. We further validated that TurboRFP-labeled mitochondria were successfully transferred from amniotic epithelial cells, one of the candidates for donor stem cells, to mitochondria-damaged recipient cells without losing the membrane potential. Our study provides new insight into the genetic labeling of mitochondria with red fluorescent proteins, which may be utilized to analyze the mechanism of intercellular mitochondrial transfer.https://doi.org/10.1038/s41598-022-24297-0 |
| spellingShingle | Isamu Taiko Chika Takano Masayuki Nomoto Shingo Hayashida Kazunori Kanemaru Toshio Miki Selection of red fluorescent protein for genetic labeling of mitochondria and intercellular transfer of viable mitochondria Scientific Reports |
| title | Selection of red fluorescent protein for genetic labeling of mitochondria and intercellular transfer of viable mitochondria |
| title_full | Selection of red fluorescent protein for genetic labeling of mitochondria and intercellular transfer of viable mitochondria |
| title_fullStr | Selection of red fluorescent protein for genetic labeling of mitochondria and intercellular transfer of viable mitochondria |
| title_full_unstemmed | Selection of red fluorescent protein for genetic labeling of mitochondria and intercellular transfer of viable mitochondria |
| title_short | Selection of red fluorescent protein for genetic labeling of mitochondria and intercellular transfer of viable mitochondria |
| title_sort | selection of red fluorescent protein for genetic labeling of mitochondria and intercellular transfer of viable mitochondria |
| url | https://doi.org/10.1038/s41598-022-24297-0 |
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