Comparison of Retinal Metabolic Activity and Structural Development between rd10 Mice and Normal Mice Using Multiphoton Fluorescence Lifetime Imaging Microscopy
Fluorescence lifetime imaging microscopy (FLIM) is a technique that analyzes the metabolic state of tissues based on the spatial distribution of fluorescence lifetimes of certain interacting molecules. We used multiphoton FLIM to study the metabolic state of developing C57BL6/J and rd10 retinas base...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-01-01
|
| Series: | Current Issues in Molecular Biology |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1467-3045/46/1/39 |
| _version_ | 1827372528950050816 |
|---|---|
| author | Erin Su Niranjana Kesavamoorthy Jason A. Junge Mengmei Zheng Cheryl Mae Craft Hossein Ameri |
| author_facet | Erin Su Niranjana Kesavamoorthy Jason A. Junge Mengmei Zheng Cheryl Mae Craft Hossein Ameri |
| author_sort | Erin Su |
| collection | DOAJ |
| description | Fluorescence lifetime imaging microscopy (FLIM) is a technique that analyzes the metabolic state of tissues based on the spatial distribution of fluorescence lifetimes of certain interacting molecules. We used multiphoton FLIM to study the metabolic state of developing C57BL6/J and rd10 retinas based on the fluorescence lifetimes of free versus bound nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate (NAD(P)H), with free NAD(P)H percentages suggesting increased glycolysis and bound NAD(P)H percentages indicating oxidative phosphorylation. The mice were sacrificed and enucleated at various time points throughout their first 3 months of life. The isolated eyecups were fixed, sectioned using a polyacrylamide gel embedding technique, and then analyzed with FLIM. The results suggested that in both C57BL6/J mice and rd10 mice, oxidative phosphorylation initially decreased and then increased, plateauing over time. This trend, however, was accelerated in rd10 mice, with its turning point occurring at p10 versus the p30 turning point in C57BL6/J mice. There was also a noticeable difference in oxidative phosphorylation rates between the outer and inner retinas in both strains, with greater oxidative phosphorylation present in the latter. A greater understanding of rd10 and WT metabolic changes during retinal development may provide deeper insights into retinal degeneration and facilitate the development of future treatments. |
| first_indexed | 2024-03-08T11:01:01Z |
| format | Article |
| id | doaj.art-444080ea954d41c8950bcd5f290e1061 |
| institution | Directory Open Access Journal |
| issn | 1467-3037 1467-3045 |
| language | English |
| last_indexed | 2024-03-08T11:01:01Z |
| publishDate | 2024-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Current Issues in Molecular Biology |
| spelling | doaj.art-444080ea954d41c8950bcd5f290e10612024-01-26T15:50:50ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452024-01-0146161262010.3390/cimb46010039Comparison of Retinal Metabolic Activity and Structural Development between rd10 Mice and Normal Mice Using Multiphoton Fluorescence Lifetime Imaging MicroscopyErin Su0Niranjana Kesavamoorthy1Jason A. Junge2Mengmei Zheng3Cheryl Mae Craft4Hossein Ameri5Department of Ophthalmology, USC Roski Eye Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USADepartment of Ophthalmology, USC Roski Eye Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USADepartment of Biological Sciences, David Dornsife College of Letters Arts and Sciences, University of Southern California Dana, Los Angeles, CA 90089, USADepartment of Ophthalmology, USC Roski Eye Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USADepartment of Ophthalmology, USC Roski Eye Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USADepartment of Ophthalmology, USC Roski Eye Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USAFluorescence lifetime imaging microscopy (FLIM) is a technique that analyzes the metabolic state of tissues based on the spatial distribution of fluorescence lifetimes of certain interacting molecules. We used multiphoton FLIM to study the metabolic state of developing C57BL6/J and rd10 retinas based on the fluorescence lifetimes of free versus bound nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate (NAD(P)H), with free NAD(P)H percentages suggesting increased glycolysis and bound NAD(P)H percentages indicating oxidative phosphorylation. The mice were sacrificed and enucleated at various time points throughout their first 3 months of life. The isolated eyecups were fixed, sectioned using a polyacrylamide gel embedding technique, and then analyzed with FLIM. The results suggested that in both C57BL6/J mice and rd10 mice, oxidative phosphorylation initially decreased and then increased, plateauing over time. This trend, however, was accelerated in rd10 mice, with its turning point occurring at p10 versus the p30 turning point in C57BL6/J mice. There was also a noticeable difference in oxidative phosphorylation rates between the outer and inner retinas in both strains, with greater oxidative phosphorylation present in the latter. A greater understanding of rd10 and WT metabolic changes during retinal development may provide deeper insights into retinal degeneration and facilitate the development of future treatments.https://www.mdpi.com/1467-3045/46/1/39retinamultiphoton fluorescence lifetime imaging microscopyFLIMNAD(P)Hglycolysisoxidative phosphorylation |
| spellingShingle | Erin Su Niranjana Kesavamoorthy Jason A. Junge Mengmei Zheng Cheryl Mae Craft Hossein Ameri Comparison of Retinal Metabolic Activity and Structural Development between rd10 Mice and Normal Mice Using Multiphoton Fluorescence Lifetime Imaging Microscopy Current Issues in Molecular Biology retina multiphoton fluorescence lifetime imaging microscopy FLIM NAD(P)H glycolysis oxidative phosphorylation |
| title | Comparison of Retinal Metabolic Activity and Structural Development between rd10 Mice and Normal Mice Using Multiphoton Fluorescence Lifetime Imaging Microscopy |
| title_full | Comparison of Retinal Metabolic Activity and Structural Development between rd10 Mice and Normal Mice Using Multiphoton Fluorescence Lifetime Imaging Microscopy |
| title_fullStr | Comparison of Retinal Metabolic Activity and Structural Development between rd10 Mice and Normal Mice Using Multiphoton Fluorescence Lifetime Imaging Microscopy |
| title_full_unstemmed | Comparison of Retinal Metabolic Activity and Structural Development between rd10 Mice and Normal Mice Using Multiphoton Fluorescence Lifetime Imaging Microscopy |
| title_short | Comparison of Retinal Metabolic Activity and Structural Development between rd10 Mice and Normal Mice Using Multiphoton Fluorescence Lifetime Imaging Microscopy |
| title_sort | comparison of retinal metabolic activity and structural development between rd10 mice and normal mice using multiphoton fluorescence lifetime imaging microscopy |
| topic | retina multiphoton fluorescence lifetime imaging microscopy FLIM NAD(P)H glycolysis oxidative phosphorylation |
| url | https://www.mdpi.com/1467-3045/46/1/39 |
| work_keys_str_mv | AT erinsu comparisonofretinalmetabolicactivityandstructuraldevelopmentbetweenrd10miceandnormalmiceusingmultiphotonfluorescencelifetimeimagingmicroscopy AT niranjanakesavamoorthy comparisonofretinalmetabolicactivityandstructuraldevelopmentbetweenrd10miceandnormalmiceusingmultiphotonfluorescencelifetimeimagingmicroscopy AT jasonajunge comparisonofretinalmetabolicactivityandstructuraldevelopmentbetweenrd10miceandnormalmiceusingmultiphotonfluorescencelifetimeimagingmicroscopy AT mengmeizheng comparisonofretinalmetabolicactivityandstructuraldevelopmentbetweenrd10miceandnormalmiceusingmultiphotonfluorescencelifetimeimagingmicroscopy AT cherylmaecraft comparisonofretinalmetabolicactivityandstructuraldevelopmentbetweenrd10miceandnormalmiceusingmultiphotonfluorescencelifetimeimagingmicroscopy AT hosseinameri comparisonofretinalmetabolicactivityandstructuraldevelopmentbetweenrd10miceandnormalmiceusingmultiphotonfluorescencelifetimeimagingmicroscopy |