PUBLICATION
Spatial detection of mitochondrial DNA and RNA in tissues
- Authors
- Giarmarco, M., Seto, J., Brock, D., Brockerhoff, S.
- ID
- ZDB-PUB-240529-12
- Date
- 2024
- Source
- Frontiers in cell and developmental biology 12: 13467781346778 (Journal)
- Registered Authors
- Brockerhoff, Susan
- Keywords
- RNAscope, mitochondria, mtDNA, mtRNA, multiplex imaging, photoreceptor, spatial biology, zebrafish
- MeSH Terms
- none
- PubMed
- 38808224 Full text @ Front Cell Dev Biol
Citation
Giarmarco, M., Seto, J., Brock, D., Brockerhoff, S. (2024) Spatial detection of mitochondrial DNA and RNA in tissues. Frontiers in cell and developmental biology. 12:13467781346778.
Abstract
Background Mitochondrial health has gained attention in a number of diseases, both as an indicator of disease state and as a potential therapeutic target. The quality and amount of mitochondrial DNA (mtDNA) and RNA (mtRNA) can be important indicators of mitochondrial and cell health, but are difficult to measure in complex tissues.
Methods mtDNA and mtRNA in zebrafish retina samples were fluorescently labeled using RNAscope™ in situ hybridization, then mitochondria were stained using immunohistochemistry. Pretreatment with RNase was used for validation. Confocal images were collected and analyzed, and relative amounts of mtDNA and mtRNA were reported. Findings regarding mtDNA were confirmed using qPCR.
Results Signals from probes detecting mtDNA and mtRNA were localized to mitochondria, and were differentially sensitive to RNase. This labeling strategy allows for quantification of relative mtDNA and mtRNA levels in individual cells. As a demonstration of the method in a complex tissue, single photoreceptors in zebrafish retina were analyzed for mtDNA and mtRNA content. An increase in mtRNA but not mtDNA coincides with proliferation of mitochondria at night in cones. A similar trend was measured in rods.
Discussion Mitochondrial gene expression is an important component of cell adaptations to disease, stress, or aging. This method enables the study of mtDNA and mtRNA in single cells of an intact, complex tissue. The protocol presented here uses commercially-available tools, and is adaptable to a range of species and tissue types.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping