PUBLICATION
scRNA Transcription Profile of Adult Zebrafish Podocytes Using a Novel Reporter Strain
- Authors
- Brown, C., Mullins, L.J., Wesencraft, K., McConnell, G., Beltran, M., Henderson, N.C., Conway, B., Hoffmann, S., Rider, S., Mullins, J.J.
- ID
- ZDB-PUB-210505-8
- Date
- 2021
- Source
- Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 55: 35-47 (Journal)
- Registered Authors
- Rider, Sebastien
- Keywords
- Zebrafish; scRNA-seq; Podocytes; Fluorescent reporters
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/genetics
- Gene Expression Profiling
- Podocytes/metabolism*
- RNA, Small Cytoplasmic/genetics*
- Transcriptome*
- Zebrafish/genetics*
- PubMed
- 33945241 Full text @ Cell Physiol. Biochem.
Citation
Brown, C., Mullins, L.J., Wesencraft, K., McConnell, G., Beltran, M., Henderson, N.C., Conway, B., Hoffmann, S., Rider, S., Mullins, J.J. (2021) scRNA Transcription Profile of Adult Zebrafish Podocytes Using a Novel Reporter Strain. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology. 55:35-47.
Abstract
Background/aims The role of podocytes is well conserved across species from drosophila to teleosts, and mammals. Identifying the molecular markers that actively maintain the integrity of the podocyte will enable a greater understanding of the changes that lead to damage.
Methods We generated transgenic zebrafish, expressing fluorescent reporters driven by the podocin promoter, for the visualization and isolation of podocytes. We have conducted single cell RNA sequencing (scRNA-seq) on isolated podocytes from a zebrafish reporter line.
Results We demonstrated that the LifeAct-TagRFP-T fluorescent reporter faithfully replicated podocin expression in vivo. We were also able to show spontaneous GCaMP6s fluorescence using light sheet (single plane illumination) microscopy. We identified many podocyte transcripts, encoding proteins related to calcium-binding and actin filament assembly, in common with those expressed in human and mouse mature podocytes.
Conclusion We describe the establishment of novel transgenic zebrafish and their use to identify and isolate podocyte cells for the preparation of a scRNA-seq library from normal podocytes. The scRNA-seq data identifies distinct populations of cells and potential gene switching between clusters. These data provide a foundation for future comparative studies and for exploiting the zebrafish as a model for kidney development, disease, injury and repair.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping