ScRNA transcription profile of adult zebrafish podocytes using a novel reporter strain

Cara Brown, Linda J. Mullins, Katrina Wesencraft, Gail McConnell, Mariana Beltran, Neil C. Henderson, Bryan Conway, Scott Hoffmann, Sebastian Rider, John J. Mullins

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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.

Original languageEnglish
Pages (from-to)35-47
Number of pages13
JournalCellular Physiology and Biochemistry
Volume55
Early online date5 May 2021
DOIs
Publication statusE-pub ahead of print - 5 May 2021

Keywords

  • fluorescent reporters
  • podocytes
  • ScRNA-seq
  • zebrafish

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