Transcriptomic Profiling of Zebrafish Hair Cells Using RiboTag

Frontiers in Cell and Developmental Biology
Maggie MaternRonna Hertzano

Abstract

The zebrafish inner ear organs and lateral line neuromasts are comprised of a variety of cell types, including mechanosensitive hair cells. Zebrafish hair cells are evolutionarily homologous to mammalian hair cells, and have been particularly useful for studying normal hair cell development and function. However, the relative scarcity of hair cells within these complex organs, as well as the difficulty of fine dissection at early developmental time points, makes hair cell-specific gene expression profiling technically challenging. Cell sorting methods, as well as single-cell RNA-Seq, have proved to be very informative in studying hair cell-specific gene expression. However, these methods require that tissues are dissociated, the processing for which can lead to changes in gene expression prior to RNA extraction. To bypass this problem, we have developed a transgenic zebrafish model to evaluate the translatome of the inner ear and lateral line hair cells in their native tissue environment; the Tg(myo6b:RiboTag) zebrafish. This model expresses both GFP and a hemagglutinin (HA) tagged rpl10a gene under control of the myo6b promoter (myo6b:GFP-2A-rpl10a-3xHA), resulting in HA-tagged ribosomes expressed specifically in hair cells. C...Continue Reading

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Citations

Jun 25, 2020·The Laryngoscope·Ronna HertzanoMaggie S Matern
May 11, 2019·BMC Genomics·María Torres-SánchezDiego San Mauro
Jun 13, 2020·Frontiers in Cellular Neuroscience·Erica SadlerLisa L Cunningham
Sep 28, 2018·Frontiers in Cell and Developmental Biology·Katie S Kindt, Lavinia Sheets
May 6, 2020·Molecular Psychiatry·Michel EngelnMary Kay Lobo
Dec 4, 2020·Environmental Toxicology and Chemistry·Jennifer A FitzgeraldColette Vom Berg

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Datasets Mentioned

BETA
GSE102861
as

Methods Mentioned

BETA
fluorescence
RNA-Seq
scRNA-Seq
pulldown
immunoprecipitation
transgenic
FACS
Flow Cytometry
PCR
pulls down

Software Mentioned

Zebrafish Expression Ontology of Gene Sets ( ZEOGS )
HTSeq
DEseq
TopHat
ZEOGS

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