In vivo 5-ethynyluridine (EU) labelling detects reduced transcription in Purkinje cell degeneration mouse mutants, but can itself induce neurodegeneration

Lisanne J. van’t Sant, Joshua J. White, Jan H.J. Hoeijmakers, Wilbert P. Vermeij, Dick Jaarsma*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

6 Citations (Scopus)

Abstract

Fluorescent staining of newly transcribed RNA via metabolic labelling with 5-ethynyluridine (EU) and click chemistry enables visualisation of changes in transcription, such as in conditions of cellular stress. Here, we tested whether EU labelling can be used to examine transcription in vivo in mouse models of nervous system disorders. We show that injection of EU directly into the cerebellum results in reproducible labelling of newly transcribed RNA in cerebellar neurons and glia, with cell type-specific differences in relative labelling intensities, such as Purkinje cells exhibiting the highest levels. We also observed EU-labelling accumulating into cytoplasmic inclusions, indicating that EU, like other modified uridines, may introduce non-physiological properties in labelled RNAs. Additionally, we found that EU induces Purkinje cell degeneration nine days after EU injection, suggesting that EU incorporation not only results in abnormal RNA transcripts, but also eventually becomes neurotoxic in highly transcriptionally-active neurons. However, short post-injection intervals of EU labelling in both a Purkinje cell-specific DNA repair-deficient mouse model and a mouse model of spinocerebellar ataxia 1 revealed reduced transcription in Purkinje cells compared to controls. We combined EU labelling with immunohistology to correlate altered EU staining with pathological markers, such as genotoxic signalling factors. These data indicate that the EU-labelling method provided here can be used to identify changes in transcription in vivo in nervous system disease models.

Original languageEnglish
Article number94
JournalActa neuropathologica communications
Volume9
Issue number1
DOIs
Publication statusPublished - 21 May 2021

Bibliographical note

Funding Information:
We are grateful to Elise D. Haasdijk for histological work; Francois Blot and Renata M.C. Brandt for assistance with mouse experiments; María B. Birkisdóttir for valuable discussions; Harry T. Orr for providing ATXN1[82Q] transgenic mice. We acknowledge financial support from ZonMW Memorabel (733050810) and NWO Building Blocks of Life. Support from Oncode (Dutch Cancer Institute) and EJP-RD TC-NER RD20-113 is acknowledged by J.H.J.H. and W.P.V.. J.H.J.H. is also supported by the ERC Advanced Grants DamAge and Dam2Age, NIH grant (PO1 AG017242) and the Deutsche Forschungsgemeinschaft (SFB 829). J.J.W is funded by NWO-Veni.

Funding Information:
We are grateful to Elise D. Haasdijk for histological work; Francois Blot and Renata M.C. Brandt for assistance with mouse experiments; Mar?a B. Birkisd?ttir for valuable discussions; Harry T. Orr for providing ATXN1[82Q] transgenic mice. We acknowledge financial support from ZonMW Memorabel (733050810) and NWO Building Blocks of Life. Support from Oncode (Dutch Cancer Institute) and EJP-RD TC-NER RD20-113 is acknowledged by J.H.J.H. and W.P.V. J.H.J.H. is also supported by the ERC Advanced Grants DamAge and Dam2Age, NIH grant (PO1 AG017242) and the Deutsche Forschungsgemeinschaft (SFB 829). J.J.W is funded by NWO-Veni.

Publisher Copyright:
© 2021, The Author(s).

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