Van't Sant, Lisanne J., White, Joshua J., Hoeijmakers, Jan H. J., Vermeij, Wilbert P. and Jaarsma, Dick ORCID: 0000-0002-0519-2940 (2021). In vivo 5-ethynyluridine (EU) labelling detects reduced transcription in Purkinje cell degeneration mouse mutants, but can itself induce neurodegeneration. Acta Neuropathol. Commun., 9 (1). LONDON: BMC. ISSN 2051-5960

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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.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Van't Sant, Lisanne J.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
White, Joshua J.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Hoeijmakers, Jan H. J.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Vermeij, Wilbert P.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Jaarsma, DickUNSPECIFIEDorcid.org/0000-0002-0519-2940UNSPECIFIED
URN: urn:nbn:de:hbz:38-584103
DOI: 10.1186/s40478-021-01200-y
Journal or Publication Title: Acta Neuropathol. Commun.
Volume: 9
Number: 1
Date: 2021
Publisher: BMC
Place of Publication: LONDON
ISSN: 2051-5960
Language: English
Faculty: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
PRE-MESSENGER-RNA; DNA-DAMAGE; TRANSGENIC MICE; METABOLISM; INHIBITION; NEURONS; STRESS; SCA1; 5-FLUOROURACIL; VISUALIZATIONMultiple languages
NeurosciencesMultiple languages
URI: http://kups.ub.uni-koeln.de/id/eprint/58410

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