In Vivo Bioluminescence Imaging of the Intracerebral Fibroin-controlled AAV-α-synuclein Diffusion for Monitoring the Central Nervous System and Peripheral Expression

Overview

Journal Sci Rep

Specialty Science

Date 2024 Apr 27

PMID 38678103

Authors

Claire Mazzocco

Coralie Genevois

Qin Li

Evelyne Doudnikoff

Nathalie Dutheil

Thierry Leste-Lasserre

Marie-Laure Arotcarena

Erwan Bezard

Affiliations

Soon will be listed here.

Abstract

Among the several animal models of α-synucleinopathies, the well-known viral vector-mediated delivery of wild-type or mutated (A53T) α-synuclein requires new tools to increase the lesion in mice and follow up in vivo expression. To this end, we developed a bioluminescent expression reporter of the human A53T-α-synuclein gene using the NanoLuc system into an AAV2/9, embedded or not in a fibroin solution to stabilise its expression in space and time. We first verified the expression of the fused protein in vitro on transfected cells by bioluminescence and Western blotting. Next, two groups of C57Bl6Jr mice were unilaterally injected with the AAV-NanoLuc-human-A53T-α-synuclein above the substantia nigra combined (or not) with fibroin. We first show that the in vivo cerebral bioluminescence signal was more intense in the presence of fibroin. Using immunohistochemistry, we find that the human-A53T-α-synuclein protein is more restricted to the ipsilateral side with an overall greater magnitude of the lesion when fibroin was added. However, we also detected a bioluminescence signal in peripheral organs in both conditions, confirmed by the presence of viral DNA corresponding to the injected AAV in the liver using qPCR.

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