In vivo Validation of Bimolecular Fluorescence Complementation (BiFC) to Investigate Aggregate Formation in Amyotrophic Lateral Sclerosis (ALS)

• DOI: 10.1007/s12035-020-02238-0
• Publication Date: 2021-01-07T19:12:30Z
• Authors: Emily K. Don; Alina Maschirow; Rowan A. W. Radford; Natalie M. Scherer; Andrés Vidal-Itriago; Alison Hogan; Cindy Maurel; Isabel Formella; Jack J. Stoddart; Thomas E. Hall; Albert Lee; Bingyang Shi; Nicholas J. Cole; Angela S. Laird; Andrew P. Badrock; Roger S. Chung; Marco Morsch
• Keywords: Bimolecular fluorescence complementation

Abstract Amyotrophic lateral sclerosis (ALS) is a form of motor neuron disease (MND) that characterized by the progressive loss neurons within spinal cord, brainstem, and cortex. Although ALS clinically manifests as heterogeneous disease, with varying onset survival, unifying feature presence ubiquitinated cytoplasmic protein inclusion aggregates containing TDP-43. However, precise mechanisms linking inclusions aggregation to neuronal are currently poorly understood. Bimolecular fluorescence complementation (BiFC) takes advantage association fluorophore fragments (non-fluorescent on their own) attached an aggregation-prone interest. Interaction proteins interest allows for fluorescent reporter fold into its native state emit signal. Here, we combined power BiFC advantages zebrafish system validate, optimize, visualize formation ALS-linked in real time vertebrate model. We further provide vivo validation selectivity this technique demonstrate reduced spontaneous self-assembly non-fluorescent introducing mutation. Additionally, report preliminary findings dynamic hallmark Fus TDP-43 corresponding nuclear compartments using BiFC. Overall, our data demonstrates suitability approach study characterize aggregate vivo. Importantly, same principle can be applied context other neurodegenerative diseases has therefore critical implications advance understanding pathologies underlie aberrant aggregation.