Recent Microscopy Advances and the Applications to Huntington’s Disease Research

Abstract

Huntingtin is a 3144 amino acid protein defined as a scaffold protein with many intracellular locations that suggest functions in these compartments. Expansion of the CAG DNA tract in the huntingtin first exon is the cause of Huntington’s disease. An important tool in understanding the biological functions of huntingtin is molecular imaging at the single-cell level by microscopy and nanoscopy. The evolution of these technologies has accelerated since the Nobel Prize in Chemistry was awarded in 2014 for super-resolution nanoscopy. We are in a new era of light imaging at the single-cell level, not just for protein location, but also for protein conformation and biochemical function. Large-scale microscopy-based screening is also being accelerated by a coincident development of machine-based learning that offers a framework for truly unbiased data acquisition and analysis at very large scales. This review will summarize the newest technologies in light, electron, and atomic force microscopy in the context of unique challenges with huntingtin cell biology and biochemistry.

Accelerated protein retention expansion microscopy using microwave radiation

“…al 2021, Campbell et al 2021, Katoh 2016, Martinez et al 2020, Pernal et al 2020, Vanheusden et al 2020).3.4 M/W ExM maintains the superior resolution of ExM to resolve synaptic components ExM has been demonstrated to yield superior resolution for subcellular structures in neural tissue and has since become a vital tool in neuroscience(Babi et al 2022, Eichler et al 2017, Freifeld et al 2017, Gallagher & Zhao 2021, Karagiannis & Boyden 2018, Sneve & Piatkevich 2021. To demonstrate that M/W ExM retains the resolution for subcellular structures in neurons at the synaptic level as had been shown with conventional ExM, we dual-labeled free-floating vibratome tectal sections with antibodies against PSD-95 (to label excitatory synapses, Fig.5A-B) and β-tubulin (to label the neuronal processes Fig.5C-D).…”