Visualize neuronal membrane cholesterol with split-fluorescent protein tagged YDQA sensor

TL;DR

This study creates a new sensor to visualize cholesterol in neurons, revealing abnormal cholesterol patterns in Alzheimer's disease models.

Contribution

A novel sfPMcho sensor combining split-fluorescent protein tags and YDQA for in vivo neuronal membrane cholesterol detection.

Findings

sfPMcho efficiently detects neuronal plasma membrane cholesterol in cell and C. elegans models.

In AD-related mouse models, neuronal cholesterol becomes sparse in bodies but accumulates in nerve fibers.

The sensor reveals cellular-level cholesterol abnormalities in Alzheimer's disease pathology.

Abstract

Cholesterol is a major component of the cellular plasma membrane (PM), and its homeostasis is essential for brain health. Dysregulated cholesterol homeostasis has been strongly implicated in the pathogenesis of various neurological disorders, including Alzheimer’s disease (AD). However, in vivo visualization of cholesterol has remained challenging, hindering a comprehensive understanding of AD pathology. In this study, we generated a new sensor combining the split-fluorescent protein tags with YDQA, a derivate of cholesterol-dependent cytolysin PFO. Through a series of validations in cell and C. elegans models, we demonstrate that the new sensor (name as sfPMcho) efficiently detects neuronal PM cholesterol. We further applied this sensor in 5X FAD and APOE KO mice models and revealed the cholesterol changes within neurons. PM cholesterol became sparse and locally aggregated in neuron bodies but significantly accumulated in nerve fibers. Collectively, this study provides a new tool for detecting neuronal PM cholesterol in vivo and uncovers cholesterol abnormalities in AD-related pathology at the cellular level. Further development based on this sensor or a similar strategy is to be expected.

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