Spatiotemporal Imaging of Zinc Ions in Zebrafish Live Brain Tissue Enabled by Fluorescent Bionanoprobes
Abstrakt
The zebrafish is a powerful model organism to study the mechanisms governing transition metal ions within whole brain tissue. Zinc is one of the most abundant metal ions in the brain, playing a critical pathophysiological role in neurodegenerative diseases.
The homeostasis of free, ionic zinc (Zn(2+)) is a key intersection point in many of these diseases, including Alzheimer's disease and Parkinson's disease. A Zn(2+) imbalance can eventuate several disturbances that may lead to the development of neurodegenerative changes.
Therefore, compact, reliable approaches that allow the optical detection of Zn(2+) across the whole brain would contribute to our current understanding of the mechanisms that underlie neurological disease pathology. We developed an engineered fluorescence protein-based nanoprobe that can spatially and temporally resolve Zn(2+) in living zebrafish brain tissue.
The self-assembled engineered fluorescence protein on gold nanoparticles was shown to be confined to defined locations within the brain tissue, enabling site specific studies, compared to fluorescent protein-based molecular tools, which diffuse throughout the brain tissue. Two-photon excitation microscopy confirmed the physical and photometrical stability of these nanoprobes in living zebrafish (Danio rerio) brain tissue, while the addition of Zn(2+) quenched the nanoprobe fluorescence.
Combining orthogonal sensing methods with our engineered nanoprobes will enable the study of imbalances in homeostatic Zn(2+) regulation. The proposed bionanoprobe system offers a versatile platform to couple metal ion specific linkers and contribute to the understanding of neurological diseases.