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Home > Industry Information > Quantum dot tracing technique reveals the dynamic properties of nerve growth factor receptor

Quantum dot tracing technique reveals the dynamic properties of nerve growth factor receptor

2021-05-17

During the regulation of the nervous system, specific ligands bind to neuronal membrane surface receptors, which in turn activates intracellular and downstream signaling. This process is important for maintaining normal nervous system function, with abnormalities in many neurological dysfunctional diseases such as Alzheimer's disease and mental illnesses such as depression. The Tania Q. Vu group of Oregon Health and Technology University in the United States used Quantum dots (QDs) tracer technology to observe the brain-derived neurotrophic factor (BDNF) and TrkB receptors in neurons in real time. The transport process of body binding and endocytosis into the cytoplasm.

Anke Vermehren-Schmaedick et al. biotinylated BDNF and incubated with streptavidin-coupled quantum dots to obtain the quantum dot marker QD-BDNF. The in vitro ELISA assay confirmed that the coupling of BDNF to quantum dots did not affect its binding and activation to the TrkB receptor. In the observation of neuronal cell in vivo imaging, QD-BDNF specifically binds to TrkB and induces TrkB-mediated signaling. Moreover, QD-BDNF complex can undergo long-term imaging observation, and the imaging effect is better than Alexa-BDNF. And there was no adverse effect on cell morphology under continuous excitation conditions (Figure 1). The above researchers used quantum dot single-molecule imaging technology to directly observe the BDNF-TrkB complex in vivo into the cytoplasm and return to the cell membrane recycling process (Figure 2), thus confirming the speculation of the relevant kinetics.

Figure 1 Quantum dot-labeled brain-derived neurotrophic factor (QD-BDNF) binds to the TrkB receptor on neurons and shows good photophysical properties.

(A) Compared to traditional fluorescent dye markers (Alexa-BDNF, 25 nM), quantum dot markers (QD-BDNF, 200 pM) showed higher sensitivity;

(B) Scintillation analysis of two QD-BDNFs in one neuronal cell;

(C) For high-resolution spatial detection of QD-BDNF, accurate display of cell membrane or cytoplasmic localization;

(D) Analysis of fluorescence intensity showed that the quantum dots showed good fluorescence stability and did not adversely affect cell morphology under continuous excitation conditions.

Figure 2 QD-BDNF is endocytosed and recovered in neuronal cell bodies.

Left: QD-BDNF is endocytosed to the cytoplasm after binding to the receptor on the cell membrane; right: QD-BDNF moves from the cytoplasm to the cell membrane and returns to the cell membrane.

Source of the document:

Vermehren-Schmaedick A, Krueger W, Jacob T, Ramunno-Johnson D, Balkowiec A, Lidke KA, Vu TQ. Heterogeneous intracellular trafficking dynamics of brain-derived neurotrophic factor complexes in the neuronal soma revealed by single quantum dot tracking. PLoS One. 2014;9(4):e95113.


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