Single-particle tracking of NMDA receptors at hippocampal synapses

Lecturer: Mgr. Petra Zahumenská / Department of Neurochemistry

Annotation: N-methyl-D-aspartate receptors (NMDARs) are ionotropic glutamate receptors essential for excitatory neurotransmission. These heterotetrameric complexes consist of two obligatory GluN1 subunits combined with two GluN2 (GluN2A-GluN2D) or GluN3 (GluN3A-GluN3B) subunits. The surface expression and subtype composition of NMDARs are tightly regulated through externalization, internalization, and lateral diffusion between synaptic and extrasynaptic regions.

In our study, we utilized the uPAINT (Universal Point Accumulation Imaging in Nanoscale Topography) microscopy technique to examine the surface dynamics of different NMDAR subtypes. uPAINT enables the tracking of individual NMDARs with a high spatial and temporal resolution by labelling them with fluorescent probes. This allows us to observe the distinct patterns of synaptic and extrasynaptic localization of GluN2A- and GluN2B-containing NMDARs in hippocampal neurons. Our approach combines the use of a small anti-GFP nanobody derived from a llama conjugated with an Atto647N fluorophore, along with the tdTomato-Homer1c marker, to define synaptic regions. This allows us to gain insights into the mobility and distribution of NMDAR subtypes at the synaptic level.

The seminar will take place on 3. 10. 2024 from 2 p.m. in the Turquoise Auditorium of the IEM CAS.

We look forward to your participation.