Resolving the molecular architecture of the photoreceptor active zone with 3D-MINFLUX.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 35857490.
- Also identified by DOI 10.1126/sciadv.abl7560 and PMC identifier 9286502.
- Licence recorded as CC BY-NC.
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Abstract
Cells assemble macromolecular complexes into scaffoldings that serve as substrates for catalytic processes. Years of molecular neurobiology research indicate that neurotransmission depends on such optimization strategies. However, the molecular topography of the presynaptic active zone (AZ), where transmitter is released upon synaptic vesicle (SV) fusion, remains to be visualized. Therefore, we implemented MINFLUX optical nanoscopy to resolve the AZ of rod photoreceptors. This was facilitated by a novel sample immobilization technique that we name heat-assisted rapid dehydration (HARD), wherein a thin layer of rod synaptic terminals (spherules) was transferred onto glass coverslips from fresh retinal slices. Rod ribbon AZs were readily immunolabeled and imaged in 3D with a precision of a few nanometers. Our 3D-MINFLUX results indicate that the SV release site in rods is a molecular complex of bassoon-RIM2-ubMunc13-2-Ca<sub>v</sub>1.4, which repeats longitudinally on both sides of the ribbon.