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TitleA correlative workflow for synaptic imaging by cryo-electron tomography.
Journal, issue, pagesStructure, Year 2026
Publish dateMar 24, 2026
AuthorsThanh Thao Do / Anna Siegert / Florelle Domart / Fabienne Hahn / Christina Zeising / Sarah Muth / Constantin Pape / Kathrin Kusch / Thomas Dresbach / Silvio O Rizzoli / Arsen Petrovic / Rubén Fernández-Busnadiego /
PubMed AbstractDespite decades of intense research, the molecular organization of the synapse is not well understood. To address this issue, we sought to develop a method for systematic imaging of synapses by cryo- ...Despite decades of intense research, the molecular organization of the synapse is not well understood. To address this issue, we sought to develop a method for systematic imaging of synapses by cryo-electron tomography (cryo-ET), a technology capable of mapping cellular architecture at molecular resolution. Thinning of cellular samples by cryo-focused ion beam milling is a prerequisite for high-quality cryo-ET imaging, but this process needs to be guided to the structures of interest. To allow synaptic targeting, we established a correlative cryo-light/electron microscopy approach by which synapses are fluorescently labeled in a minimally invasive manner, using a synthetic binder of the postsynaptic scaffold PSD-95 and antibodies against the presynaptic protein synaptotagmin-1. Cryo-ET imaging at sites of colocalization predominantly revealed excitatory synapses. Our method allows structural studies of synapse-resident protein complexes in situ, facilitating investigations of the molecular architecture of synapses.
External linksStructure / PubMed:41881019
MethodsEM (tomography) / EM (subtomogram averaging)
Resolution10.0 Å
Structure data

EMDB-54384: In situ cryo-electron tomogram of rat hippocampal synapse
Method: EM (tomography)

EMDB-54398: In situ cryo-electron tomogram of rat hippocampal synapse
Method: EM (tomography)

EMDB-54405: In situ cryo-electron tomogram of rat hippocampal synapse
Method: EM (tomography)

EMDB-54406: In situ cryo-electron tomogram of rat hippocampal synapse
Method: EM (tomography)

EMDB-54407: In situ cryo-electron tomogram of rat hippocampal synapse
Method: EM (tomography)

EMDB-54420: In situ cryo-electron tomogram of rat hippocampal synapse
Method: EM (tomography)

EMDB-54421: In situ cryo-electron tomogram of rat hippocampal synapse
Method: EM (tomography)

EMDB-54422: In situ cryo-electron tomogram of rat hippocampal synapse
Method: EM (tomography)

EMDB-54424: In situ cryo-electron tomogram of rat hippocampal synapse
Method: EM (tomography)

EMDB-54432: In situ cryo-electron tomogram of rat hippocampal synapse
Method: EM (tomography)

EMDB-54447: Subtomogram average of synaptic microtubule from rat hippocampal neurons
Method: EM (subtomogram averaging) / Resolution: 10.0 Å

Source
  • Rattus norvegicus (Norway rat)

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