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#structbio

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Stephen Royle

Looks like cryo-ET studies of synapses are hot right now (pardon the pun). Another preprint, this time on dopaminergic terminals, looking at vesicle structures in situ.

biorxiv.org/content/10.1101/20

bioRxiv · Ultrastructural Dynamics of Dopaminergic Presynaptic Release Sites revealed by Cryo-correlative Light and Electron MicroscopyDopaminergic neurons are fundamental in governing motivation, movement, and many aspects of cognition. The targeted modulation of dopaminergic signaling serves as a cornerstone in developing therapeutic interventions for conditions such as Parkinson's disease, schizophrenia, and addiction. Despite the pivotal role of dopaminergic neurons, the ultrastructure and associated dynamics of dopaminergic synapses remain poorly understood. Here, we develop and utilize a cryo-correlative light and electron microscopy process chain to investigate the micro- to nanoscale architecture and organelle content of dopaminergic presynaptic release sites. Using cryo electron tomography, we identify several protein complexes crucial to dopaminergic function and we utilize subtomogram averaging to resolve in situ assemblies of the TRiC/CCT chaperone and vacuolar-type ATPase. Lastly, we find that pharmacological treatments using either dopamine or the dopamine D2 receptor antagonist, haloperidol, bidirectionally modulate vesicular content, mitochondrial size and calcium phosphate deposition. These findings contribute to our general understanding of the composition and ultrastructural dynamics of dopaminergic presynaptic release sites and provide a methodological platform for further studies of the structure and cell biology of dopaminergic neurons and their responses. ### Competing Interest Statement The authors have declared no competing interest.
#CellBiology#structbio
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Public
Stephen Royle

and while we are at it, this other preprint uses cryo-ET on *native* synapses to look at vesicles in the terminal

biorxiv.org/content/10.1101/20
#structuralbiology #structbio #cellbiology

bioRxiv · Cryo-electron tomographic investigation of native hippocampal glutamatergic synapsesChemical synapses are the major sites of communication between neurons in the nervous system and mediate either excitatory or inhibitory signaling [1]. At excitatory synapses, glutamate is the primary neurotransmitter and upon release from presynaptic vesicles, is detected by postsynaptic glutamate receptors, which include ionotropic AMPA and NMDA receptors [2, 3]. Here we have developed methods to identify glutamatergic synapses in brain tissue slices, label AMPA receptors with small gold nanoparticles (AuNPs), and prepare lamella for cryo-electron tomography studies. The targeted imaging of glutamatergic synapses in the lamella is facilitated by fluorescent pre- and postsynaptic signatures, and the subsequent tomograms allow for identification of key features of chemical synapses, including synaptic vesicles, the synaptic cleft and AuNP-labeled AMPA receptors. These methods pave the way for imaging natively derived brain regions at high resolution, using unstained, unfixed samples preserved under near-native conditions. ### Competing Interest Statement The authors have declared no competing interest.
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