Biotechnology and Biological Sciences Research Council (BBSRC)
BB/M011151/1
英国
European Research Council (ERC)
695568 SYNNOVATE
European Union
Medical Research Council (MRC, United Kingdom)
MC_UP_1201/8
英国
Wellcome Trust
SBF005/1046
英国
引用
ジャーナル: Elife / 年: 2025 タイトル: The molecular infrastructure of glutamatergic synapses in the mammalian forebrain. 著者: Julia Peukes / Charlie Lovatt / Conny Leistner / Jerome Boulanger / Dustin R Morado / Martin J G Fuller / Wanda Kukulski / Fei Zhu / Noboru H Komiyama / John A G Briggs / Seth G N Grant / René A W Frank / 要旨: Glutamatergic synapses form the vast majority of connections within neuronal circuits, but how these subcellular structures are molecularly organized within the mammalian brain is poorly understood. ...Glutamatergic synapses form the vast majority of connections within neuronal circuits, but how these subcellular structures are molecularly organized within the mammalian brain is poorly understood. Conventional electron microscopy using chemically fixed, metal-stained tissue has identified a proteinaceous, membrane-associated thickening called the 'postsynaptic density' (PSD). Here, we combined mouse genetics and cryo-electron tomography to determine the 3D molecular architecture of fresh isolated and anatomically intact synapses in the adult forebrain. The native glutamatergic synapse did not consistently show a higher density of proteins at the postsynaptic membrane, thought to be characteristic of the PSD. Instead, a 'synaptoplasm' consisting of cytoskeletal elements, macromolecular complexes, and membrane-bound organelles extended throughout the pre- and post-synaptic compartments. Snapshots of active processes gave insights into membrane remodeling processes. Clusters of up to 60 ionotropic glutamate receptors were positioned inside and outside the synaptic cleft. Together, these information-rich tomographic maps present a detailed molecular framework for the coordinated activity of synapses in the adult mammalian brain.
凍結剤: ETHANE / チャンバー内温度: 277.15 K / 装置: FEI VITROBOT MARK IV
詳細
Mouse forebrain homogeneized in artificial cerebrospinal fluid (ACSF) to generate suspension of Psd95-GFP containing synaptoneurosomes. Samples was applied to cryoEM grid and plunge frozen in liquid ethane. CryET dataset collection was guided by Psd95-GFP cryoCLEM.
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電子顕微鏡法
顕微鏡
TFS KRIOS
特殊光学系
位相板: VOLTA PHASE PLATE / エネルギーフィルター - スリット幅: 20 eV
撮影
フィルム・検出器のモデル: GATAN K2 QUANTUM (4k x 4k) 平均電子線量: 1.0 e/Å2