Artikel
High-pressure freezing for EM to study plasticity and pathology of identified central synapses
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Autoren
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Michael Frotscher
- Center for Molecular Neurobiology Hamburg, Structural Neurobiology, Hamburg, Germany
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Daniel Studer
- University of Bern, Institute of Anatomy, Bern, Switzerland
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Shanting Zhao
- Center for Molecular Neurobiology Hamburg, Structural Neurobiology, Hamburg, Germany
| Veröffentlicht: | 25. August 2015 |
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Gliederung
Text
Question: Electron microscopy is the best way to visualize synapses with their presynaptic and postsynaptic compartments including structural specifications such as synaptic vesicles, active zones, and the spine apparatus. It is an open question to what extent the structure and distribution of these compartments are preserved when the tissue is subjected to conventional aldehyde fixation and dehydration procedures.
Methods: Here, we used high-pressure freezing (HPF) and cryosubstitution to immobilize hippocampal tissue within 50 ms without the use of aldehyde solutions and dehydration in ascending series of ethanol.
Results: HPF resulted in an excellent preservation of synaptic ultrastructure. Induction of long-term potentiation (LTP) prior to shock freezing induced the formation of new spines and synaptic contacts at identified mossy fiber synapses in the hippocampus accompanied by quantitative changes in the immunogold labeling for p-cofilin.
Conclusions: HPF allows for the the study of subtle structural synaptic changes associated with functional synaptic plasticity.
Figure 1 [Fig. 1]
