Ultrastructural comparison of dendritic spine morphology preserved with cryo and chemical fixation

  1. Hiromi Tamada
  2. Jerome Blanc
  3. Natalya Korogod
  4. Carl CH Petersen  Is a corresponding author
  5. Graham William Knott  Is a corresponding author
  1. Nagoya University, Japan
  2. Ecole Polytechnique Fédérale de Lausanne, Switzerland
  3. Haute École de Santé Vaud, Switzerland

Abstract

Previously we showed that cryo fixation of adult mouse brain tissue gave a truer representation of brain ultrastructure in comparison with a standard chemical fixation method (Korogod et al 2005). Extracellular space matched physiological measurements, there were larger numbers of docked vesicles and less glial coverage of synapses and blood capillaries. Here, using the same preservation approaches we compared the morphology of dendritic spines. We show that the length of the spine and the volume of its head is unchanged, however, the spine neck width is thinner by more than 30 % after cryo fixation. In addition, the weak correlation between spine neck width and head volume seen after chemical fixation was not present in cryo-fixed spines. Our data suggest that spine neck geometry is independent of the spine head volume, with cryo fixation showing enhanced spine head compartmentalization and a higher predicted electrical resistance between spine head and parent dendrite.

Data availability

All data generated during this study are included in the manuscript and the supporting files. Source data files are provided for all results. These are: Figures 1, 2, 3, 4 and 5 and Figure supplements for Figure 1 and 2.

Article and author information

Author details

  1. Hiromi Tamada

    Functional Anatomy and Neuroscience, Nagoya University, Nagoya, Japan
    Competing interests
    The authors declare that no competing interests exist.
  2. Jerome Blanc

    School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  3. Natalya Korogod

    School of health sciences, Haute École de Santé Vaud, Lausanne, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  4. Carl CH Petersen

    Laboratory of Sensory Processing, Brain Mind Institute, Faculty of Life Sciences, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
    For correspondence
    carl.petersen@epfl.ch
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3344-4495
  5. Graham William Knott

    School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
    For correspondence
    graham.knott@epfl.ch
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2956-9052

Funding

Swiss National Science Foundation (31003A_182010)

  • Carl CH Petersen

Swiss National Science Foundation (31003A_170082)

  • Graham William Knott

Japanese Society for the Promotion of Science (JP17K019)

  • Hiromi Tamada

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: This study was performed in strict accordance with the rules issued by the Swiss Federal Veterinary Office, under authorization 1889 issued by the 'Service de la consommation et des affaires vétérinaires' of the Canton de Vaud, Switzerland. The animals were handled according to approved institutional guidelines and under the experimentation license 1889.3 (Swiss Federal Veterinary Office).

Copyright

© 2020, Tamada et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 3,654
    views
  • 491
    downloads
  • 26
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Hiromi Tamada
  2. Jerome Blanc
  3. Natalya Korogod
  4. Carl CH Petersen
  5. Graham William Knott
(2020)
Ultrastructural comparison of dendritic spine morphology preserved with cryo and chemical fixation
eLife 9:e56384.
https://doi.org/10.7554/eLife.56384

Share this article

https://doi.org/10.7554/eLife.56384

Further reading

    1. Neuroscience
    Moritz F Wurm, Doruk Yiğit Erigüç
    Research Article

    Recognizing goal-directed actions is a computationally challenging task, requiring not only the visual analysis of body movements, but also analysis of how these movements causally impact, and thereby induce a change in, those objects targeted by an action. We tested the hypothesis that the analysis of body movements and the effects they induce relies on distinct neural representations in superior and anterior inferior parietal lobe (SPL and aIPL). In four fMRI sessions, participants observed videos of actions (e.g. breaking stick, squashing plastic bottle) along with corresponding point-light-display (PLD) stick figures, pantomimes, and abstract animations of agent–object interactions (e.g. dividing or compressing a circle). Cross-decoding between actions and animations revealed that aIPL encodes abstract representations of action effect structures independent of motion and object identity. By contrast, cross-decoding between actions and PLDs revealed that SPL is disproportionally tuned to body movements independent of visible interactions with objects. Lateral occipitotemporal cortex (LOTC) was sensitive to both action effects and body movements. These results demonstrate that parietal cortex and LOTC are tuned to physical action features, such as how body parts move in space relative to each other and how body parts interact with objects to induce a change (e.g. in position or shape/configuration). The high level of abstraction revealed by cross-decoding suggests a general neural code supporting mechanical reasoning about how entities interact with, and have effects on, each other.

    1. Neuroscience
    Magdalena Solyga, Georg B Keller
    Research Article

    Our movements result in predictable sensory feedback that is often multimodal. Based on deviations between predictions and actual sensory input, primary sensory areas of cortex have been shown to compute sensorimotor prediction errors. How prediction errors in one sensory modality influence the computation of prediction errors in another modality is still unclear. To investigate multimodal prediction errors in mouse auditory cortex, we used a virtual environment to experimentally couple running to both self-generated auditory and visual feedback. Using two-photon microscopy, we first characterized responses of layer 2/3 (L2/3) neurons to sounds, visual stimuli, and running onsets and found responses to all three stimuli. Probing responses evoked by audiomotor (AM) mismatches, we found that they closely resemble visuomotor (VM) mismatch responses in visual cortex (V1). Finally, testing for cross modal influence on AM mismatch responses by coupling both sound amplitude and visual flow speed to the speed of running, we found that AM mismatch responses were amplified when paired with concurrent VM mismatches. Our results demonstrate that multimodal and non-hierarchical interactions shape prediction error responses in cortical L2/3.