Computer assisted detection of axonal bouton structural plasticity in in vivo time-lapse images

Abstract
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Abstract

The ability to measure minute structural changes in neural circuits is essential for long-term in vivo imaging studies. Here, we propose a methodology for detection and measurement of structural changes in axonal boutons imaged with time-lapse two-photon laser scanning microscopy (2PLSM). Correlative 2PLSM and 3D electron microscopy (EM) analysis, performed in mouse barrel cortex, showed that the proposed method has low fractions of false positive/negative bouton detections (2/0 out of 18), and that 2PLSM-based bouton weights are correlated with their volumes measured in EM (r=0.93). Next, the method was applied to a set of axons imaged in quick succession to characterize measurement uncertainty. The results were used to construct a statistical model in which bouton addition, elimination, and size changes are described probabilistically, rather than being treated as deterministic events. Finally, we demonstrate that the model can be used to quantify significant structural changes in boutons in long-term imaging experiments.

Data availability

The following data sets were generated

1. Gala R
2. Lebrecht D
3. Sahlender DA
4. Jorstad A
5. Knott G
6. Holtmaat A
7. Stepanyants A
(2017) Data from: Computer assisted detection of axonal bouton structural plasticity in in vivo time-lapse images
Available at Dryad Digital Repository under a CC0 Public Domain Dedication.

http://dx.doi.org/10.5061/dryad.3q50t

Article and author information

Author details

Rohan Gala

Department of Physics, Northeastern University, Boston, United States

Competing interests
The authors declare that no competing interests exist.
Daniel Lebrecht

Department of Basic Neurosciences, Faculty of Medicine, University of Geneva, Geneva, Switzerland

Competing interests
The authors declare that no competing interests exist.
Daniela A Sahlender

Biological Electron Microscopy Facility, Centre of Electron Microscopy, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland

Competing interests
The authors declare that no competing interests exist.
Anne Jorstad

Biological Electron Microscopy Facility, Centre of Electron Microscopy, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-6438-1979
Graham Knott

Biological Electron Microscopy Facility, Centre of Electron Microscopy, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-2956-9052
Anthony Holtmaat

Department of Basic Neurosciences, Faculty of Medicine, University of Geneva, Geneva, Switzerland

Competing interests
The authors declare that no competing interests exist.
Armen Stepanyants

Department of Physics, Northeastern University, Boston, United States

For correspondence
a.stepanyants@neu.edu

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-9387-2320

Funding

National Institutes of Health (R01 NS091421)

Armen Stepanyants

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (331003A_153448)

Anthony Holtmaat

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (CRSII3-154453)

Graham Knott
Anthony Holtmaat

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (51NF40_158776)

Anthony Holtmaat

International Foundation for Research in Paraplegia (Chair Alain Rossier)

Anthony Holtmaat

Air Force Office of Scientific Research (FA9550-15-1-0398)

Armen Stepanyants

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

Ethics

Animal experimentation: All experiments were performed according to the guidelines of the Swiss Federal Act on Animal Protection and Swiss Animal Protection Ordinance. The ethics committee of the University of Geneva and the Cantonal Veterinary Office of Geneva, Switzerland (approval code GE/61/17) approved all experiments.

Copyright

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.