Super-resolution 2-photon microscopy reveals that the morphology of each dendritic spine correlates with diffusive but not synaptic properties
Open accessAbstract
The structure of dendritic spines suggests a specialized function in compartmentalizing synaptic signals near active synapses. Indeed, theoretical and experimental analyses indicate that the diffusive resistance of the spine neck is sufficient to effectively compartmentalize some signaling molecules in a spine for the duration of their activated lifetime. Here we describe the application of 2-photon microscopy combined with stimulated emission depletion (STED-2P) to the biophysical study of the relationship between synaptic signals and spine morphology, demonstrating the utility of combining STED-2P with modern optical and electrophysiological techniques. Morphological determinants of fluorescence recovery time were identified and evaluated within the context of a simple compartmental model describing diffusive transfer between spine and dendrite. Correlations between the neck geometry and the amplitude of synaptic potentials and calcium transients evoked by 2-photon glutamate uncaging were also investigated.
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Recent breakthroughs in fluorescence microscopy allow imaging at resolutions below the diffraction limit, allowing to directly explore how spine shape affects diffusion of cytoplasmic or membrane-embedded molecules.
“Recent breakthroughs in fluorescence microscopy allow imaging at resolutions below the diffraction limit, allowing to directly explore how spine shape affects diffusion of cytoplasmic or membrane-embedded molecules (Takasaki and Sabatini, 2014; Tønnesen et al., 2014).”
From Barriers in the brain: resolving dendritic spine morphology and compartmentalization (2014) · cites this paper as Takasaki and Sabatini, 2014
The Introduction reports that STED-2P achieves ~50 nm resolution, and the Results describe combining spine imaging with fluorescence recovery measurements to examine diffusion of a cytoplasmic dye. The paper does not report examining diffusion of membrane-embedded molecules, so that part of the sentence is not supported by this source. (LLM, conf=0.86)
Experimental — accuracy not yet measuredTakasaki and Sabatini find a weak decrease of recovery time with spine head volume, whereas the equation suggests a linear increase.
“However, whereas the equation suggests a linear increase of the recovery time scale with spine head volume, Takasaki and Sabatini (2014) instead find a weak decrease.”
From Barriers in the brain: resolving dendritic spine morphology and compartmentalization (2014) · cites this paper as Takasaki and Sabatini, 2014
The Modeling of diffusive compartmentalization section gives equation (3), in which recovery time increases linearly with spine head volume when the other parameters are held fixed. In Results (Figure 2E), the measured association is weakly negative (r = −0.19), but it is not statistically significant (p = 0.26). Describing it as a decrease the authors ‘find’ overstates that result. (LLM, conf=0.86)
Experimental — accuracy not yet measuredElectric compartmentalization is moderate, but not absent, in most spines.
“Most recently, STED microscopy on dendritic spines in organotypic and acute slices suggested that electric compartmentalization is moderate, but not absent, in most spines (Takasaki and Sabatini, 2014; Tønnesen et al., 2014).”
From Barriers in the brain: resolving dendritic spine morphology and compartmentalization (2014) · cites this paper as Takasaki and Sabatini, 2014
The Introduction reports evidence of significant voltage drops across spine necks, supporting the suggestion that electrical compartmentalization is not absent. But the Discussion (‘Correlation of morphology and synaptic signals’; page unavailable) says this study found no significant correlation between spine dimensions and synaptic potentials, and it does not establish that compartmentalization is moderate in most spines. Its STED-2P analysis was limited to acute brain slices. (LLM, conf=0.84)
Experimental — accuracy not yet measuredA fraction of spines strongly deviated from the average trends, suggesting that small local constrictions, local protein accumulations and organelle positioning in the neck may create additional diffusion barriers.
“Interestingly, a fraction of spines strongly deviated from the average trends that were observed (Takasaki and Sabatini, 2014), suggesting that small local constrictions, local protein accumulations and organelle positioning in the neck may create additional diffusion barriers (Arellano et al., 2007; Yuste, 2013).”
From Barriers in the brain: resolving dendritic spine morphology and compartmentalization (2014) · cites this paper as Takasaki and Sabatini, 2014
There is an inverse relation between spine neck diameter d and recovery time, which appears to follow the predicted inverse quadratic relation (τ ∝ 1/d2).
“Indeed, both groups find an inverse relation between spine neck diameter d and the recovery time, which appears to follow the predicted inverse quadratic relation (τ ∝ 1/d2).”
From Barriers in the brain: resolving dendritic spine morphology and compartmentalization (2014) · cites this paper as Takasaki and Sabatini, 2014
The recovery time scale τ roughly follows what would be expected if diffusion is governed largely by spine geometry: τ=V*L/(D*A).
“Both studies suggest that the recovery time scale τ roughly follows what would be expected if diffusion is governed largely by spine geometry: τ=V*LD*A”
From Barriers in the brain: resolving dendritic spine morphology and compartmentalization (2014) · cites this paper as Takasaki and Sabatini, 2014
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- Authors
- Takasaki, K., & Sabatini, B. L.
- Affiliations
- Howard Hughes Medical Institute (US)
- Corresponding author
- Sabatini, Bernardo L.
- Venue
- Frontiers in Neuroanatomy
- Publisher
- Frontiers Media SA
- Published
- 2014-05-07
- Volume
- 8
- Pages
- 29-29
- ISSN
- 1662-5129 (ISSN-L)
- PMC
- PMC4019874
- OpenAlex
- W2129084669
- Cited by
- 51
- References
- 34
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 1 · 2023: 5 · 2022: 3 · 2021: 5 · 2020: 2 · 2019: 7 · 2018: 3 · 2017: 6 · 2016: 5 · 2015: 7 · 2014: 3
- FWCI
- 3.20
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Frontiers in Neuroanatomy (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- National Institute of Neurological Disorders and Stroke (RC1NS068917)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Neuroscience › Cognitive Neuroscience › Neural dynamics and brain function
- Keywords
- dendritic spines, dendritic spine morphology, glutamate uncaging, calcium transients, synaptic potentials
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