Source 1primary paper2013Proceedings of the National Academy of Sciences
Toolkit/serial-section EM
serial-section EM
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The 3D intertwining of PNN and synapses would be imaged by serial-section EM.
Usefulness & Problems
Why this is useful
Serial-section EM is proposed to image the 3D intertwining of perineuronal nets and synapses.; imaging 3D intertwining of perineuronal net and synapses
Source:
Serial-section EM is proposed to image the 3D intertwining of perineuronal nets and synapses.
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imaging 3D intertwining of perineuronal net and synapses
Problem solved
It is suggested as a way to directly visualize the structural arrangement underlying the hole-pattern memory hypothesis.; proposed visualization of PNN hole architecture relative to synapses
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It is suggested as a way to directly visualize the structural arrangement underlying the hole-pattern memory hypothesis.
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proposed visualization of PNN hole architecture relative to synapses
Problem links
proposed visualization of PNN hole architecture relative to synapses
LiteratureIt is suggested as a way to directly visualize the structural arrangement underlying the hole-pattern memory hypothesis.
Source:
It is suggested as a way to directly visualize the structural arrangement underlying the hole-pattern memory hypothesis.
Published Workflows
Objective: Test the hypothesis that very long-term memories are stored as the pattern of holes in the perineuronal net by combining structural imaging, component lifetime measurements, in vivo monitoring of PNN-eroding proteases, perturbation approaches, and activity-linked functional manipulation.
Why it works: The proposed workflow combines complementary measurements: structural imaging to visualize PNN-synapse geometry, isotope-based approaches to compare component lifetimes, in vivo reporters to resolve protease activity, and perturbation plus behavioral comparison to test functional importance.
very long-term memories stored in the pattern of holes in the perineuronal netprotease-mediated local erosion of PNNrelationship between PNN structure and synapse formationserial-section EMpulse-chase 15N labeling14C content analysis in human cadaver brainsgenetically encoded indicatorsantineoepitope antibodiesgenetic knockoutspharmacological inhibitorsgenetically encoded snapshot reporterimaging of biosynthesis of PNN components and proteases
Stages
- 1.Structural imaging of PNN-synapse architecture(functional_characterization)
To directly visualize the structural arrangement proposed to encode very long-term memory.
Selection: Image the 3D intertwining of PNN and synapses
- 2.Component lifetime comparison(secondary_characterization)
To test whether PNN components have lifetimes consistent with a substrate for very long-term memory.
Selection: Compare lifetimes of PNN versus intrasynaptic components
- 3.In vivo protease activity resolution(functional_characterization)
To better resolve when and where proteases locally erode PNN in vivo.
Selection: Improve spatial and temporal resolution of in vivo activity of proteases that locally erode PNN
- 4.Perturbation and functional testing of PNN erosion(confirmatory_validation)
To causally test whether manipulating PNN erosion and the implicated neuronal ensembles changes behavior.
Selection: Use genetic, pharmacological, and activity-linked perturbation tools to test functional importance and behavioral consequences
- 5.Imaging biosynthesis of PNN components and proteases(secondary_characterization)
To observe production of PNN components and proteases relevant to net maintenance and remodeling.
Selection: Image biosynthesis of PNN components and proteases
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Techniques
Functional AssayTarget processes
No target processes tagged yet.
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
Operational role: sensor. Implementation mode: genetically encoded. Cofactor status: cofactor requirement unknown.
Independent follow-up evidence is still limited. Validation breadth across biological contexts is still narrow. Independent reuse still looks limited, so the evidence base may be fragile. No canonical validation observations are stored yet, so context-specific performance remains under-specified.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
14C content in human cadaver brains is proposed to compare lifetimes of PNN and intrasynaptic components.
and (14)C content in human cadaver brains
Genetically encoded indicators and antineoepitope antibodies should improve spatial and temporal resolution of in vivo activity of proteases that locally erode PNN.
Genetically encoded indicators and antineoepitope antibodies should improve spatial and temporal resolution of the in vivo activity of proteases that locally erode PNN.
Pulse-chase 15N labeling in mice is proposed to compare lifetimes of PNN and intrasynaptic components.
Lifetimes of PNN vs. intrasynaptic components would be compared with pulse-chase (15)N labeling in mice
Serial-section EM is proposed to image the 3D intertwining of perineuronal net and synapses.
The 3D intertwining of PNN and synapses would be imaged by serial-section EM.
A genetically encoded snapshot reporter is proposed to capture neuronal activity patterns at an illumination-defined time, mark those cells by driving effector gene expression, and enable selective excitation, inhibition, or ablation to test functional importance.
a genetically encoded snapshot reporter, which will capture the pattern of activity throughout a large ensemble of neurons at a time precisely defined by the triggering illumination, drive expression of effector genes to mark those cells, and allow selective excitation, inhibition, or ablation to test their functional importance
The genetically encoded snapshot reporter should enable more precise inhibition or potentiation of PNN erosion to compare with behavioral consequences.
The snapshot reporter should enable more precise inhibition or potentiation of PNN erosion to compare with behavioral consequences.
Approval Evidence
1 source1 linked approval claimfirst-pass slug serial-section-em
The 3D intertwining of PNN and synapses would be imaged by serial-section EM.
Source:
proposed method capabilitysupports
Serial-section EM is proposed to image the 3D intertwining of perineuronal net and synapses.
The 3D intertwining of PNN and synapses would be imaged by serial-section EM.
Source:
Comparisons
Source-backed strengths
provides 3D structural imaging of PNN-synapse relationships
Source:
provides 3D structural imaging of PNN-synapse relationships
Compared with Langendorff perfused heart electrical recordings
serial-section EM and Langendorff perfused heart electrical recordings address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
Compared with native green gel system
serial-section EM and native green gel system address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
serial-section EM and sub-picosecond pump-probe analysis of bacteriorhodopsin pigments address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
Ranked Citations
- 1.