Source 1primary paper2013Proceedings of the National Academy of Sciences
Toolkit/genetically encoded indicators
genetically encoded indicators
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Genetically encoded indicators and antineoepitope antibodies should improve spatial and temporal resolution of the in vivo activity of proteases that locally erode PNN.
Usefulness & Problems
Why this is useful
Genetically encoded indicators are proposed to improve spatial and temporal resolution when monitoring in vivo activity of proteases that locally erode PNN.; improving spatial and temporal resolution of in vivo protease activity
Source:
Genetically encoded indicators are proposed to improve spatial and temporal resolution when monitoring in vivo activity of proteases that locally erode PNN.
Source:
improving spatial and temporal resolution of in vivo protease activity
Problem solved
They are suggested as a way to better resolve when and where PNN-eroding proteases are active.; proposed monitoring of proteases that locally erode PNN
Source:
They are suggested as a way to better resolve when and where PNN-eroding proteases are active.
Source:
proposed monitoring of proteases that locally erode PNN
Problem links
proposed monitoring of proteases that locally erode PNN
LiteratureThey are suggested as a way to better resolve when and where PNN-eroding proteases are active.
Source:
They are suggested as a way to better resolve when and where PNN-eroding proteases are active.
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.
Mechanisms
protease activity reportingTechniques
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 genetically-encoded-indicators
Genetically encoded indicators and antineoepitope antibodies should improve spatial and temporal resolution of the in vivo activity of proteases that locally erode PNN.
Source:
proposed method capabilitysupports
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.
Source:
Comparisons
Source-stated alternatives
The abstract pairs these indicators with antineoepitope antibodies as another route to improved resolution.
Source:
The abstract pairs these indicators with antineoepitope antibodies as another route to improved resolution.
Source-backed strengths
should improve spatial and temporal resolution in vivo
Source:
should improve spatial and temporal resolution in vivo
Compared with antineoepitope antibodies
The abstract pairs these indicators with antineoepitope antibodies as another route to improved resolution.
Shared frame: source-stated alternative in extracted literature
Strengths here: should improve spatial and temporal resolution in vivo.
Source:
The abstract pairs these indicators with antineoepitope antibodies as another route to improved resolution.
Ranked Citations
- 1.