Source 1primary paper2017Proceedings of the National Academy of Sciences
Toolkit/Tric-LUC
Tric-LUC
Also known as: TRIC-LUC reporter
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
To this end, we adapted a GFP-based calcium reporter to luciferase that was expressed in small subsets of circadian neurons. This Tric-LUC reporter allowed a direct comparison of luciferase activity with locomotor activity, which was assayed in the same flies with video recording.
Usefulness & Problems
Why this is useful
Tric-LUC is a luciferase-adapted calcium reporter used to profile activity of specific circadian neurons in freely moving flies. It produces luciferase signals that can be compared against behavior in the same animals.; temporal calcium profiling of specific circadian neurons in freely moving flies; comparing neuronal reporter output with locomotor activity in the same flies
Source:
Tric-LUC is a luciferase-adapted calcium reporter used to profile activity of specific circadian neurons in freely moving flies. It produces luciferase signals that can be compared against behavior in the same animals.
Source:
temporal calcium profiling of specific circadian neurons in freely moving flies
Source:
comparing neuronal reporter output with locomotor activity in the same flies
Problem solved
It addresses the lack of general methods for assessing calcium profiles of specific neurons in freely moving flies.; lack of general methods for reliably assessing calcium profiles of specific neurons in freely moving flies
Source:
It addresses the lack of general methods for assessing calcium profiles of specific neurons in freely moving flies.
Source:
lack of general methods for reliably assessing calcium profiles of specific neurons in freely moving flies
Problem links
lack of general methods for reliably assessing calcium profiles of specific neurons in freely moving flies
LiteratureIt addresses the lack of general methods for assessing calcium profiles of specific neurons in freely moving flies.
Source:
It addresses the lack of general methods for assessing calcium profiles of specific neurons in freely moving flies.
Published Workflows
Objective: Develop a method to assess calcium profiles of specific circadian neurons in freely moving flies and compare those profiles with behavior in the same animals.
Why it works: The workflow is expected to work because luciferase output from the adapted reporter can be recorded in freely moving flies and directly compared with locomotor behavior measured in the same individuals.
neuronal calcium reportingpossible reflection of firing profilesreporter adaptation from GFP-based calcium reporter to luciferasesimultaneous luciferase and video-based locomotor recording
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into 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
The reporter must be expressed in selected circadian neuron subsets, and experiments require luciferase recording together with video recording of locomotor activity in the same flies.; requires expression in specific neuron subsets; requires luciferase recording and video-based locomotor activity measurement in the same flies
From the abstract alone, it does not provide a guaranteed direct measurement of firing, because the authors only suggest that luciferase profiles reflect calcium and in some cases firing profiles.; the abstract frames luciferase profiles as reflecting neuronal calcium and in some cases firing profiles, indicating indirect readout rather than direct electrophysiology
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Tric-LUC allowed direct comparison of luciferase activity with locomotor activity in the same flies using video recording.
In E cells, the Tric-LUC luciferase profile paralleled evening locomotor activity.
In glutamatergic DN1s, the Tric-LUC luciferase profile paralleled daytime sleep.
The authors suggest that luciferase profiles from E cells and gDN1s reflect neuronal calcium and in some cases firing profiles in wake-behaving flies.
Tric-LUC enabled temporal calcium profiling of specific circadian neurons in freely moving flies by adapting a GFP-based calcium reporter to luciferase.
Approval Evidence
1 source5 linked approval claimsfirst-pass slug tric-luc
To this end, we adapted a GFP-based calcium reporter to luciferase that was expressed in small subsets of circadian neurons. This Tric-LUC reporter allowed a direct comparison of luciferase activity with locomotor activity, which was assayed in the same flies with video recording.
Source:
assay capabilitysupports
Tric-LUC allowed direct comparison of luciferase activity with locomotor activity in the same flies using video recording.
Source:
behavioral correlationsupports
In E cells, the Tric-LUC luciferase profile paralleled evening locomotor activity.
Source:
behavioral correlationsupports
In glutamatergic DN1s, the Tric-LUC luciferase profile paralleled daytime sleep.
Source:
interpretive mechanismsupports
The authors suggest that luciferase profiles from E cells and gDN1s reflect neuronal calcium and in some cases firing profiles in wake-behaving flies.
Source:
tool capabilitysupports
Tric-LUC enabled temporal calcium profiling of specific circadian neurons in freely moving flies by adapting a GFP-based calcium reporter to luciferase.
Source:
Comparisons
Source-stated alternatives
The abstract mentions similar profiles from novel reporters based on transcription factor activation, but does not name them in the source text provided.
Source:
The abstract mentions similar profiles from novel reporters based on transcription factor activation, but does not name them in the source text provided.
Source-backed strengths
enables direct comparison of luciferase activity with locomotor activity in the same animals; can be expressed in small subsets of circadian neurons
Source:
enables direct comparison of luciferase activity with locomotor activity in the same animals
Source:
can be expressed in small subsets of circadian neurons
Compared with CheRiff
Tric-LUC and CheRiff address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
Compared with intermolecular disulfide-based light switch
Tric-LUC and intermolecular disulfide-based light switch address a similar problem space.
Shared frame: same top-level item type
Strengths here: looks easier to implement in practice.
Tric-LUC and Pyr-NHS-functionalised 3D graphene foam electrode biosensor address a similar problem space.
Shared frame: same top-level item type
Ranked Citations
- 1.