Dataset Information

Using GFP video to track 3D movement and conditional gene expression in free-moving flies.

ABSTRACT: In vivo imaging and quantification of fluorescent reporter molecules is increasingly useful in biomedical research. For example, tracking animal movement in 3D with simultaneous quantification of fluorescent transgenic reporters allows for correlations between behavior, aging and gene expression. However implementation has been hindered in the past by the complexity of operating the systems.We report significant technical improvements and user-friendly software (called FluoreScore) that enables tracking of 3D movement and the dynamics of gene expression in adult Drosophila, using two cameras and recorded GFP videos. Expression of a transgenic construct encoding eGFP was induced in free-moving adult flies using the Gene-Switch system and RU486 drug feeding. The time course of induction of eGFP expression was readily quantified from internal tissues including central nervous tissue.FluoreScore should facilitate a variety of future studies involving quantification of movement behaviors and fluorescent molecules in free-moving animals.

SUBMITTER: Ardekani R

PROVIDER: S-EPMC3400653 | biostudies-other | 2012

REPOSITORIES: biostudies-other

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Using GFP video to track 3D movement and conditional gene expression in free-moving flies.

Ardekani Reza R Huang Yichuan Michelle YM Sancheti Prathamesh P Stanciauskas Ramunas R Tavaré Simon S Tower John J

PloS one 20120719 7

<h4>Background</h4>In vivo imaging and quantification of fluorescent reporter molecules is increasingly useful in biomedical research. For example, tracking animal movement in 3D with simultaneous quantification of fluorescent transgenic reporters allows for correlations between behavior, aging and gene expression. However implementation has been hindered in the past by the complexity of operating the systems.<h4>Results</h4>We report significant technical improvements and user-friendly software ...[more]

PMID: 22829875

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Project description:BackgroundAnimal movement expressed through home ranges or space-use can offer insights into spatial and habitat requirements. However, different classes of estimation methods are currently instinctively applied to answer home range, space-use or movement-based research questions regardless of their widely varying outputs, directly impacting conclusions. Recent technological advances in animal tracking (GPS and satellite tags), have enabled new methods to quantify animal space-use and movement pathways, but so far have primarily targeted mammal and avian species.MethodsMost reptile spatial ecology studies only make use of two older home range estimation methods: Minimum Convex Polygons (MCP) and Kernel Density Estimators (KDE), particularly with the Least Squares Cross Validation (LSCV) and reference (h ref ) bandwidth selection algorithms. These methods are frequently applied to answer space-use and movement-based questions. Reptile movement patterns are unique (e.g., low movement frequency, long stop-over periods), prompting investigation into whether newer movement-based methods -such as dynamic Brownian Bridge Movement Models (dBBMMs)- apply to Very High Frequency (VHF) radio-telemetry tracking data. We simulated movement data for three archetypical reptile species: a highly mobile active hunter, an ambush predator with long-distance moves and long-term sheltering periods, and an ambush predator with short-distance moves and short-term sheltering periods. We compared traditionally used estimators, MCP and KDE, with dBBMMs, across eight feasible VHF field sampling regimes for reptiles, varying from one data point every four daylight hours, to once per month.ResultsAlthough originally designed for GPS tracking studies, dBBMMs outperformed MCPs and KDE h ref across all tracking regimes in accurately revealing movement pathways, with only KDE LSCV performing comparably at some higher frequency sampling regimes. However, the LSCV algorithm failed to converge with these high-frequency regimes due to high site fidelity, and was unstable across sampling regimes, making its use problematic for species exhibiting long-term sheltering behaviours. We found that dBBMMs minimized the effect of individual variation, maintained low error rates balanced between omission (false negative) and commission (false positive), and performed comparatively well even under low frequency sampling regimes (e.g., once a month).ConclusionsWe recommend dBBMMs as a valuable alternative to MCP and KDE methods for reptile VHF telemetry data, for research questions associated with space-use and movement behaviours within the study period: they work under contemporary tracking protocols and provide more stable estimates. We demonstrate for the first time that dBBMMs can be applied confidently to low-resolution tracking data, while improving comparisons across regimes, individuals, and species.Supplementary informationSupplementary information accompanies this paper at 10.1186/s40462-020-00229-3.

Dataset Information

Using GFP video to track 3D movement and conditional gene expression in free-moving flies.

Publications

Using GFP video to track 3D movement and conditional gene expression in free-moving flies.

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