Measuring Glucose Uptake in Drosophila Models of TDP-43 Proteinopathy

作者： Suvithanandhini Loganathan*1, Hannah E. Ball*1, Ernesto Manzo1,2, Daniela C. Zarnescu1,3 1Department of Molecular and Cellular Biology,University of Arizona, Tucson, 2Vollum Institute,Oregon Health and Science University, 3Department of Neuroscience,University of Arizona, Tucson

简介：

Overview

This study investigates glucose uptake in Drosophila motor neurons affected by TAR DNA binding protein (TDP-43) proteinopathy using a FRET-based, genetically encoded glucose sensor. The technique allows for real-time measurement of intracellular glucose levels, highlighting metabolic changes in neurons related to neurodegenerative conditions.

Key Study Components

Area of Science

• Neuroscience
• Cell Biology
• Neurodegeneration

Background

• TDP-43 is associated with neurodegenerative diseases such as ALS.
• Understanding glucose metabolism is critical for insights into neuronal health.
• This study utilizes a FRET-based sensor for dynamic measurements in living tissues.
• Previous methods may not have allowed for real-time analysis within specific neuron types.

Purpose of Study

• To measure changes in glucose uptake in motor neurons under TDP-43 proteinopathy.
• To compare glucose metabolism between ALS models and control neurons.
• To utilize a live imaging approach for direct observations of metabolic changes.

Methods Used

• Live imaging of Drosophila motor neurons using a FRET-based glucose sensor.
• Dissections were performed on wandering third instar larvae to prepare neuronal samples.
• Image acquisition utilized a confocal microscope to capture changes every 10 seconds.
• FRET efficiency was calculated to determine glucose uptake in response to stimulation.

Main Results

• Control motor neurons exhibited a small increase in glucose uptake, while TDP-43 mutant neurons showed a significantly higher increase.
• FRET by CFP ratios indicated alterations in intracellular glucose concentrations upon stimulation.
• No baseline differences were observed between genotypes, emphasizing the impact of glucose stimulation.
• Results suggest critical differences in metabolic responses between healthy and impaired neurons.

Conclusions

• This study demonstrates the utility of FRET-based sensors in assessing metabolic changes in live neurons.
• The findings provide insights into glucose metabolism's role in neuronal health and disease.
• Implications for understanding metabolic dysregulation in neurodegenerative diseases are significant.

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