Abstract
Extrinsic control of single neurons and neuronal populations is a powerful approach for understanding how neural circuits function. Adding new thermogenetic tools to existing optogenetic and other forms of intervention will increase the complexity of questions that can be addressed. A good candidate for developing new thermogenetic tools is the Drosophila gustatory receptor family, which has been implicated in high-temperature avoidance behavior. We examined the five members of the Gr28b gene cluster for temperature-dependent properties via three approaches: biophysical characterization in Xenopus oocytes, functional calcium imaging in Drosophila motor neurons, and behavioral assays in adult Drosophila. Our results show that Gr28bD expression in Xenopus oocytes produces a non-specific cationic current that is activated by elevated temperatures. This current is non-inactivating and non-voltage dependent. When expressed in Drosophila motor neurons, Gr28bD can be used to change the firing pattern of individual cells in a temperature-dependent fashion. Finally, we show that pan-neuronal or motor neuron expression of Gr28bD can be used to alter fruit fly behavior with elevated temperatures. Together, these results validate the potential of the Gr28bD gene as a founding member of a new class of thermogenetic tools.
Highlights
Extrinsic control of cellular activity is a powerful paradigm for understanding how neural circuits regulate behavior
We examined the temperature dependence, ionic selectivity, and basic kinetic properties of Gr28bD, using the two-electrode voltage-clamp (TEVC) technique to record oocyte currents under different conditions of temperature, membrane potential, and ionic composition of the extracellular solution
The development of thermogenetic tools as a complement to optogenetics and other approaches is a promising step toward multi-modal control of neuronal activity[1,2]
Summary
Extrinsic control of cellular activity is a powerful paradigm for understanding how neural circuits regulate behavior. With a response threshold of about 25 °C and abrupt activation with temperature[6,10], the TRP protein family (e.g., TRPA1) has been the most extensively studied and applied to understand neural circuits in Drosophila and other organisms[11,12,13]. Their potential for thermogenetic applications has not been explicitly tested so far, the Drosophila Grs are promising new candidates. These results validate the potential of the Gr28bD gene as a founding member of a new class of thermogenetic tools
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