Cold-induced hyperphagia requires AgRP-neuron activation in mice
Cite this dataset
Deem, Jennifer (2020). Cold-induced hyperphagia requires AgRP-neuron activation in mice [Dataset]. Dryad. https://doi.org/10.5061/dryad.0p2ngf208
To maintain energy homeostasis during cold exposure, the increased energy demands of thermogenesis must be counterbalanced by increased energy intake. To investigate the neurobiological mechanisms underlying this cold-induced hyperphagia, we asked whether agouti-related peptide (AgRP) neurons are activated when animals are placed in a cold environment and, if so, whether this response is required for the associated hyperphagia. We report that AgRP-neuron activation occurs rapidly upon acute cold exposure, as do increases of both energy expenditure and energy intake, suggesting the mere perception of cold is sufficient to engage each of these responses. We further report that silencing of AgRP neurons selectively blocks the effect of cold exposure to increase food intake but has no effect on energy expenditure. Together, these findings establish a physiologically important role for AgRP neurons in the hyperphagic response to cold exposure.
Mice expressing GCaMP6s in AgRP neurons were connected to a fiber-photometry system to enable fluorometric analysis of real-time neuronal activity. Briefly, for calcium recording in vivo, two excitation wavelengths (470 nm and 405 nm isosbestic) were used to indicate calcium-dependent and calcium-independent (i.e., due to bleaching and motion artifacts) GCaMP6s fluorescence, respectively. Light was delivered via fiber-coupled LEDs (LED lights: M470F3 and M405FP1, LED driver: DC4104, Thorlabs, Newton, NJ) and modulated by a real-time amplifier (RZ5P, Tucker-Davis Technology (TDT), Alachua, FL) at non-divisible frequencies (331 Hz and 231 Hz, respectively) to prevent signal interference between the channels. Excitation lights were bandpass filtered (475 ± 15nm , 405 ± 5nm; iFMC4, Doric Lenses, Quebec, QC, Canada) and the combined excitation light delivered through a fiberoptic patch cord (M75L01,Thorlabs) connected to a rotary joint (FRJ, 0.48 NA, Ø400μ m core; Doric Lenses) to prevent fiberoptic torsion during animal movement. A final connector patch cord (MFP, 0.48 NA, Ø400 μ m core; Doric Lenses) was connected to the implanted fiberoptic via a ceramic mating sleeve (ADAL1, Thorlabs). Emitted light was collected through the same patch cord, bandpass filtered (525 ± 25 nm; iFMC4) and transduced to digital signals by an integrated photodetector head. Electrical signals were sampled at a rate of 1017.25 Hz and demodulated by the RZ5P real-time processor. Experiments were controlled by Synapse software (TDT).
Custom MATLAB scripts were developed for analyzing fiber photometry data. The isosbestic 405-nm excitation control signal was subtracted from the 470-nm excitation signal to remove movement artifacts from intracellular calcium dependent GCaMP6s fluorescence. Baseline drift was evident in the signal due to slow photobleaching artifacts, particularly during the first several minutes of each recording session. A double exponential curve was fit to the raw trace of temperature-ramping experiments while a linear fit was applied to the raw trace of food presentation experiments and subtracted to correct for baseline drift. After baseline correction, dF/F (%) was calculated as individual fluorescence intensity measurements relative to median fluorescence of entire session for 470nm channel. Averaged dF/F (%) for each temperature (30 or 14°C) were limited to the 10-min period either before or after the 1-min ramp either between the 14°C to 30°C transition or the 30°C to 14°C transition.
Data is provided in .mat format.
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