Feeding behavior assay
Objective: Measurement of food intake and feeding behavior in response to AgRP neuron activation using DREADD technology to demonstrate that AgRP neuron activity is necessary and sufficient for feeding
Gather these items before starting the experiment. Check off items as you prepare.
Equipment2
Materials1
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Protocol Steps
AgRP neuron activation via DREADD
Activate AgRP neurons using DREADD technology with designer drug administration to provide specific and reversible regulation of neuronal activity
Note: DREADD technology allows for acute and chronic regulation of AgRP neurons
View evidence from paper
“Using designer receptors exclusively activated by designer drugs (DREADD) technology to provide specific and reversible regulation of neuronal activity in mice”
Measure food intake during activation
Quantify food consumption in response to acute AgRP neuron activation
Note: Activation rapidly and dramatically induces feeding behavior
View evidence from paper
“acute activation of AgRP neurons rapidly and dramatically induces feeding”
Measure energy expenditure during activation
Quantify metabolic rate and energy expenditure during AgRP neuron activation
Note: Energy expenditure is reduced during AgRP neuron activation
View evidence from paper
“acute activation of AgRP neurons rapidly and dramatically induces feeding, reduces energy expenditure”
Monitor fat storage changes
Measure changes in fat stores following AgRP neuron activation
Note: Fat stores increase as a result of AgRP neuron activation
View evidence from paper
“acute activation of AgRP neurons rapidly and dramatically induces feeding, reduces energy expenditure, and ultimately increases fat stores”
Withdraw stimulation and monitor recovery
Remove DREADD activation and measure return of all parameters to baseline
Note: All effects return to baseline after stimulation withdrawal, demonstrating reversibility
View evidence from paper
“All these effects returned to baseline after stimulation was withdrawn”
AgRP neuron inhibition in hungry mice
Inhibit AgRP neuronal activity in hungry mice and measure food intake reduction
Note: Demonstrates that AgRP neuron activity is necessary for feeding
View evidence from paper
“inhibiting AgRP neuronal activity in hungry mice reduced food intake”
Measure feeding motivation and food-seeking behavior
Assess motivation for feeding and food-seeking behavior during AgRP neuron activation
Note: AgRP neuron activation potently increases motivation and drives intense food-seeking behavior
View evidence from paper
“activating AgRP neurons potently increased motivation for feeding and also drove intense food-seeking behavior”