Feeding Behavior Assay
Objective: Measurement of food intake following direct VTA administration of ghrelin or GHSR antagonist to determine the role of ghrelin signaling in the ventral tegmental area on feeding behavior
Gather these items before starting the experiment. Check off items as you prepare.
Equipment2
Materials3
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Protocol Steps
Surgical preparation for VTA injection
Prepare animals for stereotaxic injection into the ventral tegmental area
Note: Procedure performed in mice and rats
View evidence from paper
“Direct VTA administration of ghrelin also triggered feeding, while intra-VTA delivery of a selective GHSR antagonist”
Administer ghrelin to VTA
Inject ghrelin directly into the ventral tegmental area of experimental animals
Note: Direct VTA administration triggers feeding behavior
View evidence from paper
“Direct VTA administration of ghrelin also triggered feeding”
Measure food intake following ghrelin administration
Quantify the amount of food consumed by animals following VTA ghrelin injection
Note: Food intake is measured as outcome of ghrelin signaling in VTA
View evidence from paper
“Direct VTA administration of ghrelin also triggered feeding”
Administer GHSR antagonist to VTA
Inject selective GHSR antagonist directly into the ventral tegmental area
Note: Antagonist blocks orexigenic effects of circulating ghrelin
View evidence from paper
“intra-VTA delivery of a selective GHSR antagonist blocked the orexigenic effect of circulating ghrelin”
Measure food intake following GHSR antagonist administration
Quantify food consumption following VTA GHSR antagonist injection
Note: Measures ability of antagonist to block feeding responses
View evidence from paper
“intra-VTA delivery of a selective GHSR antagonist blocked the orexigenic effect of circulating ghrelin and blunted rebound feeding following fasting”
Test rebound feeding following fasting
Measure food intake in animals subjected to fasting periods after GHSR antagonist administration
Note: GHSR antagonist blunts rebound feeding response after fasting
View evidence from paper
“intra-VTA delivery of a selective GHSR antagonist blocked the orexigenic effect of circulating ghrelin and blunted rebound feeding following fasting”
Test feeding responses in ghrelin-deficient mice
Measure feeding behavior in ghrelin-deficient mice under restricted feeding schedules
Note: Ghrelin-deficient mice show attenuated feeding responses to restricted feeding
View evidence from paper
“ghrelin- and GHSR-deficient mice showed attenuated feeding responses to restricted feeding schedules”
Test feeding responses in GHSR-deficient mice
Measure feeding behavior in GHSR-deficient mice under restricted feeding schedules
Note: GHSR-deficient mice show attenuated feeding responses to restricted feeding
View evidence from paper
“ghrelin- and GHSR-deficient mice showed attenuated feeding responses to restricted feeding schedules”