behavioralmouseob/ob obese mice and lean mice with diet-induced obesity
Objective: Measurement of body weight, fat mass, glucose, and insulin levels in mice after ghrelin administration under high fat diet, and assessment of effects on metabolic parameters and gene expression
Materials & Equipment Checklist
8 items
Gather these items before starting the experiment. Check off items as you prepare.
Equipment4
Not specified • Not specified • Not specified • Not specified
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Protocol Steps
View Abstract
Background and aims: Ghrelin, an endogenous ligand for growth hormone secretagogue receptor (GHS-R), is an appetite stimulatory signal from the stomach with structural resemblance to motilin. We examined the effects of the gastric peptide ghrelin and GHS-R antagonists on energy balance and glycaemic control in mice. Materials and methods: Body weight, fat mass, glucose, insulin, and gene expression of leptin, adiponectin, and resistin in white adipose tissue (WAT) were measured after repeated administrations of ghrelin under a high fat diet. Gastric ghrelin gene expression was assessed by northern blot analysis. Energy intake and gastric emptying were measured after administration of GHS-R antagonists. Repeated administration of GHS-R antagonist was continued for six days in ob/ob obese mice. Results: Ghrelin induced remarkable adiposity and worsened glycaemic control under a high fat diet. Pair feeding inhibited this effect. Ghrelin elevated leptin mRNA expression and reduced resistin mRNA expression. Gastric ghrelin mRNA expression during fasting was increased by a high fat diet. GHS-R antagonists decreased energy intake in lean mice, in mice with diet induced obesity, and in ob/ob obese mice; it also reduced the rate of gastric emptying. Repeated administration of GHS-R antagonist decreased body weight gain and improved glycaemic control in ob/ob obese mice. Conclusions: Ghrelin appears to be closely related to excess weight gain, adiposity, and insulin resistance, particularly under a high fat diet and in the dynamic stage. Gastric peptide ghrelin and GHS-R may be promising therapeutic targets not only for anorexia-cachexia but also for obesity and type 2 diabetes, which are becoming increasingly prevalent worldwide.
1
Baseline measurements
Measure body weight, fat mass, glucose, and insulin levels in mice prior to treatment
Not specifiedNot specified
Note: Baseline values needed for comparison with post-treatment measurements
View evidence from paper
“Body weight, fat mass, glucose, insulin measured after repeated administrations of ghrelin”
2
Ghrelin administration under high fat diet
Administer ghrelin repeatedly to mice maintained on a high fat diet
Not specifiedNot specified
Note: Repeated administrations performed; specific dosage and frequency not stated
View evidence from paper
“Body weight, fat mass, glucose, insulin measured after repeated administrations of ghrelin under a high fat diet”
3
Post-treatment metabolic measurements
Measure body weight, fat mass, glucose, and insulin levels after ghrelin administration
Not specifiedNot specified
Note: Same parameters measured as baseline for comparison
View evidence from paper
“Body weight, fat mass, glucose, insulin measured after repeated administrations of ghrelin under a high fat diet”
4
White adipose tissue collection and gene expression analysis
Collect white adipose tissue and measure gene expression of leptin, adiponectin, and resistin
Not specifiedNot specified
Note: Gene expression measured in white adipose tissue
View evidence from paper
“Gene expression of leptin, adiponectin, and resistin in white adipose tissue were measured”
5
Gastric ghrelin gene expression assessment
Assess gastric ghrelin gene expression by northern blot analysis
Not specifiedNot specified
Note: Measurements taken during fasting state
View evidence from paper
“Gastric ghrelin gene expression was assessed by northern blot analysis”
6
GHS-R antagonist administration in ob/ob mice
Administer GHS-R antagonist repeatedly to ob/ob obese mice for six days
Six daysNot specified
Note: Repeated administration protocol; specific dosage not stated
View evidence from paper
“Repeated administration of GHS-R antagonist was continued for six days in ob/ob obese mice”
7
Energy intake measurement after GHS-R antagonist
Measure energy intake in lean mice, diet-induced obese mice, and ob/ob obese mice after GHS-R antagonist administration
Not specifiedNot specified
Note: Measured in three different mouse populations
View evidence from paper
“Energy intake and gastric emptying were measured after administration of GHS-R antagonists”
8
Gastric emptying measurement
Measure the rate of gastric emptying after GHS-R antagonist administration
Not specifiedNot specified
Note: GHS-R antagonists reduced the rate of gastric emptying
View evidence from paper
“Energy intake and gastric emptying were measured after administration of GHS-R antagonists”
9
Final body weight and glycaemic control assessment
Measure body weight gain and glycaemic control in ob/ob obese mice after six days of GHS-R antagonist treatment
Not specifiedNot specified
Note: Repeated administration of GHS-R antagonist decreased body weight gain and improved glycaemic control
View evidence from paper
“Repeated administration of GHS-R antagonist decreased body weight gain and improved glycaemic control in ob/ob obese mice”
Subjects / Specimens
Species
mouse
Strain
ob/ob obese mice and lean mice with diet-induced obesity
Age
Not specified
Sex
unknown
Weight
Not specified
Study included ob/ob obese mice and lean mice with diet-induced obesity