Anthrone Colorimetric Method for Quantifying Glycogen in Rat Liver and Skeletal Muscle

Area of Science:

• Biochemistry
• Physiology
• Metabolic Research

Background:

• Glycogen serves as a crucial glucose reserve in liver and muscle.
• Hepatic glycogen maintains blood glucose levels, while muscle glycogen fuels physical activity.
• Accurate glycogen quantification is vital for studying metabolic regulation and disease.

Purpose of the Study:

• To develop and validate a sensitive anthrone colorimetric method for quantifying tissue glycogen.
• To assess glycogen levels in rat liver and skeletal muscle under diverse physiological conditions.
• To provide a reliable tool for in vivo and in vitro glycogen metabolism research.

Main Methods:

• Alkaline digestion, deproteinization, glycogen precipitation, and alcohol washing.
• Colorimetric quantification using anthrone reagent.
• Orthogonal validation with western blot analysis of glycogen synthase and phosphorylase.

Main Results:

• The anthrone method reliably quantifies glycogen in rat liver and muscle.
• The method demonstrates sensitivity across fed, fasted, and refed states.
• Epinephrine-induced glycogenolysis was successfully modeled, showing method's dynamic range.

Conclusions:

• The described anthrone colorimetric method is a sensitive and reliable tool for glycogen quantification.
• This method is applicable to various physiological states and stress conditions.
• It facilitates research into carbohydrate metabolism, both in vivo and in vitro.