DISCUSSION

For the fed rat the glucose concentration in the heart tissue was 6.74µmol per gram and
the glycogen content was 0.109g per 100g of tissue. The heart tissue of the fasted rat had
a glucose concentration of 0.8709 µmol per gram and a glycogen content of 0.0141g per
100g. In the fed heart tissue, there is a higher content of glucose and glycogen when
compared to that of the fasted rat. This occurred since the fed rat had more glucose
available because of carbohydrate consumption from its diet. The little amount of stored
glycogen in the heart was broken down to supply the heart with glucose as an energy
source for aerobic respiration of the heart tissue.
For the kidney tissue of the fed rat, the glucose content was found to be 15.97µmol and
the glycogen content was 0.259 g per 100g of tissue. The fasted rat had a glucose content
of 0.4125µmol per gram of kidney tissue and a glycogen content of 0.00668g per 100g of
tissue. The glucose and glycogen content of the fed rat’s kidney was higher than that of
the fasted rat. This occurred since the kidney uses glucose from the bloodstream and does
not store glycogen actively. The kidney of the fasted rat had lower contents of glucose
and glycogen since glycogenolysis broke down the small amounts of glycogen present to
produce glucose so that the organs can be supplied with energy. Because of this the
content of glycogen present was depleted whilst the glucose content was temporarily
increased and then decreased due to disintegration of produce ATP for metabolism. The
glucose being used as an energy source in the fasted rat is not being reabsorbed in the
bloodstream to maintain an equilibrium since it is not receiving glucose from its diet thus
lowering its blood-glucose level. But since the fed rat is receiving glucose in its diet, an
equilibrium is maintained between energy utilization and glucose reabsorption in the
bloodstream increasing its blood glucose level causing its glucose and glycogen content
to be higher.
For the muscle tissue of the fed rat, the glucose content was found to be 24.975µmol and
the glycogen content was 0.405 g per 100g of tissue. The fasted rat had a glucose content
of 2.5 µmol per gram of muscle tissue and glycogen content of 0.0405g per 100g of
tissue. In the muscle the presence of the glucose-6-phosphatase enzyme causes the
hydrolysis of glucose-6-phosphate to form free glucose and a phosphate group. This
reaction occurs in the last step in gluconeogenesis and glycogenolysis. Because of this
glycogen cannot be stored in the muscle. When the enzyme is absent, glucose cannot
enter the blood steam. To counteract this, the glucose is stored as glycogen which is used
for the contraction of skeletal muscle tissues. When the glycogen is depleted it is not
replenished. Because of this the muscle tissues of the fasted rat had a lower glycogen and
glucose content than the fed rat. In the fed rat, the consumption of food replenished the
glycogen in the muscle tissues.
For the liver tissue of the fed rat, the glucose content was found to be 162.4µmol and the
glycogen content was 2.63 g per 100g of tissue. The fasted rat had a glucose content of
0.5465 µmol per gram of liver tissue and a glycogen content of 0.00885 g per 100g of
tissue. The glucose and glycogen contents of the fed rat was significantly higher than that
of the fasted rat. The higher glucose and glycogen contents in the liver tissue of the fed
rat was due to the increase in blood glucose levels after a meal. The excess molecules of
glucose are joined into branching chains of glycogen by gluconeogenesis through
condensation reactions. The liver functions to decrease the blood glucose levels and
increase the glycogen level. Consequently, during fasting when glucose molecules are
used to supply energy via glycolysis, the level of glucose in the liver is depleted. As a
result glycogen breakdown by glycogenolysis is stimulated by glucagon and the glucose
formed as a result is deposited into the bloodstream (Hames and Hooper 2005). Due to
this in the fasted rat, glycogen and glucose content of the liver is lower since the rat is not
consuming any form of glucose.
From the experiment it was observed that the tissues of the fed rats had higher glucose
content than glycogen. Glycogen is a storage form of glucose when it is present in
abundance and since the fed rats were being provided with a form of glucose, the tissues
obtained glucose in abundance and facilitated the storage of excess. In the fasted rat
tissues, the glucose content was also noticed to be higher than the glycogen content. This
occurred since no form of glucose was not provided for the fasted rats and hence glucose
was being utilised instead of being stored as glycogen. Both glucose and glycogen
content was higher in the fed rat tissues since these rats were being provided with a form
of glucose in the diet to replenish what was being used up by the tissues. The fasted rats
were not provided with any form of glucose and hence the glycogen and glucose contents
were lower since what was being used was not being replenished.
Under fed and fasted states the heart, muscle, kidney and liver tissues uses glycolysis,
gluconeogenesis,glycogenesis and glycogenolysis in different ways. This was
demonstrated by the results since the heart contained the least amount of glucose and
glucagon whilst the liver contained the most in the fed state. In the fasted state the kidney
had the least content of glycogen and glucose whilst the muscle had the most
The fed heart had the lowest glucose and glycogen content that all four of the tissues
since it respires constantly using the glucose that is available to it and converts its
glycogen stores into glucose to provide energy via glycolysis as ATP for the continuous
contraction and relaxation of the cardiac muscles. The fed kidney had a higher glycogen
and glucose content than the fed heart. This occurred since the kidney uses glucose and
absorbs it back from the bloodstream. The muscle tissues of the fed rat has a large
glucose and glycogen content than the fed kidney tissues. The fed liver tissue had a
higher glucose and glycogen content than the liver and also the highest of all the four fed
tissues. This occurred since the cells of the muscle has a lower glycogen content per
tissue gram since unlike the liver cells they do not store large amounts of
glycogen.Glucagon acts on the liver to break down glycogen to glucose and in the
muscle, the absence of the enzyme glucose-6-phosphatase allows for the
glucose-6-phosphate to remain in the tissue and to be subjected to glycolysis to facilitate
the muscle contraction due to movement,hence the glucose content in muscle tissue is
less than that in the liver.
In the fasted tissues the kidney had the lowest glycogen and glucose content since during
periods of fasting, the level of blood glucose becomes depleted as it is not supplemented
by the addition of glucose from meal consumption. There would be less glucose available
in the bloodstream, the kidney would have access to less glucose and the little amount of
stored glycogen would be converted into glucose to supply energy without being
replenished.
In the fasted liver tissue of the rat there was a higher glucose and glycogen content than
the fasted kidney but a lower content than the heart. This occured since the liver acts as a
store for glycogen. This store was depleted without being replenished since the body
could not store glycogen since glucose was required in various tissues,
The fasted heart tissue had a higher glucose and glycogen content than the liver. The
heart requires a constant energy supply inorder to facilitate constant contraction and
relaxation. Since the heart does not store glycogen, it is broken down into glucose which
is then used to produce 2ATP molecules to provide energy.
The fasted muscle tissue had the highest glycogen and glucose content out of all of the
fasted tissues. This occurred since the muscle had an ample amount of glucose present for
energy metabolism.

PRECAUTIONS
Foil was used to protect the light sensitive complex formed.
The pellet was properly dissolved to ensure that almost all of the glycogen was dissolved.
SOURCES OF ERRORS
The assay was sensitive to other compounds that were present such as lipids
The enzyme cocktail used was light sensitive.
ADDITIONAL DISCUSSION
In the liver glycogen acts as a reserve supply of glucose in order to keep the blood
glucose concentration at an adequate level throughout the body and to supply the brain
with fuel during periods of starvation, fasting or an increase in glucose usage due to
physical activity. Hence carbohydrates consumed are stored as liver glycogen, and during
fasting the glycogen is broken down and glucose is released into the blood.
In the heart tissues, very small amounts of glycogen are stored as it utilizes glucose,
ketones and mainly fatty acids as its energy source. When fatty acid, ketones and glucose
levels are dangerously low, the stored glycogen is broken down to glucose as an alternate
process to obtain energy by glycolysis. This energy is made available slowly to the heart
tissue.
In the muscle glycogen provides the necessary fuel for the muscle contraction during
physical activity. The contraction of the muscles during physical activity stimulated for
the breakdown of glycogen indirectly. During strenuous activity anaerobic metabolism
occurs causing the skeletal muscles to produce lactic acid from glycogen since oxygen is
not being supplied at a quick rate. The lactic acid can pass through the blood and become
taken up by the liver which converts it into glucose. The glucose produced is used to raise
the blood glucose. This is an indirect method to raise blood glucose levels since muscle
glycogen cannot be broken down to glucose and so cannot be used to raise blood glucose
concentration directly.
In the kidney tissues, glucose is taken up from the blood steam and stored at glycogen
Because of this there are little amounts of glycogen present in the kidney. The glycogen
in the kidney is broken down into glucose during fasting in order to achieve energy in the
form of ATP (a net gain of 2 ATP per glucose molecule) for metabolic reactions and
processes within the liver.
Glucose is stored as glycogen in large cytosolic granules. This is done since liver cells,
also known as hepatocytes, store glycogen equivalent to a glucose concentration of 0.4M.
The actual concentration of glycogen, which is insoluble and contributes little to the
osmolarity of the cytosol, is about 0.01 M. If the cytosol contained 0.4 M glucose, the
osmolarity would be threateningly elevated, leading to osmotic entry of water that might
rupture the cell
In humans most of the energy is stored as fats. This is done since fats provide 9 calories
per gram of fat, compared with 4.5 calories per gram of protein or carbohydrate. Fats can
also be stored anhydrous therefore it would occupy less storage space when storing when
compared to carbohydrates which require water for storage. (Cox 2013)
 NAME: Tishelle McCarthy

ID: 816008261

LAB PARTNER: Isha Khan

DEMONSTRATOR: Amanda

LAB # 5

LAB TITLE: Assay Of Tissue Glycogen