Veterinary Physiology 143

Laboratory Exercise No. 10 Insulin Induced Convulsion

Section: A-3L Mon 9-12

Group:
Chu
Manalo
Masajo
Introduction

Glucose is the primary fuel of the animals body in order for it to function
normally and efficiently thus decrease in availability of these molecules will lead to
altered physiological state of different organs utilizing it. The body would want to
regulate and maintain blood glucose in a normal narrow range. Thus, insulin and
glucagon are produced.
Like all pancreatic hormones, Insulin is a peptide hormone significantly
produced by cells of the Islet of Langerhans. While, Glucagon is secreted by the cells of the pancreatic islet of Langerhans of the pancreas in which these cells sense
the glucose level in blood and detect if theres abnormality. They are both secreted
in response to blood sugar levels, but have opposite physiologic activities. Insulin is
responsible for promoting glucose uptake from the blood and into skeletal muscle
and fat tissue, consequently decreasing blood glucose level. On the other hand, the
effect of glucagon is to make the liver release the glucose it has stored in its cells
into the bloodstream, with the net effect of increasing blood glucose. Glucagon also
induces the liver (and some other cells such as muscle) to make glucose out of
building blocks obtained from other nutrients found in the body (eg, protein).
For this exercise, the main hormone in focus is Insulin. Insulin is secreted by
the body in constant regulated proportions. Secretions can be stimulated by high
glucose level or hyperglycemia brought about by physiologic factors, hormones,
intestinal hormones and drugs or substances. Insulin secretion is inhibited by low
glucose level or hypoglycemia which could be induced by hormones or drugs. The
excessive iatrogenic induction of insulin would cause hyper-insulinism. This would
elicit lowering of blood glucose level or hypoglycemia and utilization of body fats as
energy source or lipolysis. Hyperinsulinemic hypoglycemia can cause shakiness,
weakness, seizures, coma and death. In order to recuperate the effects of
hyperinsulinism, we must test for the effect of glucose and epinephrine.

Objectives
The objective of this experiment is to demonstrate the effect of iatrogenic
hyperinsulinism and to compare the effectiveness of glucose and epinephrine in the
treatment of insulin-induced hypoglycemia.

Materials and Methods

Distilled water
Tuberculin syringe
50% sterile glucose solution

regular insulin
epinephrine
gavage needle

Glucometer
4 adult laboratory mice (same sex and weight)

Label mice A, B, C and D.

A

Fast for 24
hours

Collect blood via tail v.

Measure blood glucose
Inject 4 units
levelof insulin
Observe closely and note changes in RR and
SC
Note
time onset of tremors and convulsions
behavior
Collect blood
Measure blood glucose
Give the following
level
Epinephrine 0.1mL
Epinephrine 0.1mL
treatment:
C
A
(IV)
(IV)
Glucose (gavage)
Glucose (gavage)
D
B
Observe and note time tremors and convulsion stops. Collect blood and
measure blood glucose level

Results
The following results were acquired after performing the experiment.
Table1. Observed changes in RR after SC injection of Insulin
Anim
RR before insulin
RR after insulin injection
al
injection
A
28
144
B
32
128
C
32
132
D
29
120
There is a noticeable increase in respiration rate. This is due to the activation of
glycolytic process due to insulin. The basal metabolic rate (BMR) of cells increases
and the speed of oxygen intake also increases to compensate the CO 2 rapidly being
produced with increases metabolism.
Convulsions were observed after insulin injection. The reason as how convulsions
happen is explained by this diagram:

Hypoglycemia
(low blood glucose
level)

Abnormal
uncontrolled
neuronal

Respirati
on

Na/K
ATPase

(reduced)

(imparement)

Interrupted
neuronal
electrical flow in
the brain

LOW ATP
producti
on
Silent/ Nonfunctional
neurons

Spread
ofutilizes glucoseGeneralized
Take note that
the brain
as a source of energy. It is only expected
brain
discharge
that convulsion
will occur since.
stimulation

Table2. Blood glucose level (mmol/L) after SC injection of Insulin

Anim
al

Just before insulin

injection

Time when tremors &

convulsions was observed

CONVULSI
ON

Time when tremors &

convulsions ceased after
administration of glucose
or epinephrine

A
5.0
LO*
B
3.7
1.2
1.2
C
10.3
LO
3.9
D
4.9
1.4
4.9
*LO on glucometer meant a blood glucose concentration <1.1 mmol/L)
Table3. Time when tremors and convulsions are observed and time when tremors
and convulsions ceased after administration of glucose or epinephrine
Anim
al

Time when tremors &

convulsions observed

A
B
C
D

12:35 PM (2h 15 min)

10:55 AM (35 min)
1:16 PM (2h 56 min)
11:08 AM (43 min)

Time when tremors & convulsions

ceased after adm of glucose or
epinephrine
12:45 PM (10 min)
11:00 AM (5 min Time of death)
1:32 PM (16 min)
11:10 AM (2 mins)

It was observed that the mice with the lowest blood glucose concentration had
convulsions the earliest. Ideally, the fasted mice had to have the lowest blood
glucose but due to the possibility that mice A must have eaten more and mice D
had a low appetite can only explain the unexpected glucose level result.
On the other hand, mice B had the lowest blood glucose level and had convulsion in
the shortest time. It was not able to survive as its glucose reserve had been
depleted turning it unable to even process glucose given by gavage needle.
The action of insulin is to decrease blood glucose level. In the case of this study,
insulin administration further decreased the already depleted blood glucose
concentration that tremors occurred. Treatment of glucose and epinephrine acts to
replenish blood glucose levels where in the mice is no more in a life threatening
situation seen as the absence of convulsions.

Discussion
Insulin functions to decrease the blood glucose level. Insulin signals the cells of the
body to take in glucose, decreasing blood glucose levels, storing it in cells and

tissues. This explains the decrease in glucose in circulating blood. It is no longer

present in blood but in cells and tissues of the body.
Ideally, fasted animals have a lower blood glucose levels in contrast with the fed
animal. After insulin injection, blood glucose decreases and once convulsion occur,
glucose levels are at its fatal level enough that neurologic sign is observed. Once
the treatment is given, blood glucose level starts to elevate as treatments of
glucose and epinephrine alters insulin action which will be further explained on the
following diagrams. Blood glucose is back to its baseline after the cessation of
tremors. This is because the depleted glucose had been replenished and that
neurologic signs are addressed.
Effect of Epinephrine on Hypoglycemic animal:

Insulininduced
hypoglycemia

Epinephrin
e injection

Epinephrine
in Blood

Increase in
Glucose Blood
level
Increase in:
Liver glucose output
Glucagon levels in
blood
Available
gluconeogenic
precursors

Epinephrine
bind to and
adrenergic
receptors

Glucose in
the Brain

Normalized
Electrolytes and
Glucose Level in
the Brain

A review on the physiology of digestion demonstrates the key players of metabolism

and regulation. Glucose uptake is favored by the cholinergic nervous system.
Administration of epinephrine would activate the adrenergic receptors and in turn
negate the action of insulin. Action of epinephrine depends on the type adrenergic
receptor in which it will bind and the expression of G-protein molecules by the cells
due to epinephrine-receptor stimulation.
We have learned that there are two types of adrenergic receptors (1, 2 and ).
Binding of epinephrine to 1-adrenergic result in the expression of G-protein (Gg)
which in turn activates phospholipase C, resulting to the production of inositol
triphosphate and diacylglycerol. The produced inositol triphosphate causes the
release of Ca3+ from the ER to stimulate liver output of glucose and skeletal muscle

production of lactate, a precursor of glucose in glucose synthesis. Binding of

epinephrine to 2-adrenergic receptor leads to a decrease synthesis of cAMP.
Lastly, binding of epinephrine to adrenergic receptors activates adenylyl cyclase
which results to an increase in cAMP and later on activates PKA. Activation of PKA is
important in increasing glucagon levels, promoting glycogenolysis and
gluconeogenesis. Glycolysis in skeletal muscles increases the availability of
important glucose synthesis precursors such as alanine and lactate. These are later
processed by the liver and kidneys.

Effect of Glucose on Hypoglycemic animal in a diagram:

Treatment
with Glucose
(gavage)

Glucose uptake
by the intestinal
cells

Increased
Intravascular
Glucose
concentration

Glucose in
interstitium

Glucose
transported by
GLUT-1 across
BBB

Sufficient
Glucose
concentration in
the Brain

Normalization of
Electrolytes and
EnergySubstrate Level
in the Brain

Caesation of
Convulsion

Induced
Hypoglycemia

For the blood glucose to increase from glucose solution administered using a
gavage needle, glucose must first be absorbed by intestinal epithelial cells of the
body into the blood stream. After which, glucose is then distributed throughout the
body and to the brain via glucose transporter 1(GLUT-1), where in it is able to cross
the BBB and supply the brain.
Reviewing the results shown at table 3, the glucose treatment was faster in
relieving tremors by hyperinsulinism. This is due to a faster increase of blood
glucose level and a less number of precursor steps in glucose synthesis.

Conclusion
Hyperinsulinism, which induces very low blood glucose level in an animals
body, can lead to occurrence of tremors and convulsion and thus the animals
undergoing insulin treatment must be closely monitored in order to prevent
undesirable effects or even death. Glucose administration would be a better choice
than epinephrine if one would want to alter the effect of hyperinsulinism because
glucose administration would lead to faster reaction.

References

Guyton Hall, Textbook of Medical Physiology. 11th Edition. Pp. 939-954

Arieff,A.I.,Doerner,T.,Zelig,H. & Massry,S.G. (1974). Mechanisms of Seizure

and Coma in Hypoglycemia:EVIDENCE FOR A DIRECT EFFECT OF INSULIN
IN ELECTROLYTE TRANSPORT IN BRAIN.The Journal of Clinical
Investigation.pp. 654-661. Retrieved on August 30,2012 from
http://www.jci.org/articles/view/107803

Bowen, R. 2009. Insulin Synthesis and Secretion. Retrieved on November

16, 2014 from
http://www.vivo.colostate.edu/hbooks/pathphys/endocrine/pancreas/insuli
n_phys.html

Sinha, Sunil. 2014. Hyperinsulinism. Rretrieved on November 16, 2014

from
http://emedicine.medscape.com/article/921258-overview