Lecture 7

Topics covered:
- Puberty
- Endocrinology of pregnancy
- Pancreatic hormone
- Insulin Fed state
- Glucagon Fasting state
- Stimuli for secretion
- Receptor transduction mechanism
- Actions
- Hypo and hyper glycemia
- Diabetes mellitus and metabolic syndrome
Puberty:
Puberty begins with the beginning of the release of GnRH which stimulates the sexual organs.
The age of the onset of puberty is decreasing and is attributed to better nutrition. Secondary sexual
characteristics develop under the influence of sex steroids during puberty in both males and females.
- Height spurt
- Penis growth
- Testicular growth
- Pubic hair and axillary hair
- Increased muscle and lean mass in males
- Increased fat in females (it is relative to males)
- Body odor
- Menarche in females
- Breast size and changes in the body shape
- Lubrication of the vagina
- Sperm production
There has been a correlation between the body weight and the onset of puberty. Overweight kids tend
to undergo puberty earlier than lean children. This could relate to the storage of the sex hormones in
the adipose tissue present in the overweight people.
- Adipocytes secrete a hormone called Leptin which was discovered in rodents which were obese
and had mutation in the OB/OB gene. Leptos=thin
- OB/OB mutants did not undergo puberty and it is now believed that Leptin is involved in
triggering the signal for puberty.
- The size of the Adipocyte is related to the secretion of this hormone; bigger the size of
adipocytes, more the production and secretion of Leptin.
High levels of Leptin leads to increased appetite.

Fertilization:

Once the secondary oocyte is ovulated, it remains viable for fertilization for only about 24 hours.
Nevertheless, viable sperm may remain in the female reproductive tract for about 3-5 days after
ejaculation. It is hypothesized that the ovulated secondary oocyte may attract the sperm via chemotaxis.
Of the 100 million sperm ejaculated, only 100 reach the oocyte, and only one sperm will induce
fertilization. How is polyspermy prevented? In order to fertilize the egg, the sperm must penetrate
through the corona radiata, a layer of granulosa cells, as well as through the zona pellucida, a protective
mucopolysaccharide coat. In order to achieve this task, capacitated sperm undergo the acrosomal
reaction, which can only occur if the sperm undergo capacitation. The acrosomal reaction involves the
exocytosis of the acrosome, which contains enzymes that dissolve the corona radiata and the zona
pellucida, allowing the sperm to reach the egg and find the sperm binding receptors on the vitelline
layer, which is the outer layer of the ovum. The binding of the sperm to its receptor triggers fertilization
and the completion of Meiosis II by the ovum. Additionally, fertilization triggers the cortical reaction,
which is the main mammalian mechanism that safeguards against polyspermy. Before fertilization, the
egg plasma membrane is tightly connected to an outer membrane, called the vitelline layer. The egg
contains numerous vesicles filled with mucopolysaccharides, called cortical granules, along its periphery
in the cytosol. Upon sperm entry and fertilization, there is a sudden spike in calcium levels that starts at
the point of sperm entry and spreads through the entire surface of the egg. The increased calcium levels
trigger the exocytosis of the cortical granules into the intermembrane space between the vitelline layer
and the egg plasma membrane. The increased concentration of the cortical granules in the
intermembrane space results in a sharp osmotic gradient that serves to draw large volumes of water
into the intermembrane space, and thus, leads to the separation of the plasma membrane from the
vitelline layer, inhibiting sperm entry into the ovum.
Fertilization occurs in the ampulla, which is the portion of the oviduct closest to the fimbriae. The
embryo takes four to five days to traverse the oviduct and to reach the uterine cavity. About two days
after reaching the uterus and seven days after fertilization, the embryo implants in the endometrium.
During this time, the embryo undergoes rapid mitotic divisions so that the embryo exists as a blastula by
the time it reaches and implants in the uterus. The blastocyst consists of a hollow spherical structure
composed of approximately 100 cells. The trophoblast, or the cells in the outer layer of the blastocyst,
will form the chorion and placenta, whereas the cells of the inner cell mass will give rise to the fetus, the
amnion, which secretes amniotic fluid, and the allantois, which produces the umbilical cord.


Ectopic pregnancies refer to the implantation of the blastocyst in a site other than the uterus, for
example in the oviduct, or in the abdominal cavity external to the uterus. Ectopic pregnancies are very
dangerous and often fatal. A tubal pregnancy refers to the implantation of the blastocyst into the
oviduct. Because the embryo is the fastest growing tumor that grows at an exponential rate and the
oviduct cannot stretch like the uterus, a tubal pregnancy often results in fallopian tube rupture and
internal bleeding. The treatment for tubal pregnancy is the surgical removal of the affected oviduct; this
procedure does not render the patient infertile as the remaining oviduct may capture and propel an
ovulated secondary oocyte toward the uterus and result in conception.
The blastocyst secretes enzymes that break down and form a hole in the endometrium where the
embryo inserts itself. Soon after implantation, the endometrial layer grows back over the blastocyst, and
in doing so, completely covers the blastocyst within itself. As the blastocyst further grows and matures,
the chorion forms fingerlike projections called chorionic villi, which contain enzymes at their tips. The
chorionic villi further penetrate and breakdown the innermost layer of the endometrium, which is highly
vascularized. The breakdown of the vascularized endometrium forms pools of maternal blood that
surround the villi, which contain the embryonic blood vessels. The fetal circulation is structured in a
manner similar to that found in the pulmonary circulation in that the umbilical vein contains nutrients
and high oxygenated blood whereas the umbilical artery contains poorly oxygenated blood and
metabolic wastes. The chorionic villi are the sites of exchange between the maternal and fetal
circulations. Nutrients, gases, hormones, and antibodies are transferred from the maternal blood into
the fetal circulation via diffusion and facilitated transport, while fetal hormones and waste products
from the fetal circulation are transferred into the maternal circulation. Although the villi allow for the
exchange of nutrients and wastes, they do not allow for the mixing of maternal and fetal blood as the
villi membrane is impermeable to red blood cells.

Hormones during pregnancy
- hCG: Mainly substitutes for LH
- hPL: Stimulates the breast and also, more importantly, is similar to GH in its metabolic effect.
- Relaxin: it decreases the contractility of the uterus and is called so because it relaxes the pelvic
ligaments (connective tissue binding different bones together), leading to be able to have the
birth canal open up for the delivery of the baby.
- Fetus and mother both can provide androgens and estrogens (by conversion of these
androgens).
- 17-hydroxy progesterone: Secreted by the corpus luteum only and is used as a measure of
Corpus Luteum function by the physicians.
- Estrogen: Though the placenta can synthesize peptide hormones de novo, the placenta needs
steroid precursors in order to synthesize steroid hormones. Thus, the placenta uses DHEA-
Sulfate from the maternal and fetal adrenal cortex to synthesize estrogen. The placenta has a
sulfatase, which cleaves the sulfate from the DHEA, allowing DHEA to be converted to the
various estrogens.
Elevated levels of estrogen contribute to the development of the mammary glandular
structure as well as activating the nauseatory reflex of the medulla oblongata. Estrogen is also
believed to play a crucial role in the induction of labor. Estrogen promotes uterine contractility
by stimulating oxytocin upregulation on the myometrium thereby increasing the sensitivity of
the myometrium to oxytocin. Also, estrogen increases the number of gap junctions between the
myometrial cells. This latter effect results in consistent, unidirectional contractions that occur in
unison. Toward the end of the third trimester, the estrogen to progesterone ratio increases,
either by progesterone receptor downregulation, increased estrogen secretion, upregulation of
estrogen receptors, or a combination of these three effects.

- Thyroid Gland: Although TSH secretion does not increase during pregnancy, the production of
the thyroid hormones rises, especially during the first trimester. Human chorionic gonadotropin
acts as a weak TSH receptor agonist; thus, during the first trimester, when hCG levels are
highest, the mother may develop transient hyperthyroidism and the thyroid gland may undergo
slight hypertrophy. Specifically, early gestation is marked by increased T4 synthesis, whereas
late gestation is marked by increased T3 levels. Furthermore, increased estrogen levels
stimulate the increased synthesis of thyroglobulin, thus, increasing the half-life and serum levels
of the thyroid hormones.
- Adrenal Cortex: During pregnancy, the maternal adrenal cortex demonstrates increased activity
and hormone synthesis in all three zones. Glucocorticoid synthesis increases due to the
exceptionally high levels of pregnancy hormones, especially estrogen, which also stimulates
increased production of corticosteroid binding globulin, the carrier protein for cortisol. Thus,
both the synthesis and half-life of cortisol is increased during pregnancy. It is interesting to note
that pregnant women do not develop symptoms of Cushings disease and it has been suggested
that progesterone may be responsible for this effect by selectively antagonizing the action of the
corticosteroids. Thus, the main effect of cortisol during pregnancy is to increase insulin
resistance, in order to reserve glucose stores for the developing fetus. Also, increased cortisol
secretion may contribute to the development of striae, or stretch marks, commonly
associated with pregnancy.
Aldosterone secretion demonstrated eight times more secretion during pregnancy.
However, pregnant women do not commonly develop hypertension because progesterone is a
competitive inhibitor of the mineralcorticoid receptor, thus ensuring normal blood pressure
during pregnancy.
Androgen secretion, especially DHEA-Sulfate, increases during pregnancy in order to
support placental secretion of estrogens.
- Insulin: During gestational diabetes, high glucose will travels to the fetus and stimulates the
secretion of insulin. This can lead to the collection of water in the babies, leading to big size of
the babies.
- Oxytocin: By 12-18 weeks of gestation, the fetal posterior pituitary begins to secrete
vasopressin and oxytocin and the levels of both hormones progressively increase in the fetal
circulation throughout pregnancy. It has been suggested that fetal secretion of oxytocin may
contribute to the maintenance of labor because levels of oxytocin in the umbilical artery are
greater than oxytocin levels found in the fetal umbilical vein. In other words, the oxytocin levels
that the fetus secretes into the maternal circulation exceed the oxytocin levels the fetus
receives from the maternal circulation. Note, however, that this increase in oxytocin occurs only
after the induction of labor. Towards the end of the third trimester and prior to the induction of
labor, oxytocin levels are constant; however, increased estrogen action leads to an upregulation
of oxytocin receptors in the myometrium, increasing the effectiveness of oxytocin. It is only
after the fetus descends into the cervix, that the oxytocin positive feedback loop is triggered and
oxytocin levels rise exponentially. Thus, although fetal oxytocin probably does not induce labor,
it may play a significant role in maintaining labor by giving rise to consistent contractions.
Pancreas and its hormones
The pancreas is a dual function organ as it acts to maintain homeostatic control of digestion and
metabolism. About 98% of the pancreas is composed of exocrine cells, which secrete bicarbonate rich
juice and synthesize all of the digestive enzyme precursors. The remaining 2% of the total mass of the
pancreas is composed of small clusters of endocrine cells, called the islets of Langerhans that contain
three distinct cells types. About 75% of the islet of Langerhans are composed of beta cells, which secrete
insulin, 20% is composed of the alpha cells, which secrete glucagon, and about 4% are delta cells (or D
cells), which secrete somatostatin. Like all endocrine glands, the islets of Langerhans, are highly
vascularized and surrounded by capillary beds. In addition, they are closely associated with synapses of
autonomic neurons. Insulin and glucagon are antagonistic hormones in that insulin decreases blood
glucose levels whereas glucagon increases blood glucose. Somatostatin, which is also found in the
hypothalamus and stomach, plays a regulatory role by inhibiting glucagon and insulin secretion.
Moreover, these pancreatic peptide hormones, have a short half-life and they therefore must be
continuously secreted in order to exert sustained action. Importantly, because insulin and glucagon are
antagonistic hormones, it is the insulin to glucagon ratio that dictates the direction of metabolism.


Insulin: Insulin is a 51-amino acid peptide hormone that is secreted via the protein secretory pathway.
Insulin is synthesized as a single polypeptide chain that is cleaved upon package into a secretory vesicle
via protease action, which generates the connecting peptide (c-peptide) by-product and the active
insulin hormone with the alpha and beta fragments linked by disulfide bridges. The c-peptide, which
bears no known biological significance, is co-secreted with insulin in a 1:1 fashion and maintains a longer
half-life than insulin, which has a half-life of about five minutes due to its active metabolism by the liver.
Thus, clinicians often use c-peptide as a more accurate representation of endogenous insulin secretion
rates. The insulin receptor belongs to the class of growth factor receptors and contains an extracellular
alpha domain and a transmembrane beta domain that dimerize upon activation. It is secreted mainly in
the Fed state.
Stimuli for Insulin secretion:

1. Increased Plasma Glucose: This is the most potent stimulator of insulin secretion as it acts
directly on pancreatic beta cells to promote the exocytosis of insulin filled vesicles. Specifically,
when plasma glucose concentrations are greater than 100 mg/dL, insulin secretion dominates.
There are different glucose transporters:
a. GLUT1: Regular affinity and are present in most cells, including human beta cells. Fasting
glucose concentration is 70-115mg/dL. Below 70mg/dLhypoglycemia and over 125mg/dL.
b. GLUT2: is a very low affinity receptor and it will only bind and transport glucose when
concentration is high. It is found in the Kidneys; in the proximal tubules, small intestine and
liver cells (stores glucose in the form of glycogen). If there is not enough glucose, glycogen
can be broken down and released out of the liver to compensate for the decrease.
c. GLUT3: is mainly expressed in neurons and has much higher affinity than the first two.
When the glucose concentration begins to decrease and the neurons are the last ones to be
deficient of glucose.
d. GLUT4: are transporters that are made but are not inserted in the plasma membrane. They
exist in the secretory vesicles within the cells and upon certain signal, glucose gets released.
Insulin is the signal for secretion. These are found in 2 types of cells: skeletal muscle cells
and adipocytes. Exercise can increase the exocytosis of these GLUT4 transporters in the
muscles. These also have regular affinity for glucose.
2. Increased Plasma Amino Acids: The consumption of meals containing protein also promotes
insulin secretion.
3. GI Hormones: The entry of nutrients into the intestine triggers the secretion of a wide variety of
GI homones, and especially the secretion of one class of GI hormones called the incretins,
namely glucagon like peptide 1 (GLP-1) and gastric inhibitory peptide (GIP). These hormones
stimulate insulin secretion in a feed-forward manner, that is, the GI tract anticipates an increase
in plasma glucose and opts for the secretion of insulin before plasma glucose levels become too
high. This action prevents a sudden surge of plasma glucose and ensures homeostatic
maintenance during the digestion and absorption of nutrients. Notably, somatostatin inhibits
the secretion of these GI hormones and thus inhibits this feed-forward effect.
4. Parasympathetic Activity (Acetylcholine): Because the parasympathetic branch of the autonomic
nervous system dominates during situations of rest and digest, the ingestion of a meal
stimulates the vagus nerve, which contains neural circuits that innervate the pancreas directly,
thus stimulating insulin secretion.
5. Sympathetic Activity (Epinephrine, only): The stimulation of beta adrenergic receptors by
epinephrine also acts as a stimulator of insulin secretion. Beta adrenergic
receptorsstimulatory effects and Alpha 2 receptors Inhibitory effect (main stimulus).
6. Glucagon: Though glucagon is antagonistic to insulin, it weakly stimulates for insulin secretion
for two main reasons. First, the co-secretion of glucagon and insulin helps to fine-tune plasma
glucose levels in order to ensure that glucagon does not result in excess increase in plasma
glucose. The pancreatic alpha cells require insulin stimulation in order to take up glucose, which
inhibits glucagon secretion in a negative feedback manner. Insulin stimulation of alpha cells
results in the upregulation of glucose transporters (GLUT4) in the plasma membrane, which
allows for glucose entry into the cell. Thus, in order to maintain adequate levels of blood glucose
and to prevent glucagon-induced hyperglycemia, the pancreatic alpha cells need insulin in order
to take up glucose and to inhibit glucagon secretion as needed. Similarly, adipose tissue and
skeletal muscle, also require insulin stimulation in order to become capable of taking up glucose.
During times of stress or exercise, glucose needs to be transported to the active skeletal muscle
in order to maintain ATP production.
Mechanism of Insulin Secretion: Recall that increased plasma glucose levels stimulate insulin
secretion. Glucose diffuses into the pancreatic beta cell via the GLUT2 transporter when blood
glucose levels are high. The glucose is converted into ATP via oxidative phosphorylation. Pancreatic
beta cells contain special potassium leak channels that become blocked by ATP (KATP channel). Thus,
a high plasma glucose concentration translates into a high ATP concentration inside the pancreatic
beta cell, and thus, blockage of the KATP channel. When the KATP channel is not blocked by ATP, it
allows for potassium ions to diffuse out of the cell, maintain the hyperpolarized membrane
potential. However, when ATP blocks the KATP channel, potassium accumulates inside the pancreatic
beta cell, thus depolarizing the membrane potential (since potassium ions carry a positive charge).
This depolarization triggers the activation and opening of voltage gated calcium channels, thus
leading to net influx of calcium into the pancreatic beta cell. The increased cytosolic calcium levels
triggers the exocytosis of docked insulin filled vesicles, thus leading to increased plasma insulin
levels. However, measurement of insulin secretion rates shows that after reaching a maximal rate,
insulin secretion falls and remains steady at a diminished rate then subsequently rises once again.
This trend in insulin secretion represents the exhaustion of the docked insulin stores. The steady,
diminished rate of insulin secretion represents the time necessary to synthesize and secrete more
insulin. In such a case, the signal for insulin secretion outlasts the rate of insulin secretion and
action.



Receptor and transduction pathways of insulin:
Depending on the receptors activated, insulin results in one of two anabolic pathways: 1) the
carbohydrate metabolism pathway or 2) the mitogenic pathway. Insulin mainly targets the liver, adipose
tissue, and skeletal muscle. Insulin stimulates a Growth Factor Receptor, and the proteins that the
receptor activates are called Insulin Receptor Substrates (IRSs), which set off the appropriate signal
transduction pathway.

The Carbohydrate Metabolism Pathway:
In this route, insulin triggers a series of signal transduction pathways that ultimately result in increased
glucose metabolism. First, insulin increases glucose transport into adipocytes and resting muscle cells.
Insulin accomplishes this task by increasing the insertion of GLUT4 transporters into the plasma
membrane of these cells. The family of GLUT transporters are glucose channels that allow for the
passive diffusion of glucose across the cell membrane as dictated by its concentration gradient. The
GLUT4 transporter is inserted in the membrane only in the presence of specific cellular cues. In the
absence of such cues, the GLUT4 transporter resides in a storage pool in the cytosol. In contrast, the
liver cells have GLUT2 transporters that are always present in the plasma membrane; regardless of
insulin stimulation. Thus, glucose transport is independent of insulin in hepatocytes. When blood
glucose levels are high, hepatocytes maintain a concentration gradient for glucose diffusion into the cell
by activating hexokinase, which phosphorylates the glucose converting it to glucose 6-phosphate (G6P).
This allows for steady glucose diffusion into the hepatocyte because G6P is not the same molecule as
glucose. When blood sugar levels are low, the GLUT2 transporters allow for glucose diffusion in the
reverse directioninto the extracellular fluidin order to maintain glucose homeostasis. Noteworthy,
glucose transport in exercising skeletal muscle is also independent of insulin secretion. An unknown
pathway is activated during skeletal muscle contraction that results in increased GLUT4 insertion in the
skeletal muscle plasma membrane. Secondly, insulin enhances cellular utilization and storage of glucose.
This effect is coupled to the first oneInsulin increases glucose uptake in order to drive glucose
metabolic and storage processes. Insulin activates enzymes necessary for glycolysis, as well as those
necessary for glycogenesis and lipogenesis. Insulin also promotes fat synthesis by blocking -oxidation,
which breaks down fats. Thus, insulin increases fat stores in two ways: by activating the enzymes
necessary for fat storage and by inhibiting the enzymes necessary for fat degradation.
The Mitogenic Pathway: Insulin promotes mitosis and therefore increases the number of cell production
and tissue growth. The mitogenic responses work through a series of different proteins involved in a
separate signal transduction pathway.
Metabolic actions of Insulin:
- Increased glycogen synthesis
- Increased glycolysis
- Increase protein synthesis
- Decrease glycogen breakdown by inhibiting glycogen phosphorylase
- Increase lipoprotein lipase
- Increase lipogenesis
- Decrease hormone sensitive lipase
- Inhibit gluconeogenesis
- Adenylate kinase: ADP+ADPATP+AMP. When energy levels are high, the energy levels of AMP
levels are low and vice versa. The levels of AMP regulates AMP kinase (low AMPactivates AMP
kinase and vice versa). AMPK will stimulate the formation of more mitochondria which leads to
higher formation of ATP. AMPK also stimulates glycolysis, stimulates fatty acid oxidation, inhibits
gluconeogenesis, inhibits fatty acid synthesis, etc. Insulin via AKT inhibits this AMPK which will
lead to the metabolic effects of Insulin. Check figure 17-8 from our textbook.
- Adipocytes secrete: Leptin, Resistin, TNF-a, IL-6 which increase insulin resistance whereas
adiponectin decreases insulin resistance.
- Metformin is a drug used to treat Type-II diabetes. This drug stimulates AMPK and inhibits
gluconeogenesis.
The mitogenic pathway involves the effects of insulin as a growth factor and affects all cells. Thus,
insulin leads to the general growth of all cells.
Inhibition of insulin signaling:
- Remove insulin
- Increase phosphatases
- Down-regulation of the receptors