The Endocrine

System

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Endocrine Glands
A group of secretory cells that release their
products, chemical signals called hormones, usually
into the circulation.
Fig.1: Endocrine
secretion.
 Endocrine glands include:
 Pituitary, thyroid, parathyroid, adrenal and pineal glands.

 Hypothalamus, thymus, pancreas, ovaries, testes, kidneys,

stomach, liver, small intestine, skin, heart, adipose tissue,
and placenta also have endocrine function.

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 Chemical Structure and Synthesis of Hormones

Three general classes of hormones exist:

1. Proteins and polypeptides, including hormones secreted by the

anterior and posterior pituitary gland, the pancreas (insulin and
glucagon), the parathyroid gland (parathyroid hormone), and many
others.

2. Steroids secreted by the adrenal cortex (cortisol and aldosterone),

the ovaries (estrogen and progesterone), the testes (testosterone),
and the placenta (estrogen and progesterone).

3. Derivatives of the amino acid tyrosine, secreted by the thyroid

(thyroxine and triiodothyronine) and the adrenal medullae (epinephrine
and norepinephrine).

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Chemical Structure and Synthesis of Hormones

Proteins and polypeptides

 Protein and peptide hormones are synthesized on the rough end of the endoplasmic
reticulum of the different endocrine cells, in the same fashion as most other proteins.

 They are usually synthesized first as larger proteins that are not biologically active
(preprohormones) and are cleaved to form smaller prohormones in the endoplasmic
reticulum. These are then transferred to the Golgi apparatus for packaging into
secretory vesicles.

 In this process, enzymes in the vesicles cleave the prohormones to produce smaller,
biologically active hormones and inactive fragments. Secretion of the hormones (as
well as the inactive fragments) occurs when the secretory vesicles fuse with the cell
membrane and the granular contents are extruded into the interstitial fluid or directly
into the blood stream by exocytosis.

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 Chemical Structure and Synthesis of Hormones

Proteins and polypeptides

In many cases, the stimulus for

exocytosis is an increase in cytosolic
calcium concentration caused by
depolarization of the plasma
membrane. In other instances,
stimulation of an endocrine cell surface
receptor causes increased cyclic
adenosine monophosphate (cAMP) and
subsequently activation of protein
kinases that initiate secretion of the
hormone. The peptide hormones are
water soluble, allowing them to enter
the circulatory system easily, where
they are carried to their target tissues.

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 Chemical Structure and Synthesis of Hormones

Steroid Hormones Are Usually Synthesized from Cholesterol

and Are Not Stored

• Although there is usually very little

hormone storage in steroid-producing
endocrine cells, large stores of
cholesterol esters in cytoplasm vacuoles
can be rapidly mobilized for steroid
synthesis after a stimulus. Much of the
cholesterol in steroid-producing cells
comes from the plasma, but there is also
de novo synthesis of cholesterol in
steroid-producing cells. Because the
steroids are highly lipid soluble, once
they are synthesized, they simply diffuse
across the cell membrane and enter the
interstitial fluid and then the blood.

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 Chemical Structure and Synthesis of Hormones

Amine Hormones Are Derived from Tyrosine

• The two groups of hormones derived from tyrosine, the thyroid and the
adrenal medullary hormones, are formed by the actions of enzymes in
the cytoplasmic compartments of the glandular cells.

• The thyroid hormones are synthesized and stored in the thyroid gland
and incorporated into macromolecules of the
protein thyroglobulin, which is stored in large follicles within the thyroid
gland. Hormone secretion occurs when the amines are split from
thyroglobulin, and the free hormones are then released into the blood
stream.

• After entering the blood, most of the thyroid hormones combine with
plasma proteins, especially thyroxine-binding globulin, which slowly
releases the hormones to the target tissues.

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 Mechanisms of Action of Hormones

• The first step of a hormone’s action is to bind to specific receptors at

the target cell. Receptors for some hormones are located on the target
cell membrane, whereas other hormone receptors are in the cytoplasm
or the nucleus.

• When the hormone combines with its receptor, this usually initiates a
cascade of reactions in the cell, with each stage becoming more
powerfully activated so that even small concentrations of the hormone
can have a large effect.

• Hormonal receptors are large proteins, and each cell that is to be

stimulated usually has some 2000 to 100,000 receptors. Also, each
receptor is usually highly specific for a single hormone; this determines
the type of hormone that will act on a particular tissue. The target
tissues that are affected by a hormone are those that contain its specific
receptors.

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 Mechanisms of Action of Hormones

The locations for the different types of hormone receptors are generally the
following:

1. In or on the surface of the cell membrane. The membrane

receptors are specific mostly for the protein, peptide, and catecholamine
hormones.

2. In the cell cytoplasm. The primary receptors for the different steroid
hormones are found mainly in the cytoplasm.

3. In the cell nucleus. The receptors for the thyroid hormones are found
in the nucleus and are believed to be in direct association with one or more
of the chromosomes.

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 Mechanisms of Action of Hormones

The Number and Sensitivity of Hormone Receptors

• The number of receptors in a target cell usually does not remain

constant from day to day, or even from minute to minute. The receptor
proteins themselves are often inactivated or destroyed during their
function, and at other times they are reactivated, or new ones are
manufactured by the protein-manufacturing mechanism of the cell.

• For instance, increased hormone concentration and increased binding

with its target cell receptors sometimes cause the number of active
receptors to decrease.

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 Mechanisms of Action of Hormones

Ion Channel–Linked Receptors

• Virtually all the neurotransmitter substances, such as acetylcholine and

norepinephrine, combine with receptors in the postsynaptic membrane.
This almost always causes a change in the structure of the receptor,
usually opening or closing a channel for one or more ions. Some of
these ion channel–linked receptors open (or close) channels for sodium
ions, others for potassium ions, others for calcium ions, and so forth.

• The altered movement of these ions through the channels causes the
subsequent effects on the postsynaptic cells. Although a few hormones
may exert some of their actions through activation of ion channel
receptors, most hormones that open or close ions channels do this
indirectly by coupling with G protein–linked or enzyme-linked receptors,
as discussed next.

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 Mechanisms of Action of Hormones

G Protein–Linked Hormone Receptors

• Many hormones activate receptors that indirectly regulate the activity of

target proteins (e.g., enzymes or ion channels) by coupling with groups
of cell membrane proteins called heterotrimeric GTP-binding proteins (G
proteins). Of more than 1000 known G protein–coupled receptors, all
have seven transmembrane segments that loop in and out of the cell
membrane.

• Some parts of the receptor that protrude into the cell cytoplasm are
coupled to G proteins that include three (i.e., trimeric) parts—the α, β,
and γ subunits.

• When the ligand (hormone) binds to the extracellular part of the

receptor, a conformational change occurs in the receptor that activates
the G proteins and induces intracellular signals that either (1) open or
close cell membrane ion channels or (2) change the activity of an
enzyme in the cytoplasm of the cell.

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 G Protein–Linked Hormone Receptors

• In their inactive state, the α, β, and γ subunits of G proteins form a complex that
binds guanosine diphosphate (GDP) on the α subunit. When the receptor is
activated, it undergoes a conformational change that causes the GDP-bound
trimeric G protein to associate with the cytoplasmic part of the receptor and to
exchange GDP for guanosine triphosphate (GTP).Displacement of GDP by GTP
causes the α subunit to dissociate from the trimeric complex and to associate with
other intracellular signaling proteins; these proteins, in turn, alter the activity of ion
channels or intracellular enzymes such as adenylyl cyclase or phospholipase C,
which alters cell function.

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 Mechanisms of Action of Hormones

G Protein–Linked Hormone Receptors

• The signaling event is terminated when the hormone is removed and the
α subunit inactivates itself by converting its bound GTP to GDP; then the
α subunit once again combines with the β and γ subunits to form an
inactive, membrane-bound trimeric G protein.

• Some hormones are coupled to inhibitory G proteins (denoted Gi

proteins), whereas others are coupled to stimulatory G proteins
(denoted Gs proteins). Thus, depending on the coupling of a hormone
receptor to an inhibitory or stimulatory G protein, a hormone can either
increase or decrease the activity of intracellular enzymes.

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 Mechanisms of Action of Hormones

Enzyme-Linked Hormone Receptors

• Some receptors, when activated, function directly as enzymes or are

closely associated with enzymes that they activate. These enzyme-linked
receptors are proteins that pass through the membrane only once, in
contrast to the seven-transmembrane G protein–coupled receptors.

• Enzyme-linked receptors have their hormone-binding site on the outside

of the cell membrane and their catalytic or enzyme-binding site on the
inside. When the hormone binds to the extracellular part of the receptor,
an enzyme immediately inside the cell membrane is activated (or
occasionally inactivated).

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 Mechanisms of Action of Hormones

Enzyme-Linked Hormone Receptors

An enzyme-linked receptor-the leptin

receptor. The receptor exists as a
homodimer (two identical parts), and leptin
binds to the extracellular part of the
receptor, causing phosphorylation and
activation of the intracellular associated
janus kinase 2 (JAK2). This causes
phosphorylation of signal transducer and
activator of transcription (STAT) proteins,
which then activates the transcription of
target genes and the synthesis of proteins.
JAK2 phosphorylation also activates
several other enzyme systems that
mediate some of the more rapid effects of
leptin.

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 Mechanisms of Action of Hormones

Steroid Hormones Increase Protein Synthesis

1. The steroid hormone diffuses across the cell membrane and enters the
cytoplasm of the cell, where it binds with a specific receptor protein.

2. The combined receptor protein–hormone then diffuses into or is

transported into the nucleus.

3. The combination binds at specific points on the DNA strands in the

chromosomes, which activates the transcription process of specific genes
to form mRNA.

4. The mRNA diffuses into the cytoplasm, where it promotes the

translation process at the ribosomes to form new proteins.

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 Mechanisms of Action of Hormones

Thyroid Hormones Increase Gene Transcription in the Cell Nucleus

1. They activate the genetic mechanisms for the formation of many

types of intracellular proteins—probably 100 or more. Many of these
are enzymes that promote enhanced intracellular metabolic activity in
virtually all cells of the body.

2. Once bound to the intranuclear receptors, the thyroid hormones can

continue to express their control functions for days or even weeks.

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 Measurement of Hormone Concentrations in
the Blood

• Most hormones are present in the blood in extremely minute

quantities; some concentrations are as low as one billionth of a
milligram (1 picogram) per milliliter. Therefore, it was difficult to
measure these concentrations by the usual chemical means.

• An extremely sensitive method, however, was developed that

revolutionized the measurement of hormones, their precursors, and
their metabolic end products.

• This method is called radioimmunoassay.

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 Measurement of Hormone Concentrations in
the Blood
Radioimmunoassay
• The method of performing radioimmunoassay is as follows. First, an
antibody that is highly specific for the hormone to be measured is
produced.

• Second, a small quantity of this antibody is (1) mixed with a quantity of

fluid from the animal containing the hormone to be measured and (2)
mixed simultaneously with an appropriate amount of purified standard
hormone that has been tagged with a radioactive isotope.

• Therefore, the natural hormone in the assay fluid and the radioactive
standard hormone compete for the binding sites of the antibody. In the
process of competing, the quantity of each of the two hormones, the
natural and the radioactive, that binds is proportional to its
concentration in the assay fluid.

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 Measurement of Hormone Concentrations in
the Blood
Radioimmunoassay

• Third, after binding has reached equilibrium, the

antibody-hormone complex is separated from the
remainder of the solution, and the quantity of
radioactive hormone bound in this complex is
measured by radioactive counting techniques.

• Fourth, to make the assay highly quantitative, the

radioimmunoassay procedure is also performed
for “standard” solutions of untagged hormone at
several concentration levels.

• Then a “standard curve” is plotted. By comparing

the radioactive counts recorded from the
“unknown” assay procedures with the standard
curve. As little as billionths or even trillionths of a
gram of hormone can often be assayed in this
way.

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Measurement of Hormone Concentrations in
the Blood
Enzyme-Linked Immunosorbent Assay

• Enzyme-linked immunosorbent assays (ELISAs) can be used to

measure almost any protein, including hormones. This test combines
the specificity of antibodies with the sensitivity of simple enzyme
assays. This method is often performed on plastic plates that each
have 96 small wells.

• Each well is coated with an antibody (AB1) that is specific for the
hormone being assayed. Samples or standards are added to each of
the wells, followed by a second antibody (AB2) that is also specific
for the hormone but binds to a different site of the hormone
molecule.

• A third antibody (AB3) that is added recognizes AB2 and is coupled to

an enzyme that converts a suitable substrate to a product that can
be easily detected by colorimetric or fluorescent optical methods.

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 Measurement of Hormone Concentrations in
the Blood
Enzyme-Linked Immunosorbent Assay

• Basic principles of the enzyme-linked

immunosorbent assay (ELISA) for
measuring the concentration of a
hormone (H). AB1 and AB2 are
antibodies that recognize the hormone
at different binding sites, and AB3 is an
antibody that recognizes AB2.

• E is an enzyme linked to AB3 that

catalyzes the formation of a colored
fluorescent product (P) from a substrate
(S). The amount of the product is
measured using optical methods and is
proportional to the amount of hormone
in the well if there are excess
antibodies in the well.

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Measurement of Hormone Concentrations in
the Blood
Advantages of Enzyme-Linked Immunosorbent Assay

• Because each molecule of enzyme catalyzes the formation of many

thousands of product molecules, even small amounts of hormone
molecules can be detected.

• In contrast to competitive radioimmunoassay methods, ELISA

methods use excess antibodies so that all hormone molecules are
captured in antibody-hormone complexes. Therefore, the amount
of hormone present in the sample or in the standard is
proportional to the amount of product formed.

• The ELISA method has become widely used in clinical laboratories

because (1) it does not employ radioactive isotopes, (2) much of
the assay can be automated using 96-well plates, and (3) it has
proved to be a cost-effective and accurate method for assessing
hormone levels.

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