Endocrine Physiology

lecture 2
Dale Buchanan Hales, PhD
Department of Physiology & Biophysics

Metabolic clearance rate

(MCR)
Defines the quantitative removal of hormone from
plasma
The bulk of hormone is cleared by liver and kidneys
Only a small fraction is removed by target tissue
protein and amine hormones bind to receptors and are
internalized and degraded
Steroid and thyroid hormones are degraded after hormonereceptor complex binds to nuclear chromatin

99% of excreted hormone is degraded or conjugated

by Phase I and Phase II enzyme systems

MCR of some hormones

Hormone

Half-life

Amines

2-3 min

Thyroid hormones: T4
T3

6.7 days
0.75 days

Polypeptides

4-40 min

Proteins

15-170 min

Steroids

4-120 min

Hormone-Receptor
interactions
Definition: a protein that binds a ligand with high
affinity and low capacity. This binding must be
saturuable.
A tissue becomes a target for a hormone by
expressing a specific receptor for it. Hormones
circulate in the blood stream but only cells with
receptors for it are targets for its action.

Agonist vs. Antagonist

Agonists are molecules that bind the receptor and
induce all the post-receptor events that lead to a
biologic effect. In other words, they act like the
"normal" hormone, although perhaps more or less
potently
Antagonists are molecules that bind the receptor
and block binding of the agonist, but fail to trigger
intracellular signaling events

Hormone binding study

Hormone-receptor interactions
Hormone--receptor interaction is defined by an
equilibrium constant called the Kd, or dissociation
constant.
The interaction is reversible and how easily the
hormone is displaced from the receptor is a
quantitation of its affinity.
Hormone receptor interactions are very specific
and the Kd ranges from 10-9 to 10-12 Molar

Analysis of hormone interactions:

Scatchard plots

Spare receptors
In most systems the maximum biological response
is achieved at concentrations of hormone lower
than required to occupy all of the receptors on the
cell.
Examples:
insulin stimulates maximum glucose oxidation in
adipocytes with only 2-3% of receptors bound
LH stimulates maximum testosterone production in
Leydig cells when only 1% of receptors are bound

Spare Receptors
Maximum response with 2-3% receptor occupancy
97% of receptors are spare
Maximum biological response is achieved when all
of the receptors are occupied on an average of <3%
of the time
The greater the proportion of spare receptors, the
more sensitive the target cell to the hormone
Lower concentration of hormone required to
achieve half-maximal response

Binding vs. biological response

Spare receptors

Amplification by
2nd messenger

Hormonal measurements
Bioassay
an assay system (animal, organ, tissue, cell or enzyme
system) is standardized with know amounts of the
hormone, a standard curve constructed, and the activity
of the unknown determined by comparison
example: testosterone stimulates growth of prostate gland
of immature or castrate rat in a dose-dependent manner.
Androgen content of unknown sample can be determined
by comparison with testosterone.
disadvantage: cumbersome and difficult
advantage: measures substance with biological activity,
not just amount

Original bioassay systems

defined the endocrine system
Remove endocrine gland and observe what
happened
Prepare crude extract from gland, inject back into
animal and observe what happened
In isolated organ or cell systems, add extract or
purified hormonal preparations and measure
biological response

Hormonal measurements
Chemical methods
chromatography
spectrophotometery

Radioimmunoassay
Radioactive ligand and unlabeled ligand compete for same antibody.
Competition is basis for quantitation
saturate binding sites with radioactively labeled hormone (ligand)
in parallel incubate complex with unknown and determine its
concentration by comparison
cold ligand (standard or unknown) competes with labeled ligand
for binding to antibody and displaces it in a dose-dependent way
amount of cold ligand is inversely proportional to amount of
radioactivity
(cold competes with hot so the more cold that binds antibody the
more hot is displaced resulting in fewer counts being associated
with complex.

radioactivit
y

RIA

Increasing amount of insulin

RIA
advantages:
extremely sensitive due to use of radioisotope
large numbers of samples can be processed simultaneously
small changes in hormone concentrations can be reproducibly
quantitated
Easily automated for high-throughput analysis

disadvantage:
can't determine if hormone measured has biological activity
peptide hormones can be denatured and not active but still retain
their antigenic character

Classes of hormones
The hormones fall into two general classes
based on their solubility in water.
The water soluble hormones are the
catecholamines (epinephrine and
norepinephrine) and peptide/protein hormones.
The lipid soluble hormones include thyroid
hormone, steroid hormones and Vitamin D3

Types of receptors
Receptors for the water soluble hormones are found
on the surface of the target cell, on the plasma
membrane.
These types of receptors are coupled to various second
messenger systems which mediate the action of the
hormone in the target cell.

Receptors for the lipid soluble hormones reside in

the nucleus (and sometimes the cytoplasm) of the
target cell.
Because these hormones can diffuse through the lipid
bilayer of the plasma membrane, their receptors are
located on the interior of the target cell

Hormones and their receptors

Hormone

Class of
hormone

Location

Amine
(epinephrine)

Water-soluble

Cell surface

Amine (thyroid
hormone)

Lipid soluble

Intracellular

Peptide/protein

Water soluble

Cell surface

Steroids and
Vitamin D

Lipid Soluble

Intracellular

Second messenger systems

Receptors for the water soluble hormones
are found on the surface of the target cell,
on the plasma membrane. These types of
receptors are coupled to various second
messenger systems which mediate the
action of the hormone in the target cell

Second messengers for cellsurface receptors

Second messenger systems include:
Adenylate cyclase which catalyzes the conversion of
ATP to cyclic AMP;
Guanylate cyclase which catalyzes the conversion of
GMP to cyclic GMP (cyclic AMP and cyclic GMP are
known collectively as cyclic nucleotides);
Calcium and calmodulin; phospholipase C which
catalyzes phosphoinositide turnover producing inositol
phosphates and diacyl glycerol.

Types of receptors

Second messenger systems

Each of these second messenger systems activates
a specific protein kinase enzyme.
These include cyclic nucleotide-dependent protein
kinases
Calcium/calmodulin-dependent protein kinase, and
protein kinase C which depends on diacyl glycerol
binding for activation.
Protein kinase C activity is further increased by calcium which
is released by the action of inositol phosphates.

Second messenger systems

The generation of second messengers and
activation of specific protein kinases results in
changes in the activity of the target cell which
characterizes the response that the hormone
evokes.
Changes evoked by the actions of second
messengers are usually rapid

Signal transduction
mechanisms of hormones
Activation of
adenylate
cyclase

Inhibition of
adenylate
cyclase

Increased
phosphoinositide
turnover

Tyrosine kinase
activation

-adrenergic

2-adrenergic

1-adgrenergic

Insulin

LH, FSH, TSH,

hCG

Opioid

Angiotensin II

Growth factors
(PDGF, EGF,
FGF, IGF-1

Glucagon

Muscarinic
cholinergic M2

Muscarinic
cholinergic M3

Growth hormone

Vasopressin V1

Prolactin

Vasopressin- V2
ACTH

Cell surface receptor action

G-protein coupled receptors

Adenylate cyclase, cAMP and PKA

Amplification
via 2nd
messenger

Transmembrane kinase-linked
receptors
Certain receptors have intrinsic kinase activity. These
include receptors for growth factors, insulin etc. Receptors
for growth factors usually have intrinsic tyrosine kinase
activity
Other tyrosine-kinase associated receptor, such as those for
Growth Hormone, Prolactin and the cytokines, do not have
intrinsic kinase activity, but activate soluble, intracellular
kinases such as the Jak kinases.
In addition, a newly described class of receptors have
intrinsic serine/threonine kinase activitythis class
includes receptors for inhibin, activin, TGF, and
Mullerian Inhibitory Factor (MIF).

Protein tyrosine kinase receptors

Receptors for lipid-soluble

hormones reside within the cell
Because these hormones can diffuse through the lipid
bilayer of the plasma membrane, their receptors are located
on the interior of the target cell.
The lipid soluble hormone diffuses into the cell and binds
to the receptor which undergoes a conformational change.
The receptor-hormone complex is then binds to specific
DNA sequences called response elements.
These DNA sequences are in the regulatory regions of
genes.

Receptors for lipid-soluble

hormones reside within the cell
The receptor-hormone complex binds to the regulatory
region of the gene and changes the expression of that gene.
In most cases binding of receptor-hormone complex to the
gene stimulating the transcription of messenger RNA.
The messenger RNA travels to the cytoplasm where it is
translated into protein. The translated proteins that are
produced participate in the response that is evoked by the
hormone in the target cell
Responses evoked by lipid soluble hormones are
usually SLOW, requiring transcription/translation to
evoke physiological responses.

Mechanism of lipid
soluble hormone
action

Receptor control mechanisms

Hormonally induced negative regulation of receptors is
referred to as homologous-desensitization
This homeostatic mechanism protects from toxic effects of
hormone excess.
Heterologous desensitization occurs when exposure of the cell
to one agonist reduces the responsiveness of the cell any other
agonist that acts through a different receptor.
This most commonly occurs through receptors that act through
the adenylyl cyclase system.
Heterologous desensitization results in a broad pattern of
refractoriness with slower onset than homologous
desensitization

I fought the law, but the law

won..

Mechanisms of endocrine
disease
Endocrine disorders result from hormone
deficiency, hormone excess or hormone
resistance
Almost without exception, hormone
deficiency causes disease
One notable exception is calcitonin deficiency

Mechanisms of endocrine
disease
Deficiency usually is due to destructive
process occurring at gland in which
hormone is producedinfection, infarction,
physical compression by tumor growth,
autoimmune attack
Type I Diabetes

Mechanisms of endocrine
disease
Deficiency can also arise from genetic
defects in hormone productiongene
deletion or mutation, failure to cleave
precursor, specific enzymatic defect (steroid
or thyroid hormones)
Congenital Adrenal Hyperplasia

Mechanisms of endocrine
disease
Inactivating mutations of receptors can
cause hormone deficiency

Testicular Feminization Syndrome

Mechanisms of endocrine
disease
Hormone excess usually results in disease
Hormone may be overproduced by gland
that normally secretes it, or by a tissue that
is not an endocrine organ.
Endocrine gland tumors produce hormone
in an unregulated manner.
Cushings Syndrome

Mechanisms of endocrine
disease
Exogenous ingestion
of hormone is the
cause of hormone
excessfor example,
glucocorticoid excess
or anabolic steroid
abuse

Mechanisms of endocrine
disease
Activating mutations of cell surface receptors
cause aberrant stimulation of hormone production
by endocrine gland.
McCune-Albright syndrome usually caused by
mosaicism for a mutation in a gene called GNAS1
(Guanine Nucleotide binding protein, Alpha
Stimulating activity polypeptide 1).
The activating mutations render the GNAS1 gene
functionally constitutive, turning the gene irreversibly
on, so it is constantly active. This occurs in a mosaic
pattern, in some tissues and not others.

Mechanisms of endocrine
disease
Malignant transformation of non-endocrine
tissue causes dedifferentiation and ectopic
production of hormones
Anti-receptor antibodies stimulate receptor
instead of block it, as in the case of the
common form of hyperthyrodism.
Graves Disease

Mechanisms of endocrine
disease
Alterations in receptor number and function
result in endocrine disorders
Most commonly, an aberrant increase in the
level of a specific hormone will cause a
decrease in available receptors
Type II diabetes

Hypothalamus and Pituitary

Hypothalamus and Pituitary

The hypothalamus-pituitary unit is the most
dominant portion of the entire endocrine
system.
The output of the hypothalamus-pituitary
unit regulates the function of the thyroid,
adrenal and reproductive glands and also
controls somatic growth, lactation, milk
secretion and water metabolism.

Hypothalamus and pituitary gland

Hypothalamus and pituitary gland

Hypothalamus and Pituitary

Pituitary function depends on the hypothalamus
and the anatomical organization of the
hypothalamus-pituitary
unit
reflects
this
relationship.
The pituitary gland lies in a pocket of bone at the
base of the brain, just below the hypothalamus to
which it is connected by a stalk containing nerve
fibers and blood vessels.
The pituitary is
composed to two lobes-- anterior and posterior

Posterior Pituitary:
neurohypophysis
Posterior pituitary: an outgrowth of the
hypothalamus composed of neural tissue.
Hypothalamic neurons pass through the
neural stalk and end in the posterior
pituitary.
The upper portion of the neural stalk
extends into the hypothalamus and is called
the median eminence.

Hypothalamus and
posterior pituitary
Midsagital view
illustrates that
magnocellular
neurons
paraventricular and
supraoptic nuclei
secrete oxytocin and
vasopressin directly
into capillaries in the
posterior lobe

Anterior pituitary:
adenohypophysis
Anterior pituitary: connected to the hypothalamus
by the superior hypophyseal artery.
The antererior pituitary is an amalgam of hormone
producing glandular cells.
The anterior pituitary produces six peptide
hormones: prolactin, growth hormone (GH),
thyroid stimulating hormone (TSH),
adrenocorticotropic hormone (ACTH), folliclestimulating hormone (FSH), and luteinizing
hormone (LH).

Hypothalamus and
anterior pituitary
Midsagital view
illustrates
parvicellular
neurosecretory
cells secrete
releasing factors
into capillaries of
the pituitary portal
system at the
median eminence
which are then
transported to the
anterior pituitary
gland to regulate

Anatomical and functional

organization

neocortex
Reituclar
activating
substance
Sleep/
wake

Thalamus

Limbic
system

pain

Emotion, fright,
rage, smell

Heat regulation
(temperature)

Water balance (blood

volume, intake--thirst,
outputurine volume)

Energy
regulation
(hunger,
BMI)

Optical
system
vision

Autonomic
regulation
(blood pressure
etc)

Regulation
of
Hypothalamus

Metabolic rate, stress

response, growth,
reproduction, lactation)

posterior
pituitary
hormones

Anterior
pituitary
hormones

Hypothalamus/Pituitary
Axis

Hypothalamic releasing factors for

anterior pituitary hormones
Travel to adenohypophysis via hypophyseal-portal
circulation
Travel to specific cells in anterior pituitary to
stimulate synthesis and secretion of trophic
hormones

Hypothalamic releasing hormones

Hypothalamic releasing hormone

Effect on pituitary

Corticotropin releasing hormone

(CRH)
Thyrotropin releasing hormone
(TRH)
Growth hormone releasing
hormone (GHRH)
Somatostatin

Stimulates ACTH secretion

Gonadotropin releasing hormone

(GnRH) a.k.a LHRH
Prolactin releasing hormone (PRH)
Prolactin inhibiting hormone
(dopamine)

Stimulates TSH and Prolactin

secretion
Stimulates GH secretion
Inhibits GH (and other hormone)
secretion
Stimulates LH and FSH
secretion
Stimulates PRL secretion
Inhibits PRL secretion

Characteristics of hypothalamic
releasing hormones

Secretion in pulses
Act on specific membrane receptors
Transduce signals via second messengers
Stimulate release of stored pituitary hormones
Stimulate synthesis of pituitary hormones
Stimulates hyperplasia and hypertophy of target
cells
Regulates its own receptor

Hypothalamus
and anterior
pituitary

Anterior pituitary
Anterior pituitary: connected to the hypothalamus
by hypothalmoanterior pituitary portal vessels.
The anterior pituitary produces six peptide
hormones:

prolactin, growth hormone (GH),

thyroid stimulating hormone (TSH),
adrenocorticotropic hormone (ACTH),
follicle-stimulating hormone (FSH),
luteinizing hormone (LH).

Anterior pituitary cells and hormones

Cell type

Pituitary
Product
population

Target

Corticotroph

15-20%

ACTH
Adrenal gland
-lipotropin Adipocytes
Melanocytes

Thyrotroph
Gonadotroph
Somatotroph
Lactotroph

3-5%
10-15%
40-50%
10-15%

TSH
LH, FSH
GH
PRL

Thyroid gland
Gonads
All tissues, liver
Breasts
gonads

Anterior pituitary hormones