Endocrinology &

Metabolism
Introduction

10/14/08 1
 Introduction

Janice Dodd, Ph.D.

Molecular Endocrinology
Department of Physiology

10/14/08 2
 Agenda

• Role of the endocrine system

• Structure and function of hormones
• How hormones are measured

10/14/08 3
 Section 1

• Endocrine system
– overview of endocrine effects
– endocrine glands, target cells
• Hormones
– synthesis, structure, storage, secretion,
and transport

10/14/08 4
 Overview

• Endocrine system controls

– Development and Growth
– Energy regulation (storage & mobilization)
– Internal homeostasis (fluids, ions)
– Reproduction (sex, pregnancy, lactation)

10/14/08 5
 Endocrine action

• Endocrine system
controls physiology
via chemical signals
from one part of the
body to another
• Target cell
undergoes a
biological response

10/14/08 6
 Vocabulary

• Endocrine - borne via bloodstream

• Paracrine - acting in local environment
• Autocrine - acting on secreting cell

10/14/08 7
 more Vocabulary

• Neurocine - neural
cells that release
chemical signals into
the bloodstream

10/14/08 8
 Major Endocrine Glands

• Hypothalamus • Adrenal medulla

• Anterior pituitary • Adrenal cortex
• Posterior pituitary • Pancreas
• Pineal • Ovaries
• Thyroid • Testes
• Parathyroids • Placenta

10/14/08 9
10/14/08 10
 Functional classifications
• Releasing hormones
– From hypothalamus to act on pituitary
• Stimulating (or tropic) hormones
– From pituitary to act on another endocrine
gland
• Non-tropic hormones
– From an endocrine gland to target cells

10/14/08 11
 Releasing, Stimulating and
Non-tropic Hormones
• Hypothalamus releases
Thyrotropin Releasing
Hormone (TRH)
• Anterior pituitary
releases Thyroid
Stimulating Hormone
(TSH)
• Thyroid releases
Thyroid Hormone (T3
and T4)
– Target cells respond
– Feedback (stay tuned)
10/14/08 12
 Endocrine target cells
• Target cells respond
to hormones via
specific receptors
• All cells respond to
multiple hormones
• Receptors may be at
the cell surface or
intracellular

10/14/08 13
 Chemical Classification
of Hormones
• Amino hormones
-derived from tyrosine
• Peptide and protein hormones
- encoded in genes
• Steroid hormones
– derived from cholesterol
• Eicosanoid hormones
- derived from fatty acids (arachadonic acid)
10/14/08 14
 Amino Hormones
• Derived from the amino acid tyrosine
• Includes the catecholamines epiniephrine and
norepinephrine from adrenal medulla
• Includes thyroid hormones thyroxine (T4) and
trioiodothyronine (T3) from thryoid

10/14/08 15
 Thyroid Hormones

• Thyroid pro-hormone is stored as

thryoglobulin as an extracellular colloid
• T3 and T4 can cross lipid membranes
readily (secretion and uptake)
• T3 and T4 are small, hydrophobic and
circulate bound to Thyroxine-binding
globulin (TBG)

10/14/08 16
 Protein & Polypeptide
Hormones
• Transcribed from genes
Insulin Synthesis
• Translated in rough ER,
processed in Golgi,
stored in secretory
vesicles
• May undergo one or
more post-translational
modifications including
cleavage, glycosylation,
disulfide bridging
10/14/08 17
 Subunit Hormones
• Several tropic hormones
made by the anterior pituitary
are glycoproteins that share
a common alpha subunit
• Human chorionic
gonadotropin (hCG) from the
placenta also has the same
alpha subunit
• Specificity is achieved
through beta subunit

10/14/08 18
 Protein Hormones

• Protein and polypeptide hormones

(including glycoprotein subunit
hormones) are stored in secretory
granules and released on stimulation
• Ca++ dependent event – exocytosis
• Hydrophilic - cannot freely cross
membranes

10/14/08 19
 Steroid Hormones

• Not encoded in genes, but derived from

cholesterol through enzymatic reactions
• Cholesterol is converted to pregnenolone
• Pregnenolone moves between mitochondria
and ER and is precursor to all steroids
• Includes Glucocorticoids, Mineralocorticoids,
Androgens, Estrogens and Progestins

10/14/08 20
 
10/14/08
                                                                                       
21
 Enzymes in Steroid
Biosynthesis
• Side-chain cleavage enzyme; desmolase (CYP11A1)
• 3 beta-hydroxysteroid dehydrogenase (3 beta HSD)
• 17 alpha-hydroxylase/17,20 lyase (CYP17)
• 21-hydroxylase (CYP21A2)
• 11 beta-hydroxylase (CYP11B1)
• Aldosterone synthase (CYP11B2)
• Aromatase (CYP19)

• Mutation or failure of any of these genes can lead endocrine

disease
– Eg. Aromatase (CYP19) block will prevent estrogen synthesis

10/14/08 22
 Steroid Hormones

• Steroids are lipophilic molecules that

freely cross membranes
• Steroids are not stored in endocrine
glands but are released as made
• Steroids travel in the blood associated
with Steroid-binding Globulins (SBGs)

10/14/08 23
 Eicosanoid hormones

• Small lipid molecules

• Most abundant precursor is
arachadonic acid
• Includes prostaglandins,
prostacyclins, leukotrienes,
thromboxanes
• Very short half-lives

10/14/08 24
 Terminology Tyranny
• Hormones are often named after the
first site of extraction or first
demonstrated biological effect
• Names are historical records
- don’t limit your thinking!
• Eg. Prostaglandins
• Eg. Follicle-stimulating hormone (FSH)

10/14/08 25
 Summary of Section 1

• Overview of endocrine effects

• Major endocrine glands
• Chemical classification of hormones
• Functional classification

10/14/08 26
 Section 2

• Endocrine system control

– axes and feedback loops
• Hormone measurement
– methods, normal range, fluctuations

10/14/08 27
 Endocrine action

• Endocrine system
controls physiology
via chemical signals
from one part of the
body to another
• Target cell
undergoes a
biological response

10/14/08 28
10/14/08 29
 Hypothalamic-Pituitary Axes

• Three main axes involving hypothalamus and

pituitary control much of endocrine system
- operate by negative feedback
(short and long loops)
– Hypothalamic-Pituitary-Thyroid axis (HPT)
– Hypothalamic-Pituitary-Adrenal axis (HPA)
– Hypothalamic-Pituitary-Gonadal axis (HPG)

10/14/08 30
 Hypo – Pit-Thyroid Axis

10/14/08 31
 Hypo-Pit-Gonadal Axes

10/14/08 32
 Control of Hormone Secretion

• Basic or simple control

• Negative feedback mechanism
• Positive feedback mechanism
• Inhibitory control
• Metabolic control

10/14/08 33
 Basic or simple control

• Limited hormone
made and/or stored
• Full release on
stimulation
• Response is limited
in magnitude or
duration

10/14/08 34
 Negative Feedback Loop
• Most common
control mechanism
in endocrine system
• Eg. thyroid releases
T3 and T4
• Feedback to
hypothalamus (and
pituitary) limits
releasing signal
10/14/08 35
 Positive Feedback Control
• Less common mechanism
• Used when signal
amplification is needed
• Eg. Oxytocin (made in
hypothalamus and released
by posterior pituitary)
• Parturition and suckling

10/14/08 36
 Inhibitory Control

• Hormones that are • Example:

not released until a - Growth hormone
specific inhibitor is release inhibited by
suppressed somatostatin

10/14/08 37
 Metabolic Control
• Activation of bioactive hormone requires
modification (cleavage, de-iodination,
reduction)
• May occur in target tissue
• Eg. 5-alpha reductase
Conversion of testosterone (T) to
bioactive dihydrotestosterone (DHT)

10/14/08 38
 Example of Metabolic Control
• Normally, external genitalia convert T to DHT
and DHT stimulates male phenotype
• In condition referred to as ambiguous genitalia
• XY genotype, testes, male internal
reproductive tract,Testosterone production
• all normal
• T >X> DHT in target tissues
• External genitalia do not virilize

10/14/08 39
 Endocrine Rhythms

• Hormone levels may

fluctuate in
response to external
stimuli (food, light,
activity)
• Circadian and longer
rhythms also exist

10/14/08 40
 Fluctuations

• Changes in hormone levels occur

normally throughout day (and night)
– Accurate measures may require multiple
sampling over time
– Peripheral blood levels are only one
measure; local concentrations may be
higher and more relevant
10/14/08 41
 Measurement of Hormones

• Most used is radioimmunoassay (RIA)

– Nobel prize for development (Yalow)
– Can also be used for other bio-molecules
such as drugs, PSA, disease antigens
• Hormones present in nanamolar to
picamolar range
– Very sensitive tests required

10/14/08 42
 Radioimmunoassay

• Pure radiolabelled hormone (H*) binds to

antibody
• Unlabelled hormone (H) from bio-fluid
competes for binding and displaces H*
• Sensitive, quantiative

10/14/08 43
                                                                                     

            

10/14/08 44
 INSULIN RIA
INTERPOLATION OF UNKNOWN
INSULIN RIA STANDARD CURVE
cpm INSULIN vs [Insulin]

1250
Standard curve made
with pure hormone 1000

CPM INSULIN BOUND

Bio-sample (urine, 750
blood, etc) read
from standard 500

Hormone 250
concentration
determined 0
0 1 2 3 4 5 6 7 8
INSULIN (ng/ml)
10/14/08 45
 Other Hormone Assays
• Radioreceptor assays (RRA)s use
hormone receptors instead of
antibodies
• Bio-assays use target cell outcomes to
measure biological activity of hormone
sample (or biological fluid)
– Cells in culture may respond by expressing
a known gene product
– New science of detecting “endocrine
10/14/08
disruptors” in environmental materials 46
 Summary of Section 2

• Axes and feedback loops

• Control of hormone secretion
• Measurements of hormones

10/14/08 47
 Section 3

• Hormone action
– Receptors and target cell responses
• Endocrine dysfunction
– Points where things go wrong

10/14/08 48
 Endocrine action

• Endocrine system
controls physiology
via chemical signals
from one part of the
body to another
• Target cell
undergoes a
biological response

10/14/08 49
 Mechanism of Hormone
Action
• Alll hormone action is receptor
mediated
• Hormone receptors act as allosteric
effectors
– hormone binding to specific receptor
results in conformational change in
receptor that conveys a signal to target cell

10/14/08 50
 Cell surface receptors

• Peptide hormones and catecholamines

bind to cell surface receptors
• Receptors have extracellular,
transmembrane and intracellular
domains
• Extracellular domain contains ligand
(hormone) binding site

10/14/08 51
 Cell surface receptors

10/14/08 52
 G protein –linked receptors

10/14/08 53
 Non-G linked receptors

• Cell surface receptors with intracellular

domains with intrinsic enzymatic activity
– Protein kinase (serine or tyrosine kinases)
• Or intracellular domains that link closely
to other enzymes

10/14/08 54
 Non-G protein Receptors

10/14/08 55
 Second messengers

• Binding to cell surface receptors releases

second messenger molecules inside cell
• Second messengers include
– cAMP or cGMP
– phopholipids diacylglycerol and inositol
triphosphate (DAG and IP3)
– calcium

10/14/08 56
 cAMP as second messenger

10/14/08 57
 Phospholipids as Second
Messengers

10/14/08 58
 Biological effects

• Second messengers create phenotypic

changes in target cells
– Alter phosphorylation (activity) of proteins
– Alter permeability of membranes
– Indirectly influence gene expression

10/14/08 59
 Receptor Down-regulation

• After hormone binding receptors may

be internalized (coated pits)
• Leads to reduced responsiveness of
target cell (usually temporary)
• Receptor may be recycled to cell
surface or degraded

10/14/08 60
 Intracellular Receptors
• Steroid and thyroid hormones act via
intracellular receptors
• Hormone-receptor complex interacts
directly with DNA in chromatin fibre at
the promoter of specific genes
• H-R complex acts as a transcription
factor to enhance (or decrease) rate of
transcription
10/14/08 61
 Intracellular Receptors

10/14/08 62
 Steroid Receptor Domains

Steroid receptors interact with the grooves

in DNA double-helix via ‘zinc fingers’
formed as loops in the receptor

10/14/08 63
 Response Elements in Genes

• Steroid hormone
receptors recognize
specific DNA
elements in genes
• Short elements are
steroid specific

10/14/08 64
 Endocrine Dysfunction
• At level of endocrine • At level of target cell
gland – Structure, stability of
– Structure, secretion, receptor
stability, elimination of
– Downregulation reduces
hormone
sensitivity
– Primary problems in
– Post-receptor signal
gland of origin
transduction defects
– Secondary due to signals • Second messengers
from Hypo-Pit
• Gene expression
– Presence of other • HRE
agonists / antagonists

10/14/08 65
 Summary of Section 3

• Receptors
• Target cell responses
• Endocrine dysfunction

10/14/08 66
 Where to Get More
Information

• Look at your notes

• Look at your textbook and any recent
textbook of Endocrinology
• Contact me: jdodd@ms.UManitoba.ca

10/14/08 67