II. Examining endocrine disruptors
1. Actions of hormones in the human bodyThis chapter is designed to make understanding of subsequent chapters easier by
providing essential information. It describes the physiological roles of endocrineorgans and hormones and the possible mechanisms, that is how the inherent
actions of hormones could be disrupted by exogenous chemicals.
(1)

What is “endocrine”
endocrine”?

Among the various cell groups within the body, many cells create proteins,polypeptides, amines and lipids, and retain them in the cytoplasm in the form ofsecretory granules. Secretion generally refers to extracellular excretion of the
products in these secretory granules. The secretion can be divided into exocrineand endocrine according to the secretory pathway. For example, exocrine cells of
the pancreas produce substances that become digestive enzymes and contents ofdigestive juices, and secrete them into the duodenum through the pancreatic duct.
This process is considered to be a typical external secretion or exocrine.
On the other hand, the pancreas has a cell group called Islet of Langerhans.
Although these cells have secretory granules, they do not have an excretory duct toguide the secreted materials to the place in which they play a role. These secretedmaterials are transported by the circulation to the target organs, where theystimulate and switch on the functions. As they are secreted into the circulation
and are not connected via an external conduit, this mode of secretion is calledinternal secretion or endocrine. In contrast to substances that are carried far totarget organs, substances that act on cells neighboring them can be classified asparacrine and those acting on the secretory cells themselves can be classified as
autocrine.
Substances which are secreted from endocrine glands and act on target organs,tissues, and cells are classically called "hormones". The word "hormone" wascoined by W.M. Bayliss and E.H Starling in 1902 and comes from the Greek "toawaken". Currently it is used to include substances capable of transferring
information. However, among information transferring substances, those secreted5

by cells of the immune system are specifically called "cytokines". The following are
the classical endocrine organs of humans:
1. Thyroid gland
Thyroid gland located in the frontal portion of the neck, under the cricoid
cartilage. This is a butterfly-shaped organ that can be palpated from outside. It
produces thyroid hormones, which play an important role in the growth anddevelopment, in particular, in mental and physical development during theprenatal and infant periods. Its absence brings about severe mental and growth
retardation called "cretinism". In adults, it plays an important role inmetabolism and heat production. An excess of thyroid hormone results intachycardia, tremor, heat intolerance and weight reduction which are typicalsigns and symptoms for "hyperthyroidism".
2. Parathyroid glandThis organ, located next to the thyroid glands, consists of four rice grain-sized
tissues. It secretes parathyroid hormone that mobilize the calcium from thebone to maintain normal calcium concentrations in the blood. Its absence
causes cramp and tetany and is coined as "hypoparathyroidism". An excess ofparathyroid hormone causes an elevation of serum calcium, urinary tract stone,
or decaleificafion of bone and is coined as "hyperparathyroidism".
3. Adrenal gland
Located on the upper part of the kidney, it produces steroid hormones, among
which are glucocorticoids, mineral corticoids and sex steroids, as well as theirprecursors.
1) Glucocorticoids are essential and permissive for vital metabolism and areparticularly necessary for the body to cope with stress such as shock andinfection; the lack of adrenocortical activity causes Addison's disease.
2) Mineral corticoids are necessary to control the body electrolytes and fluidbalance as well as to maintain blood pressure.
3) Sex steroids have been shown to be basic not only to sexual development andbehavior, but also to the maintenance of various tissues such as bone.
4. Ovary
The ovary consists of ovarian follicles, luteal body and connective tissues.

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Ovarian follicles contain ovarian cells and granulosa cells surrounding them.
During ovarian follicle development, the granulosa cells synthesize estrogen.
By the effect of estrogen, one of the follicles grows to the mature ovum andreleases an ovum into Fallopian duct connecting to uterus. After ovulation, theluteal body is formed and progesterone is secreted. The ovarian follicles also
secrete inhibin which suppresses pituitary FSH secretion. If a woman has
normal menstrual cycle, it is warranted that the hormone secretion of the ovary
is normally controlled by dominating pituitary and hypothalamus and ovarial
hormones normally act on the endometrium.
5. Testicle
The testicle consists of seminiferous tubules and interstitial tissues. The
seminiferous tubules have Sertoli cells, and various spermatogenic cells ofdifferent stages ranging from spermatocytes to sperm. Sertoli cells also secrete
inhibin which suppresses FSH. Interstitial Leydig cells secrete testosterone.
6. Pituitary glandThis bean-sized organ protrudes from the base of the brain. It regulates andintegrates the above mentioned hormone secretion. From the hypothalamus,
which is part of the brain, it is connected with the nervous system and thespecial vascular system (pituitary portal system). The hypothalamus functions
as commanding center where neuronal information from the brain andendocrine information from each part of the body are integrated; some of the
newly integrated information is transmitted to the pituitary glands. In
response, the pituitary gland secretes hormones that stimulate the thyroid
gland (thyroid stimulating hormone: TSH), adrenal cortex (adenocorticotropic
hormone: ACTH) and gonads (luteinizing hormone: LH, and follicle stimulating
hormone: FSH) in needs andautomatically control the amount of hormones

secreted from each endocrine organ. The pituitary gland also secretes growth
hormone to secure normal height development; lack of this hormone causes
short stature, while their excess by the neoplasm causes acromegaly. Pituitary
gland also secretes prolactine which stimulates lactation. Excess production ofthis hormone by neoplasma is relatively frequent and inhibit the ovullatory
surge of LH causing amenorrhea in women and also inhibit LH-responsivenessof Leydig cells resulting in testosterone deficiency in man. The posterior lobe of
pituitary secretes antidiuretic hormone (ADH) that reabsorbs water from the7

kidney. Lack of ADH causes diabetes insipidus, in which as much as 10 liters ofurine is produced a day.
7. Hypothalamus
Pituitary gland function is also controlled by a higher regulatory center,namely, hypothalamus. The hypothalamus secretes growth hormone-releasing
hormones (GRH) that simulate growth hormone secretion, thyrotropinreleasing hormones (TRH) that stimulate thyrotropin secretion, corticotropin
releasing hormone (CRH), and gonadotropin-releasing hormones (GnRH) that
stimulate LH and FSH secretion from the pituitary gland. GnRH is secreted ata pulse of once every two hours. This pulse frequency in itself is important for
pituitary LH secretion and the continuous secretion is known to suppress LH
secretion.
In this way, the thyroid, adrenal and gonad hormones are dominated by thepituitary glands, which in turn are dominated by the hypothalamus. On theother hand, the hormones secreted by these endocrine organs act on thepituitary gland and on the hypothalamus in such a way that thyroid hormones
suppress TSH secretion, glucocorticoids suppress ACTH secretion, andestrogen suppresses LH in women, and testosterone suppresses LH in men,
adequately controlling the secretion of hormones from the endocrine organs. In
addition, from the ovary follicles and testicular Sertoli cells, a hormone called
inhibin is secreted and acts to suppress the secretion of FSH. Follistatine, aprotein that binds with inhibin, is present in the blood and acts to control theactions of inhibin. Through this control network, adult human is able to control
hormone concentrations within a limited range, even if a certain amount ofexogenous substance mimicking or inhibiting the hormonal action enters thebody.
8. Others
Besides these classically known endocrine organs, adipose tissues are known tosecrete a hormonal substance, leptin, to control appetite and energy
metabolism; and the heart and vascular system secrete atrial natriuretic
polypeptides (hANP) and endotherine, while the central nervous systemsecretes many peptide hormones, including neuropeptide Y (NPY), CCK and
TRF, etc. Steroid hormones such as dehydroepiandrosterone sulfate (DEHA-S)

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are also shown to be secreted from nervous system.
(2)

Differentiation and formation of gonads

The differentiation and development of gonads proceed as follows: Both male andfemale primoridal gonads are produced from the area called the genital ridge,
which is formed in the initial stage of fetal development. In women, the Fallopian
tubes and uterus are formed from the Muellerian tubes; in men, under thedominance of androgen, the gonads that become the testicles descend and thesemiferous tubules and external genitalia are induced from the Wolffian duct.
In the formation of external genitalia, a symmetrical portion of skin extends towrap the urinary tract and form the penis under the effect of androgen and theinterruption of this process results in retentio testis and hypospadias. This process
is dominated by the effects of androgen. Besides androgen itself, transcription
factors induced by androgen is required for this process. When the transcription
factors, a functional protein bind with a specific sequence of DNA directly or
indirectly and induce organ development, may be not formed due either to 1) lackof androgen, 2) lack of receptors to receive the actions of androgen, 3)abnormalities in the collaborative proteins (co-activators, co-repressors, etc.) that
control the actions of the hormone receptors complex and 4) other causes, various
grade of androgen unresponsiveness or insensitivity is resulted. In its extreme
from, the external genitalia and the body characteristics are female in thepresence of male sex chromatin (XY) and male gonads (testicles). If the degree ofabnormality is slight, abnormalities such as hypospadias or retentio testis areobserved.
At the onset of puberty, the secretion of gonadotropic hormones is induced and thedevelopment of gonads is promoted; in women, ovulation and menstruation begin,and in men, sperm formation starts. In childhood, the secretion of gonadotropic
hormones is suppressed by complex nervous and endocrine mechanisms. When
this suppression is removed at the onset of puberty, sexual maturity is brought
about.
(3)

Chemistry of hormones

Hormones can be roughly divided into water-soluble and fat-soluble categories.

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Water-soluble hormones are either proteins, polypeptides, amines or amino acids;
hormones secreted from the hypothalamus, pituitary and thyroid glands are either
proteins, polypeptides, amines, or amino acids, while adrenal cortex and gonads
secrete steroids which is fat-soluble hydrocarbons. As hormones consisting ofproteins and polypeptides have high molecular weight, they cannot pass throughthe cell membrane. They exert their actions by binding with receptor proteins on
the cell surface membrane, and initiate a specific signal transduction process. For
example, adrenaline and noradrenaline are low molecular size amines but exert
their actions by binding with the receptors on the cell membrane. Thyroid
hormones are a kind of amino acid, but reach the cell nucleus and bind with thenuclear receptors. The structure of these receptors is also very similar to that ofsteroid hormone receptors. On the other hand, steroid hormones can easily passthrough the cell membrane and reach the nucleus, exerting their actions bybinding with the nuclear receptor. In any event, a hormone does not exert its
action until it binds with the receptor protein peculiar to the hormone.
(4)

Regulation of blood hormone concentration

As a hormone acts in proportion to its concentration in the blood, blood hormonal
concentrations are often kept constant and controlled by negative feedback
mechanism. However, a positive feedback mechanism also works to quickly
achieve a high concentration if required. For example, to cause ovulation,
pituitary LH secretion must be increased to a higher level. In this instance,
positive feedback occurs in which estrogen secreted from the ovarian follicles
increases blood concentration, and the latter, in turn, inducing LH secretion. In
addition, information that the hormones give to the target organs is not only
hormone concentrations; in some cases, information regarding the number ofsecretory pulse within a certain time is given as well. For example, GnRH has apulsatile secretion every two hours. This pulsatile secretion stimulate LH
secretion. In contrast, when GnRH is secreted at a constant rate, LH secretion issuppressed. In some cases, various hormone-binding proteins present in the bloodand tissues regulate the fraction of hormones reached to the target cell.
(5)

Binding of hormones with hormone receptors

Hormones exert their actions by binding with the receptor specific to eachhormone. The actions of hormones after binding with hormone receptor proteins10are to start the existing response already programmed in the cells, not to produce
a new response.
For example, when insulin binds with a receptor, phosphoric acid binds with thereceptor which itself becomes an enzyme that phosphorylates proteins called IRS.
This IRS, in turn, phosphorylates various other functional protein and changing
their shapes and functions, for example, accelerating the shuffling of glucose
transporting protein and enhancing the incorporation of glucose into the cells,such as of the muscles. In addition, insulin promotes cell proliferation. As
exemplified by insulin, the signal transduction mechanism is sometimes due tophosphorylation of intracellular proteins. The mechanism is also known in whichtranscription of a target protein is enhanced as the phosphorylated proteins
migrate into the nucleus and bind with its DNA.
On the other hand, fat-soluble hormones directly pass through the cell membraneand enter the nucleus to bind with the receptor. Some hormones bind with thereceptor in the cytoplasm. The hormone-cytoplasm receptor protein complex
migrates to the nucleus and bind to the specific sequence of DNA of the target gene.
In some cases, the receptor protein is bound with a variety of chaperon protein (forexample, estrogen binds with heat shock protein) in the cytoplasm. When thehormone binds with the receptor, the protective chaperon protein is separated, andhormone-receptor complex migrates into the nucleus. The receptor protein
possesses a domain which bind with hormones and another domain that binds
with a specific DNA sequence (in many receptor proteins, the latter has a digitalstructure containing zinc coined as "Zinc finger"). In many cases, the receptor
associated with the hormone (=hormone receptor complex) binds with DNA inpairs (homodimer), but there are cases in which the combination of a receptor
specific to the hormone and a receptor of a different substance (for example, one
receptor for thyroid hormone and the other for retinoic acid) binds with the specificpart of the gene (heterodimer). Many collaborative proteins, such as co-activatorsand co-repressors, bind with these hormone-receptor complex. The processmodulates the efficiency of transcription of a target gene, thereby promoting andsuppressing synthesis of a target protein, respectively. There are cases in which
several nucleus receptors have been found for one hormone (alpha, beta, isomers).
For example, thyroid hormone receptors have alpha and beta isomers withdifferent tissue distribution and different actions. Estrogen receptors also have11alpha and beta isomers and may also have different actions.
Although many hormone receptors for known hormones have been identified there
are some receptors whose binding hormones (ligand) in the body have not yet been
elucidated; they are called "orphan receptors." Orphan receptors have been foundamong membrane receptors and also among nucleus receptors; this is one of the
fields that most rigorous investigation is concentrated at present; this may lead tothe discovery of new hormones. For example, from research on an unknown ligandof the membrane receptor that binds with G-proteins, many new hormones havebeen discovered, such as Ohlexin, which is a new appetite controlling substance,
and a prolactin-secreting hormone different from TRH, which had been unknown
and which has not yet been properly named. The aryl hydrocarbon (Ah) receptor,
which is present in the nucleus, binds with dioxins; however, it is not yet clear
which endogenous substance in the body binds with this receptor, or what it does.
The investigation of the endogenous ligand of Ah-receptor would give us a clue inclarifying the mechanism of action of dioxin in animal and man.
The following are also examples in which endogenous factors that do not normallyexist in the body affect normal hormone actions through receptors and the signaltransduction mechanism. For example, hyperthyroidism of Graves' disease iscaused when the TSH receptor antibody produced as a result of autoimmune
abnormality. The antibody bound with TSH receptor mimics TSH in effect andexerts its actions. Pertussis toxins and cholera toxins bind to G-proteins, which
normally binds hormone-cell membrane receptor complex and initiate signaltransduction, and thus mimics the effect of hormone. It is also known thattamoxifen (anti-estrogen, breast cancer treatment drug) binds with estrogen
receptor and that pharmaceutical agent RU486, binds with progesterone receptors,
to exert their inhibitory actions.
(6)

Manifestation of hormone actions

The cells in the body each have unique characteristics and functions, and all havea copy of the same DNA in their gene. The characteristics and functions of the cells
differ because each cell translates the necessary parts the DNA design andsynthesizes the necessary proteins and enzymes. A hormone may often promote
protein synthesis de novo, but in these cases, the hormone receptor complex binds

12with a certain sequence of DNA on the gene that codes this protein, whose basic
transcription factors are activated by transcriptional control factors, and thenRNA polymerase activity is enhanced by the above described mechanism. As aresult, transcription from DNA to mRNA is promoted. Amino acids are recruited
based on the mRNA code, synthesizing specific proteins. Since both transcriptional
control factors and basic transcriptional factors are proteins, actions ascribed tohormone may well be elicited by transcriptional factors induced by respective
hormones. For example, in the differentiation process of gonads, a series oftranscriptional factors exert their actions in proper sequence, as exemplified by
the effect of AD4BP. Inhibition or deletion of these transcriptional factors is
known to have a result similar to that seen in lack of the effect of hormones.
When an organ is being formed, the orderly synthesis of new proteins andenzymes in proper timing and sequence is important. To accomplish this, manytranscription-controlling factors manifest without mistaking the timing andsequence. It is probable that if an exogenous substance mimicking hormonal
effects enters into body by escaping various controlling mechanism, which isimmature at development stage, sequence of events that dominate normal organ
developmentmay

becomeimpairedor

disrupted,

resultinginvariousdevelopmental abnormalities.
Summary
Cells synthesize and secrete various substances. For example, exocrine pancreatic
cells synthesize digestive enzymes and secrete them into the duodenum throughthe excretory duct. This is called external secretion, or exocrine. However, the cellgroup called the Islet of Langerhans in the pancreas secretes substances such as
insulin and glucagon synthesized in the cells, but does not have an excretory duct,
rather, it secretes the substances into the circulation through the capillary vessels.
This is called internal excretion, or endocrine. Hormones have been defined as
substances secreted into the blood stream from specific endogenous glands andcarried by circulation to act on target organs and express their specific effects.
This mode of action is known as endocrine. Recently, other modes of action, in
which the target cells are located neighboring the hormone-producing cells(paracrine); in which the target cells are hormone-producing cells themselves(autocrine); and in which the target cells incorporate the hormone precursors, and13

manufacture and secrete hormones (intracrine), have become known. In addition,it has been demonstrated that hormones are also secreted by tissues that are
originally not endocrine glands, such as adipose tissues and blood vessels.
Currently substances produced in cells that are involved in the controlling
function the body are arbitrarily called hormones.
Hormones are secreted from endocrine glands [pituitary glands, thyroid glands,
parathyroid glands, pancreas, adrenal cortex, medulla, gonads (testicles, ovaries),etc.], tissues of nervous system such as the hypothalamus; adipose tissues, heartand blood vessels; some hormones are carried into the blood stream by themselves
or by binding with specific binding proteins, and exert their actions on organs. As
a result of these actions, hormones dominate such biological functions as energy
metabolism, development, growth, sex differentiation and reproduction, as well as
regulation of metabolism required to maintain the bodily homeostasis, and also
influence on the immune and nervous systems.
All hormones exert their actions by binding with receptors, some on cell
membranes, while others migrate to the nucleus and exert their actions by binding
with the receptors within the nucleus. Various endogenous and exogenous
chemicals and antibodies are known to affect various processes, such as binding
hormone with transport proteins in the blood, the binding with membrane
receptors, and the signal transduction process is mainly consisting of a cascade ofphosphorylation of proteins and enzymes. The disruption of this process causes
abnormalities and disease. Other types of hormones are known to migrate into the
nucleus and bind with nucleus receptors (which are in themselves proteins; inmany cases, there are a number of different receptors for one hormone), whichthen bind with a specific sequence of the DNA of the gene and alter thetranscription of the specific proteins through the actions of many transcriptioncontrolling proteins. It is also known that endogenous and exogenous chemicals
affect the binding of hormone with nucleus receptors, the binding of hormonereceptor complex with DNA, and the binding of cooperative proteins.

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