8/21/12

Introduction to Endocrine
Endocrine System Hormones Targets Actions (Biological Response) Systems and Signals - nervous system and endocrine system functions are coordinating and regulating - these systems are closely linked and theres no way to separate them - nervous system uses electrochemical signals to send commands to the peripheral nervous system organs and receive info from them - endocrine system produces chemical agents that are transported by the bloodstream to target organs] Responsibilities of the Endocrine System - coordination and communication btw and control of activities throughout the entire body - much slower than nervous system but also control long term processes Classical Endocrine Glands - their primary function is to produce hormones but they also may have other secondary functions o pituitary o thyroid o pancreas o adrenals o gonads o pineal o placenta Nonclassical Endocrine Glands - their primary function is generally metabolic but they can secondarily produce hormones o brain o uterus o adipose o skin o heart o GI tract o Liver o Kidneys Morphological Features of Endocrine Glands 1. Ductless: release hormones directly into the bloodstream not duct system 2. Good blood supply: they are richly vascularized and deliver hormones efficiently into circulation 3. Usually of relatively small size Definition of classical hormone

Must meet requirements of: o Being produced by an organ in a small amount o Being released into the bloodstream o Being transported to a distant organ to exert its specific actions Types of hormones tropic target is endocrine glands because they regulate function of other endocrine system organs; thru these hormones the hypothalamus and pituitary can efficiently coordinate endocrine signaling, complex neuroendocrine pathways nontropic- target is not part of the endocrine system, simple neuroendocrine pathways

Chemical Nature of Hormones hormones derived from single amino acids o amines (epinephrine) o thyroid hormones polypeptides and proteins o polypeptides (ADH) o proteins (TSH) steroids o derivatives of cholesterol (testosterone) arachidonic acid derivatives o prostaglandin

Synthesis and Release of Hormones

Patterns of Hormone Secretion - basal secretion of most hormones is not a continuous process but rather has a pulsatile nature - the pulsatile pattern of hormone secretion is characterized by episodes of release that can be as frequent as every 5-10 minutes or on a yearly basis - each episode is followed by a quiescent period during which plasma levels of the hormone falls toward basal levels, another discharge then occurs and the cycle repeats itself often varying in both frequency and amplitude of the pulses circhoral occur about once an hour ultradian occur in cycles longer than and hour but less than 24 hours circadian occur in a cycle of close to 24 hours quotidian (diurnal) episodes of release of every day circatrigintan episodes of release recur every 30 days circannual (seasonal) - episodes of release take place on yearly basis Hormone Transport Circulate freely: amines, peptides, proteins Bind to carrier proteins: steroids, thyroids nonselective transport proteins: albumin, prealbumin have the capacity to transport nonselectively a variety of low molecular weight hormones specific transport proteins: globulins, high affinity binding for the hormones they carry (thyroid binding globulin, cortisol binding globulin, testosterone binding globulin)

Hormone metabolism most hormones are cleared by the kidney and liver

o degradation by a variety of enzymatic mechanisms (hydrolysis, oxidation, hydroxylation, methylation, decarboxylation, sulfation, glucuronidation ) intracellular degradation of hormones by tar get cells small fraction is excreted intact in the urine and feces

Metabolic Clearance Rate volume of plasma cleared of the hormone/unit time radioactive hormone injected until you reach a steady level and then the disappearance rate is determined and plasma half life is calculated (half life is related to the reciprocal of MCR) binding of a hormone to carrier proteins has a profound effect on the hormone MCR; the greater the binding capacity of the specific protein carrier the slower the clearance rate thsu hormones that circulate mostly in the bound form disappear much more slowly than do hormones that circulate freely or weakly bound (thyroid hormone half life is about day but most proteins range from a few minutes to a few hours) Function of Hormones cell membrane receptors: protein hormones and amines nuclear membrane receptors: steroids and thyroids a single hormone can exert various effects in different tissues a single function is regulated by several hormones a complex function is regulated by multiple hormones

Four Physiological Areas 1. Reproduction: growth of reproductive organs, production of gametes, patterns of sexual behavior, phenotypic differences btw the sexes, continuation of the species 2. Growth & Development: timing and progression of growth 3. Maintenance of Internal Environment: extracellular fluid volume and composition, bp, plasma and tissue level of calcium and phosphate ions, maintenance of bone, muscle and body store of fat 4. Regulation of Energy Balance: food intake, energy expenditure, body weight, metabolic rate Paracrine hormones act on contiguous cells Autocrine hormones act on the same cell that produce them Juxtacrine am membrane bound hormone precursor on one cell is cleaved to yield the active form of the hormone which then interacts with and adjacent cell Endocrine Control System 1. Simple Control Hormone is only produced in small amounts for short time and often that target tissue can only respond to certain levels of hormone 2. Negative Feedback

After hormone B reaches a certain level it turns off production of hormone A at the gland 3. Positive Feedback Positive loop carries on until stimulus stops Suckling at a nipple sends a neurosignal to the brain then the posterior pituitary which releases oxytocin and causes milk ejection at breast 4. Inhibitory Control Hormones with broader action perhaps which do not result in the release of products are often controlled by inhibitory hormones, suppression of inhibitory hormones lead to target hormone release 5. Metabolic Control Some hormone are converted to active form by metabolism, may be regulated locally the amount of hormone produced and the frequency of the secreting episode are modulated by : o the basic mode of release (amplitude and frequency) o specific negative and positive feedback mechanisms

Endocrine Diseases 1. Hormone Deficiency - Pathological conditions of endocrine glands (impaired hormone production) - Genetic defects: deletion or inactivating mutations of genes involved in hormone synthesis, release 2. Hormone Excess - Overproduced by endocrine glands (tumor of endocrine glands) - Produced by non-endocrine tissue (cell dedifferentiation) - Activating mutations of genes involved in hormone production 3. Hormone Resistance - The hormone is present in increased or normal amounts but the expected actions of the hormone do not occur - Genetic mutations of hormones (structurally abnormal) - there are antibodies to the hormone or hormone receptor - receptor defects - defects in the post-receptor mechanism of hormone action 8/23/12 Hormone Receptors a molecule that can recognize and bind to a particular hormone with high affinity and specificity, after binding the hormone the receptor is biochemically and structurally altered, the activated receptor mediated all the actions if the hormone on the cell cell membrane receptor hormones that rare water soluble, the activated membrane receptor initiate signal transduction cascade that result in changes in enzyme activities and alterations in gene expression nuclear membrane receptor the steroid and thyroid hormones as well as retinoids and vitamin D3 diffuse thru the lipophilic plasma membrane of the cell and interact with receptors that are primarily within the nucleus on activation receptors alter transcription of genes

Affinity o Hormone receptor interaction is non-covalent binding (hydrophobic and electrostatic)

o The affinity of the receptor for a particular ligand in relation to its affinity for other related molecules determines the specificity of the hormone receptor interaction o The affinity o hormone receptor interaction is defined in terms of Kd Kd indicates the strength of binding btw A (ligand) and B (ligand) in terms of how easy it is to separate the AB complex If high concentration of A & B is required to form AB this indicates strength of binding is low, the Kd therefore would be higher The smaller the Kd the stronger the binding Hormone Concentration, number of receptors and biological response - the biological response of a target cell to a hormone is determined by o concentration of hormone o concentration of plasma o affinity of the hormone receptor interaction - there is a finite number of receptors for a given hormone on a target cell, normally the receptors on a target cell are almost never saturated and in increase in the concentration and number of occupied receptors and response of target to the hormone is directly proportional to the occupied number of receptors if the maximum response has not yet been achieved - in mot cases however the maximum response of a target cell is achieved at a concentration lower than those required to fully occupy all of the receptors on that cell Alterations in Receptor Number - changes in the concentration of cell receptors for a specific hormone can markedly alter the sensitivity of the target cell to that hormone At receptor level - as you increase the concentration of receptors can decrease sensitivity to the hormone whereas an increase in the receptor concentration cam increase sensitivity of the target cell to the hormone At hormone level - the concentration of hormone itself can regulate the concentration of its own receptors on target cells, an increase in levels of hormones will cause decrease in the receptors Homologous down regulation or desensitization - Negative regulation of receptors o clustering of hormone receptor complexes in coated pits on cell surface o internalization within coated vesicles o degradation by lysosomal enzyme Heterogeneous desensitization - incubation with one agonist reduces the responsiveness of a cell to a number of other agonists that act thru different receptors - this is mostly observed with receptors that act thru the adenylyl cyclase system - this reflects a broad pattern of refractoriness (sensitivity to further stimulation that develops as a result of intense or

prolonged stimulation) that has a slower onset than homologous desensitization Hormone-receptor interaction - agonist: hormones are analogs that bind t receptors and elicit the same biological response as naturally occurring hormone - partial agonist: bind to receptor but less biologically active than native hormone - competitive antagonist: bind to receptors but fail to elicit the normal biological response - partial antagonist: bind to receptors but not completely preventing the normal biological response reduced by native hormones G coupled receptors receptors for polypeptide hormones, prostaglandins, and neurotransmitters a large family of related receptors all contain seven hydrophobic regions each of which forms alpha helix that spans seven times receptor molecule interacts within the plasma membrane with a specific member of G protein family, all G proteins are heterotrimers composed of , and subunits. The subunit is unique to each G protein whereas the and subunit are similar

Tyrosine Kinase Receptor Superfamily - receptors for EGF, insulin, and IGFs - the intracellular domain contains protein kinase domain that specifically stimulate phosphorylation of tyrosine residues on target proteins binding to receptor causes conformational changes involving receptor dimerization that leads to tyrosine kinase activation and subsequent stimulation of downstream signaling pathways Cytokine Receptor Superfamily - multiple subunits - no intrinsic activity - signal transduction mediated by tyrosine kinase activity (Jaks) - receptors for GH, PRL, hematopoietic growth factors Hormone Signaling Pathways Cell Membrane Receptor/ Second Messenger Pathways - cAMP Pathway - phosphatidyl inositol - MAP kinase pathway - Jak/STAT pathway Nuclear Receptors - Thyroid hormone and estrogen (steroid receptors)

Adenylyl Cyclase System

the hormone binds to a specific receptor on cell surface and this results in activation of a membrane bound enzyme or effector (adenylyl cyclase) which generates a soluble extracellular second messenger which then transmits info to the cellular machinery resulting in a biological response

cAMP dependent protein kinase

cAMP dependent protein kinase holoenzyme is composed of a regulatory subunit dimer and two catalytic subunits - the R subunits is a cAMP binds protein while the C subunit when free of R expresses protein kinase activity - the binding of cAMP to R causes dissociation of the inactive holoenzyme to yield two active catalytic subunits - the free active catalytic subunits can now catalyze the phosphorylation of cellular proteins Phosphatidyl inositol pathway

Phosphat idy l Inosit ol Pat hways

ho rm one ho rm one

Blood receptor tyrosine kinase receptor Cell mem br ane G GTP p rot ein Cell

PI3 -Kinase

p85 p110

p hosp holipase C

ph osp hat idyl ino sit o l ( PI)

ph osp hat idyl ino sit o l pho sphat es ( e.g. PIP2 )

ino sit o l t r iph osp hat e ( IP3 )

diacylglycerol (DAG)

pro t ein kinase C

AKT

Akt -Kinase

[ Ca2 + ]
arachid onic acid d erivat ive s

S6 Kin ase

calmo dulin

prot ein sy nt hesis

Diversity of G protein coupled receptor

Enzy me Act ivat ion

MAPK Signaling Cascade

MAPK family of Ser/Thr protein kinases are widely conserved among eukaryotes and are involved in many cellular programs such as cellular proliferation, cell differentiation, cell movement and cell death MAPKKKs are activated in interaction with family if small GTPases and other protein kinases, MAPKKs are phosphorylated and activated by MAPKKKs, and MAPKS are phosphorylated and activated by MAPKKs

8/23/12

Nuclear Receptors
Nuclear Receptor Family - primarily located within the nucleus - ligand activated transcription factors - 48 identified in the human genome - highly conserved across species although larger # found in lower organisms receptor steroid hormone receptors ER GR estrogen receptor glucocorticoid receptor examples of ligands estradiol cortisol

MR AR PR thyroid hormone receptors retinoid receptors vitamin D receptor TR RAR RXR VDR LXR lipid sensors FXR

mineralocorticoid receptor androgen receptor progesterone receptor thyroid hormone receptor retinoic acid receptor retinoid X receptor vitamin D receptor liver X receptor farnesoid X receptor peroxisome proliferator activated receptor

aldosterone testosterone progesterone triiodothyronine (T3) all-trans-retinoic acid 9-cis-retinoic acid 1,25-dihydroxy-vitamin D3 oxysterols (e.g. hydroxycholesterols) bile acids (e.g. chenodeoxycholate) fatty acids, eicosanoids (LTB4, PGJ2*)

PPAR

PPAR

Cellular Localization of Nuclear Receptors in the absence of ligand binding o some nuclear receptors are present within the nucleus and bound to their respective response elements o some are present within the cytoplasm and bound to a complex of heat sock protein on ligand binding o they are all within the nucleus bound to hormone response elements Nuclear Receptor protein Structure/ Function N term: amino terminus of receptor protein A/B: ligand independent transactivation factor C: DNA binding domain containing two zinc finger complexes D: hinge region, less conserved, flexibility E: ligand binding, dimerization, transactivation function, cofactor interaction F: less conserved region C term: carboxyl terminus DNA Binding Domain Contains two repeated unite each folded into a finger like structure with 4 cysteines that coordinate one zinc ion Loop is 11-15 amino acids Spacer region of 15-17 amino acids btw two fingers Binding fingers have the capacity to insert into half turn of DNA D box is responsible for dimerization and recognition of the spacer sequence P box is responsible for recognition of HREs

Hormone Response Elements Specific regions of DNA where receptors bind Type 1 Receptors o Bind as homodimers o Most steroid hormone receptors: GR, MR, AR, PR o Bind to an identical core HRE which consists of a double stranded inverted repeat or palindromic sequence of six nucleotide separated by spacer of 1-3 nucleotide Type 2 Receptors o Bind as heterodimers with retinoid X receptors (RXR) o TR, RXR, RAR,VDR o Bind to a double stranded direct hexameric repeat separated by spacer of 1-5 nucleotides Retinoid X Receptors 3 isoforms form homodimers and work alone a well ligand is the 9 cis retinoic acid, found in the body Orphan nuclear receptor their ligands are unknown Histone Acetylation histone tails not acetylated: chromatin closed, gene off histone tails acetylated: chromatin open, gene on

NR Cofactors: co-activators and co-repressors NR co-activators upon binding their ligands nuclear receptors such as steroid receptors undergo a conformational change of the C terminal transactivation domain AF2 they then bind to their target DNA and recruit co-activators and general transcription factors thru AF2 How do transcriptional co-activators like SRC-1 work SRC: a transcription activator for steroid receptor family, a histone acetyltransferase SCR-3: over expressed in many breast cancers and therefore may be important to tumorigenesis Co-activators may also interact with and recruit other transcriptional co-activators such a P300 and CBP, sometimes known a co-integrators NR co-repressors Distinct family of proteins that may be involved in silencing nuclear receptors in the absence of ligands Include N-coR and SMRT, they usually dissociate from the receptor when the ligand binds, the repress transcription by recruiting histone deacetylases Regulation of gene expression by homodimer receptors After binding hormone the receptor dissociates from a complex with heat shock protein and other proteins Translocates into nucleus and forms a homodimer and binds to its HRE upstream if the target gene

The activated receptor dimer can now recruit a complex of coactivators containing histone acetyltransferase activity, this results an opening of chromatin structure which facilitates assembly of pre-initiation complex, the binding of RNA pol and activation of transcription

Regulation of gene expression by heterodimer receptors - in its silent state the receptor binds to its HRE as a homodimer or as a heterodimer with a molecule of RXR - the T3R interacts with a complex of co-repressors containing histone deacetylase activity which silences transcription by promoting a closed chromatin structure and preventing formation of a stable pre-initiation complex - on binding of T3R the receptor undergoes a conformational change resulting in the dissociation of the co-repressors and association of the co-activators containing acetyltransferases, this in turn facilitates assembly of pre-initiation complex binding of RNA pol II and activation of transcription initiation Activation of gene Transcription by Nuclear Receptor - In silent state o NR recruits co-repressor o Histone is deacetylated o Chromatin is closed off o Gene transcription is off - In activated state o NR recruits co-activators o Histone is acetylated o Chromatin is open o Gene transcription is on Ligand Independent Activation of NRs - Phosphorylation of NR by protein kinases (mainly A/B domain) o CDK o MAPK o PKC o PKA o Akt/PKB Positive or Negative Effects of NR phosphorylation on transcription o PRER, AR, AND PPAR are phosphorylated in the A/B domain by MAPK and this increases transcription o PKA/PKC phosphorylation of the DBD and inhibits dimerization and HRE binding o MAPK action of LBD of RXR inhibits the binding of RAR-RXR and VDR-RXR heterodimer to co-activators

Interaction of NR with cofactors drug development - The anti- estrogen ralifozen changes ER interaction with coactivators Non-Classical effects for NR and or their ligand hormones 1. Classical effects (genomic actions) nuclear receptors act to regulate transcription of genes Actions occur within the nucleus Effects are relatively slow (20 mins) 2. Rapid or nongenomic effects Actions occur outside the nucleus Effects are mediated by known nuclear receptors or other non nuclear receptors Effects occur within second to minutes

An example: ANGELS = activators of nongenomic estrogen like signaling Estren is an ANGEL Acts on bone within 1-2 minutes Activates protein kinases Kinases have rapid non genomic effects Kinases also have slower genomic effects by promoting phosphorylation of TF (including ERs)

Why is it important? Hormone replacement therapy uses sex steroids including estrogen o Side effects Selective estrogen response modulators synthetic drugs o Less potent than HRTs o Act thru classical pathway Alternative therapies for future o ANGELs such as estren may work on bone without affecting reproductive system 8/28/12

Techniques in Endocrine Research and Diagnosis

1). Plasma Hormone Concentration Assays Used to diagnose abnormalities in hormone production Methods: o Immunoassays Competitive Immunoassay Sandwich Immunoassay o Chromatography High performance liquid chromatography o Spectrometry Mass Spectrometry Competitive Assay Competitive inhibition of a labeled antigen to the antibody by unlabeled antigen The unknown concentration of hormone is estimated by allowing it to compete with a known amount of labeled hormone for binding Of the amount if the antibody and labeled hormone is fixed the amount of labeled hormone to antibody depends on concentration of unlabeled hormone

RadioImmunoAssay Hormone is labeled with a radioactive isotope Traditional method of measuring plasma concentration and commonly used in clinical and research labs Standard curve is needed Competition assay Very sensitive Determined concentration of hormone does not always correlate with biological activity Need a good specific antibody Need a set of standards Need to separate the labeled hormone antibody complex from the labeled free hormone after equilibrium

Sandwich Immunoassay/Immunometric Assay

Ab1 is the capture antibody and is bound to a solid phase. It recognize a specific epitope on the antigen and captures it Ab2 is the labeled antibody that recognizes a different epitope on the antigen and allows a signal to be measured and quantified the signal antibody can be altered to yield different signals and different assays : o immunoradiometric use radioisotope o immunochemiluminometric Ab is chemiluminometric o immunofluorometric molecule bound to Ab is fluorescent o immunoenzymetric molecule bound to Ab is capable of enzymatic activity does not rely on the competition of unlabeled hormone with a labeled hormone for a limited amount of antibody the antibodies are used in excess to ensure that the antibody binds all of the hormone in the sample double antibodies more sensitive than competitive immunoassay because uses 2 antibodies faster than competitive because competitive requires time to allow equilibrium to be reached more specific than competitive immunoassay

Enzyme Linked Immunosorbed Assay (ELISA) immobilized capture Ab conjugated to plate surface, antigen is added and add enzyme labeled detection Ab use blood, urine, and saliva as sample a single random sample of hormone whose plasma concentration is relatively stable multiple plasma samples at intervals for hormones secreted in short pulses and pool aliquots for a single determination the result must be interpreted relative to time of samplings of the hormones with a diurnal or sleep related secretion pattern 2). Bioassays Hormone Receptor Binding Assay o Radioreceptor Assay (RRA) A competition assay that is similar to radioimmunoassay except it uses the receptor containing tissue instead of antibody Receptors: Come from target tissue Have high affinity Estimate the biological activity of a hormone and its receptor

Hormone Biological Activity Assay o Nb2 cell proliferation assay If have a hormone that is similar in action to one that already exist, prolactin in this case, you take Nb2 cells that

rare known to express the receptor and see of they have similar activity upon binding

o Frog skin Assay For melanocyte stimulating hormone you measure the color change on the fog skin where it causes a dramatic darkening due to granules of melanin spreading thru the branches if melanophores o Surgical Assay Parabiosis Cross circulation experiment where obsess was joined thru blood stream with normal mouse The obsess mouse had a leptin mutation and without leptin it cant properly regulate fat metabolism but when crossed with normal mouse it received sufficient amounts of leptin to regulate fat metabolism then over time the mouse was no longer obese Western Blot Denatured gel electrophoresis, blot on membrane incubate with primary antibody wash then incubate with enzyme linked secondary antibody, wash and add substrate Measure hormone protein expression and tell if the hormone is expressed in a particular tissue By autograph intensity can estimate how much there is Immunohistochemistry Bind primary specific antibody then bind fluorescent secondary antibody then examine under microscope to detect tissues, cells or subcellular components that express protein Determine if tissues expresses the hormone receptor and distribution of hormone receptor and which cells express it Audioradiography Inject animal with radioactive hormone, wait to examine histology, cover sections with photographic emulsion and expose to film, observe in cell nucleus 8/30/12

Pituitary Gland (1)

Involved in: Growth, metabolism, reproduction, stress response, lactation Hypothalamus Part of the brain located under the thalamus (processing center) wrapped around the third ventricle

Almost every part of the CNS communicates with it Sole link between the nervous and endocrine systems Secretes hormones that control the pituitary gland, located directly below it, and together they control may hormones Integrates activities of nervous and endocrine systems via pituitary Produces releasing hormones that turn on and inhibiting hormones that turn off hormone production at pituitary Hypothalamus hormones are named for the pituitary hormone they regulates Hypothalamus hormones are mostly small peptides and a dopamine Hypothalamus nuclei o Hypothalamus contains several nuclei and nerve tracts o The nuclei that connect with the posterior pituitary are supraoptic nucleus (SON) and paraventricular nucleus (PVN) o They hypothalamus can secrete hormone and store it in the posterior pituitary til its released

Pituitary Gland Located directly under the hypothalamus and connected to it by the pituitary stalk Small gland that varies by species Two developmentally and functionally different parts: o Adenohypophysis Anterior pituitary: contains pars tuberalis, par distalis, pars intermedia o Neurohypophysis Posterior pituitary: contains median eminence, infundibular stalk, and pars nervosa 2 developmental origins

Anterior Pituitary Derived during embryological development from the roof of the mouth Connected to the hypothalamus via portal system (Hypothalamic Pituitary Portal System) No direct nerve supply Most of pituitary hormones are releases from anterior pituitary Posterior Pituitary Derived from nerve tissue The nervous connection to the hypothalamus 2 protein hormones released: oxytocin and vasopressin

2 Important Points 1. hormones released from posterior pituitary are synthesized in the hypothalamus 2. hormones synthesized in the anterior pituitary are dormant unless directed to be released by the hypothalamus via releasing hormones

Hypothalamic Pituitary Axis

integrates activities of nervous and endocrine systems core of the neuroendocrine system

Posterior Pituitary an extension of the hypothalamus composed of glial elements, unmyelinated nerve fibers and axon terminals of neurons whose cell bodies reside in the SON and PVN responsible for secreting two peptides both of which are synthesized in the hypothalamus Posterior Pituitary Hormones both AVP and OT are nonapeptides both circulate in the blood unbound to other proteins both are rapidly removed from circulation mainly by kidney, liver and brain made by neurons, sent to posterior pituitary by pituitary stalk thru axonal transport and stored Oxytocin targets mammary gland to stimulate milk ejection

targets smooth muscle of uterus during labor and enhancing contraction aiding in childbirth may establish maternal behavior also present in males, present at ejaculation physically stimulated can be inhibited by stress made as a precursor and processed as its passed to the posterior pituitary

Mechanism of action of OT recognized by G protein coupled receptor in target tissue receptor activates the IP3/Ca2+ pathway calcium increases, then kinases activated, thru myosin/actin contraction stimulated ADH target the kidneys and induce them to retain water (reducing urine volume) in excess can cause restriction of arterioles increasing arterial pressure regulated by plasma osmolality stimulated by hypothalamus osmoreceptors deficiency causes hypothalamic diabetes insipidus, nephrogenic diabetes occurs when kidneys fail to respond to ADH, both indicated by over production of urine made as a precursor and processed as its passed to the posterior pituitary main role is antidiuresis but has other roles: o promotes ACTH secretion o promotes liver conversion of glycogen to glycogen phosphate o facilitate memory consolidation and retrieval o vasoconstriction mechanism of action of ADH in renal tubules AVP receptor affects soluble intake in the ascending loop of Henle and water reabsorption in the collecting duct, AVP makes distal convoluted tubules and collecting duct permeable to water increasing reabsorption and decreasing volume of urine AVP receptors expressed in distal convoluted tubule, collecting duct

AVP Receptors 1. VR1 G protein coupled

Activates IP3/Ca2+ pathway Many subtypes Expressed in all tissues except the kidney Vasoconstriction

2. VR2 G protein coupled Activate cAMP pathway Kidney receptors Leads to aquaporin insertion in membrane increasing permeability decreasing urine flow VR3 G protein coupled Activate cAMP pathway Expressed in anterior pituitary gland Stimulation of adrenocorticotropic release

Posterior Pituitary associated diseases Diabetes Insipidus AVP Deficiency (central D.I. ) o Genetic AVP deficiency (mutation of AVP gene) o Acquired AVP deficiency (destruction or dysfunction o f hypothalamus-pituitary complex) o Polyuria o no concentrated urine produced o polydipsia o Treatment AVP analog (desmopressin) Specific V2 receptor agonist Longer half life More potent than natural AVP Kidney Hyporesponsiveness to AVP (peripheral AVP) o Genetic hyporesponsivenes (mutation of AVO receptor gene or aquaporin gene) o Acquired hyporesponsiveness (renal injury, drug side effects) o Polyuria o no concentrated urine produced o polydipsia o Treatment:

Antidiuretic drug which increases sensitivity in kidney tubules to AVP

Syndrome of Inappropriate ADH Secretion (SIADH) ADH excess Can be caused by anesthetics, drugs, some tumors secrete A like substances Leads to water retention o Mild no symptoms o Severe coma, convulsion, death Treatment o Control water intake o Decrease or control AVP release o Block AVP action on the kidneys Treatment of Endocrine Diseases Defective or missing hormone Small molecule (thyroid, AVP, estrogen) easier to make and take oral delivery Large molecule (insulin) harder to make and replace, injection delivery Defective or missing receptor Gene therapy Use molecule that act downstream 9/ 4/12

Pituitary (2)
Distinct Cell Populations in Anterior Pituitary

Growth Hormone (GH) Secreted by somatotropes, 191 AA protein, regulates linear growth Prolactin (PRL) Secreted by lactotropes, 198 AA protein, regulates lactation Adrenocorticotrophic hormone (ACTH) Secreted by corticotropes, 39 AA polypeptides, regulates adrenal function Thyroid Stimulating Hormone (TSH) Secreted by thyrotropes, 207 AA, glycoprotein dimer, regulates thyroid gland function

Luteinizing Hormone (LH) Secreted by gonadotropes, 204 AA, regulates corpus luteum formation, estrogen/progesterone, secretion, regulates androgen secretion Follicle Stimulating Hormone (FSH) Secreted by gonadotropes, 204 AA, glycoprotein dimer, regulates growth of ovarian follicles, regulates spermatogenesis

Growth Hormone/Prolactin Superfamily Humans Multiple CS and GH genes on human chromosome 17 o GH expressed in the anterior pituitary o GH-variant expressed in the placenta o CS-A - expressed in the placenta o CS-B - expressed in the placenta o CS-L - expressed in the placenta Mouse Three Classical Members o Prolactin o Placental Lactogen I o Placental Lactogen II Nonclassical members including prolactin like proteins and prolactin o dont interact with the prolactin receptor o most are expressed in the placenta the biological significance of having so many hormones is because you have a very efficient reproductive system

Growth Hormone regulates growth and metabolism hypophysectomized (pituitary removed) rats fail to grow o pituitary extracts reversed this o some growth hormone in the pituitary made by somatotrope cells of the anterior pituitary circulates in the plasma bound to GH binding protein a cleavage product of the extracellular domain of the GH receptor

the somatropes are the most abundant cell in the pituitary gland, they store GH in amounts representing 4-8% of the gland by weight secreted in a pulsatile fashion throughout life the secretion pattern is ultradian rhythm with a frequency of about one pulse every 2-3 hours episodic secretion is influenced by age, sex and sleep o sex women secrete more GH than men and have a higher baseline and larger bursts o age birth elevates during the first few days puberty increases paralleling changes in height velocity and sexual maturity adulthood returns to levels before puberty and stays stable throughout at least the decade of life aging decreases o sleep the most prominent episodes of GH release occur at night during the onset of deep sleep

Central Control Of GH

GRF or GHRH 44 AA peptide neuropeptide released by the hypothalamus in the brain recognizes a 7 transmembrane helix receptor (GRFR) promotes somatotrope cell proliferation and GH secretion GHRH Receptor G protein coupled receptor Expressed only in the anterior pituitary Activates a cAMP second messenger system in somatotropes SST (Somatostatin) 14 AA released from hypothalamus inhibits GH secretion from somatotropes SST receptor inhibits cAMP system in somatotropes Also plays role in gut, pancreas, etc. Synthetic GH secretagogues One small synthetic molecule that acts as GH secretagogues Act like GRF to promote GH secretion May activate IP3/Ca2+ pathway

Potently activates GHS-R

Neurotransmitter - catecholamines o adrenergic- stimulate GH secretion o adrenergic inhibits GH secretion o work by modulating GHRH and SST output Peripheral Control of GH Secretion Ghrelin 28 AA peptide made mostly in the endocrine neurons of the stomach, also in hypothalamus cause GH release (but not PRL, TSH, ACTH, LH, or FSH) so its specific can specifically activate GH secretetagogues receptor (GHSR) and cause subsequent cAMP increase and Ca+ release the GHS-R is a G protein coupled receptor and is found in the pituitary gland and the hypothalamus Glucocorticoids exert a dual effect on GH secretion, first they stimulate and then they inhibit Thyroid Hormone Gonadal Steroids Leptin Indirect Control of GH Secretion Metabolic Signals glucose inhibits GH secretion, hypoglycemia stimulates GH secretion o insulin administered to produce hypoglycemia results in a prompt increase in GH secretion so because of this insulin induced hypoglycemia is used a s a proactive test to stimulate GH secretion in individuals with suspected GH deficiency o mediated in part by Ghrelin certain AAs given orally or by iv are extremely potent in inducing GH secretion o mediated in part by an increase in GHRH output and a decrease in SST secretion o mediated in part by conversion of arginine to nitric oxide, a gaseous neurotransmitter involved in the hypothalamic control of the anterior pituitary function nonesterified fatty acids reduce GH secretion without known mechanism Exercise and Stress nonspecific but potent stimulators of GH secretion stimulate GH secretion

Growth Hormone Receptor belongs to cytokine/hematopoietin receptor superfamily has 3 domains o extracellular domain o single transmembrane domain o cytoplasmic domain activates Jak/STAT pathway

Function of GH Growth & Metabolism stimulates proliferation of the cartilage cells in the epiphyseal plates at the ends of long bones, this leads to growth as ossification occurs at diaphyseal and epiphyseal ossification centers increase DNA, RNA, and protein synthesis (anabolite synthesis) promotes utilization of fat decreases utilization of carbs and decreases sensitivity to the plasma glucose- lowering action of insulin, increase blood sugar levels

stimulates uptake of AA by muscle and liver

Insulin-Like Growth factors (IGFs) in study done with poodles body size was found to parallel level of IGF results showed that body weight is correlated with IGF levels rather than with the GH secreting capacity thus providing indirect evidence for IGF-I as an important in vivo growth promoting factor IGFs are small peptides Include IGF-I and IGF-II They belong to a family of peptide hormones that include relaxin and insulin They share a degree of structural similarity with insulin Circulating IGFs are mainly produced in the liver, also produced locally in many tissues They circulate in the plasma bound to IGF binding protein GH stimulates the production of IGFs and IGFBPs in the liver Functions: growth and metabolism o IGF-I: Knockout have slow growth in the uterus and very poor growth after birth (growth factor for all stages) o IGF-II: knockouts shows slow growth in uterus, near normal growth after birth (fetal growth factor) IGF Receptors: Type I & Type II o Type I: similar to the insulin receptor (tyrosine kinase) Binds IGF I > IGF II Knockouts have poor growth in the uterus and after birth This receptor is likely the major signaling receptor of IGFs o Type II: has different structure and function than insulin receptor Binds IGF II May function to destroy excess IGF II in the fetus Knockout fetuses are larger than the controls

Assessment of GH Levels

GH Deficiency : provocative tests Screening Tests o Exercise o 90 mins after onset Definitive Tests o Insulin induced hypoglycemia o L-DOPA o Clonidine o Arginine

GH Excess: provocative tests Glucose Load Test normally glucose suppresses GH secretion, however in patients with GH excess GH levels are not suppressed 30-90 mins after following an oral glucose load IGF I measurement very sensitive GH Axis Diseases Hypopituitarism Pituitary hormone deficiency Caused by hypothalamic or pituitary disease including tumors, inactivating mutations of releasing hormones or their receptors McCune-Albright Syndrome Mutation of G protein that causes decreases GTPase activity, constituitive adenylyl cyclase activity in one or more endocrine glands Can cause acromegaly, precocious puberty, hypothyroidism, Cushings disease, bone dysplasia etc. Short Stature/Pituitary Dwarfism GH deficiency, many possible causes, pituitary tumor, genetic, etc. Short Stature/Laron Dwarfism GH resistance, GH receptor defects, GH receptor deficiency Caused by mutations or deletions in the GH receptor gene With elevated GH levels and depressed IGF and IGFBP levels The most severe IGF deficiency

Gigantism Results from GH excess early in life The bones do not mature and growth plates in bones continue to grow Overgrowth of long bones, CT, and visceral organs Symmetrical enlargement of the body Often a pituitary hormone secreting excess GH Often loss of other AP hormones Acromegaly Results from excess GH in adults Long bones are already fused and elongation of the long bones is no longer possible Therefore see a overgrowth of certain bones, overgrowth of CT, and overgrowth of visceral organs

Both gigantism and acromegaly are caused by eosinophilic adenomas consisting of somatotropes leading to excessive GH secretion

GHRH-R Gene Mutation Some patients with short stature and GH deficiency have mutated GHRHR genes How to treat GH deficiency? o Past: pituitary extracts from cadavers o Now: recombinant GH protein IGF-I Deficiency Example: 15 yr. old with history of pre and post-natal growth failure Sensoneural hearing loss and mental retardation Patient had homozygous deletion of exons 4 and 5 of IGF-I gene IGF-II Excess Developmental abnormalities Beckwith- Wiedemann Syndrome in humans o Overall overgrowth o Organ overgrowth o Skeletal abnormalities o Increased risk of tumors IGF-I Receptor Deficiency Results in intrauterine and post-natal growth retardation

9/6/12

Pituitary (3)
Prolactin A protein hormone with 199 AA A member of the GH and PRL superfamily In humans, structurally related to GH (they are evolved from gene duplication of a common ancestral gene)

Produced by lactotrophs which constitute up to 25% of the population of anterior pituitary cells. Both number and size of the lactotrophs are increased by estrogen, particularly noticeable during pregnancy Serum PRL levels are higher in nonpregnant women than in men reflecting the difference in estrogen production between the two sexes The circulating PRL levels are increased during sleep in both males and females Expressed in pituitary, placenta, and immune system PRL Releasing Peptide (PRP) specifically stimulates PRL release PRP receptor is a G protein coupled receptor expressed in lactotrophs PRL is the only anterior pituitary hormone with predominant negative control by the hypothalamus which is mediated by dopamine Dopamine causes repression of PRL and is the main control mechanism Dopamine receptors in the pituitary act by inhibiting the cAMP pathway

Roles of PRL Induces mammary gland development Stimulates lactation Stimulates lymphocyte proliferation Ovary regulate luteum status Gonads regulates steroid production Prolactin Axis Disease Prolactinoma Benign tumor of the pituitary gland that produces prolactin, it is the most common type of pituitary tumor Symptoms are caused by too much PRL in the blood (hyperprolactinemia) or by pressure of the tumor on surrounding tissues Autopsy studies indicate that 25% of the U. S. population have small pituitary tumors, 40% of these produce PRL but most are not considered clinically significant; clinically significant tumors affect the health of approximately 14 out of 100,000 people The cause of pituitary tumors is poorly understood and most are sporadic, they are not genetically passed from parents to offspring In women high blood levels of PRL often cause infertility and changes in menstruation and in some women periods may disappear altogether Women who are not pregnant or nursing may begin producing breast milk, some women may experience a los of libido and intercourse may become painful due to vaginal dryness

In men the most common symptom of prolactinoma is impotence and because men have no reliable indicator such as menstruation to signal a problem many men delay going to the doctor until they have headaches or eye problems caused by the enlarged pituitary pressing against nearby eye nerves, they may not recognize a gradual loss of sexual function or libido, only after treatment do some men realize they had a problem with sexual function Treatment: o Dopamine agonists o Remove surgically o Radiation treatment

Parkinsons Disease and PRL Dopamine is a negative regulator of PRL Since Parkinsons has a decreased amount of dopamine you would expect an increase in PRL But there is not elevated PRL found in PD But can see effects on PRL when PD patients treated with systemic drugs that act on the dopamine system

Thyroid Stimulating Hormone (TSH) Secreted by thyrotrophs that make up to 15% of the cells of AP One of the 3 pituitary glycoprotein hormones (TH, LH, FSH) All have a common alpha subunit (alpha glycoprotein subunit) All have structurally distinct beta glycoprotein subunits (e.g. TSH beta) Many disulfide bridges (5 in alpha subunit, 6 in beta unit) Sugar moieties are important to activity Roles of TSH Affects thyroid gland activity via stimulation of cAMP pathway 1. Activates iodide uptake by thyroid gland 2. Activates T3, T4 synthesis on thyroglobulin in the thyroid colloid 3. Activates TG-T3, T4 uptake into thyroid follicles 4. Activates lysosomal release of T3 and T4 from TG 5. Activates the release of T3 and T4 into blood 6. Promotes general growth of the thyroid gland Regulation of TSH by TRH TRH thyrotropin releasing hormone Produced by the hypothalamus to stimulate TSH and PRL release A tripeptide Made from a pro-TRH precursor protein that fives five copies of the bioactive TRH peptide TRH receptor is a G protein coupled receptor Binding of TRH to TRH receptor activates IP3/Ca2+ pathway

Adrenocorticotrophic Hormone (ACTH) Secreted by corticotrope cells of the anterior pituitary Human ACTH is 39 AA peptide hormone Product of the pro-opiomelanocortin, POMC, gene Regulates the cells of the adrenal gland cortex which are steroidogenic and secrete steroid hormones that affect carbohydrate and mineral metabolism Adrenal cortex makes aldosterone, cortisol and corticosterone

Mechanism of Action ACTH receptor found in the adrenal cortex ACTH receptor belongs to the melanocyte stimulating hormone receptor (MSH-R) family G protein coupled receptor Activates cAMP pathways in target cells Regulation of CTH by CRF Corticotrophin Releasing Factor CRF released by the PVN of the hypothalamus 41 AA peptide hormone stimulates ACTH release from the anterior pituitary synergizes with AVP to promote ACTH release can also stimulate MSH release CRF Receptors 7 transmembrane helix receptor activates cAMP pathways found in pituitary, hypothalamus, and some other tissues

HP Axis Diseases

ACTH Excess excess ACTH overproduction of cortisol symptoms = obesity, fatigue, high blood pressure, bruising, female menstrual problems, moon face and buffalo hump can be caused by pituitary adenomas that secrete ACTH (= Cushings Disease ) or by either tumors that secrete ACTH (in the ) or by adrenal tumors that secrete cortisol treat by removing the tumor Addisons Disease typified by low cortisol and pathologic alterations in cellular metabolism caused by adrenal failure (primary A.D.) like due to TB or can be caused by insufficient ACTH (secondary A.D.) symptoms = low bp, weakness, headaches, fainting, fever, and pain in primary A.D., can have a high ACTH because no negative feedback at AP sometimes results in hyperpigmentation rare 6/100,000 Luteinizing Hormone & Follicle-Stimulating Hormone the HPG axis: hypothalamus pituitary gonads FSH and LH are gonadotrophs Made in gonadotrope cells of the AP Glycoprotein dimer hormones (like TSH), alpha subunit is identical in the three hormones; three different beat subunits confer biological specificity to each hormone

Functions LH FSH in females induces ovulation in female initiates steroidogenesis on ovarian follicles in male stimulates Leydig cells to produce testosterone half life = 60 mins in females stimulates development of ovarian follicles in females stimulates secretion of estrogen in males stimulates spermatogenesis in males stimulates sex hormone binding globulin half life = 170 mins

Control of the gonadotropins GnRH Gonadotropin Releasing Hormone: GNRH decapeptide hormone derived from 92 AA precursor made in the hypothalamus GnRH released periodically (90 min interval) most important regulator of LH and FSH stimulates the release of both LF and FSH from the pituitary inject anti-GnRH antibodies o FSH and LH lower in both sexes o In females ovulation stops o In males testicles atrophy

Pro-GnRH transferred to Golgi apparatus Cleaved to GNRH and GAP Packaged into storage vesicles Released in response to neurotransmitter by exocytosis Vesicle guided to membrane along microtubules Fuses with membrane Membrane lyses and GnRH released and travels via portal system to AP

GnRH-R Associated Diseases GnRH-R gene mutation cause hypogonadotropic hypogonadism in humans Delayed or absent sexual development Mutant receptors are misfolded and are often not efficiently targeted to the cell membrane Puberty The beginning of sexual maturity, the period when a child changes physically, hormonally, and sexually and becomes able to reproduce Precocious Puberty Secondary development occurs before the age of 9 years in boys or 8 years in girls Early but normal sequence of pubertal development events Causes often unknown True (Central) Precocious Puberty o Emanates from premature activation of the hypothalamic pituitary-gonadal axis (GnRH dependent) o Treatment: antagonist of GnRH Pseudo (Incomplete) Precocious Puberty o Peripheral hypersecretion of estrogen or androgens

o The hypothalamic pituitary-gonadal axis is not involved (GnRH independent) o Treatment: inhibitors of estrogen or testosterone synthesis or effect Delayed Puberty Puberty is delayed when there is no sign of pubertal development by the age of 13 in years old in girls and 14 years in boys Hypogonadotropic hypogonadism: gonadotropin or GnRH therapy 9/11/12

Pituitary (4)
Intermediate Pituitary Also known as the intermediate lobe of the pituitary or pars intermedia, a part of adenohypophysis Contains a cell population called melanotropes Intermediate lobe is not distinct in adult humans Produces MSH MSH Also called alpha melanotropin A 13 AA melanotropic peptide Produced by melanotropes in the intermediate pituitary Intermediate lobe is not distinct in adult humans but alpha MSH may be produced in AP Also found in the arcuate nucleus in the hypothalamus Alpha-MSH derived from proteolytic processing of ACTH (also generates CLIP peptide whose role if it has one is unknown) ACTH, alpha-MSH, beta-MSH and beta LPH are all POMC derived peptides with melanotropic activity

MSH Receptor MSH Mechanism MSH receptors found in melanocytes, adrenal and brain Five MSH receptors have been identified: MSH-R1,3,4 and 5 bind alpha MSH, MSH-R2 only bings ACTH G protein coupled receptor Activate the cAMP pathway

Melanin Concentration Hormone 19 AA peptide in frogs an d mammals MCH may have MSH-like activity Function of MSH melanin controls pigmentation in the skin of many vertebrates melanocytes or melanophores contain the colored polymer melanin which accumulates in melanosomes. Melanosomes can be concentrated in the or dispersed within the cell or secreted into surrounding cells size of the intermediate pituitary gland is correlated with the ability to change color, lizards large pituitary other roles in humans may include: o behavior o anti-inflammatory o pregnancy Control of MSH daylight perceived and signal from pineal gland sent to pituitary to regulate alpha-MSH secretion predominantly negative control mediated by dopamine like PRL dopamine receptor antagonists (such as chlorpromazine) promote MSH secretion dopamine agonist (such as argot alkaloid) inhibit MSH secretion

Pituitary Development

Pituitary transcription factors some of these are pituitary specific (Pit-1) some become pituitary specific (Lhx3) some are pituitary- enriched some are not pituitary enriched but act in combination with other factors Primary Transcription factors - Pit-1 a transcription factor containing a POU homeodomain (POU homeodomain transcription factor) POU homeodomain transcription factors are a family of transcription factors characterized by the presence of a bipartite DNA-binding domain known as the POU domain. The POU domain contains two subdomains, a POU specific domain and a POU homeodomain The POU domain was originally identified as a region of approximately 150 amino acids shared between the Pit-1, Oct1 &2 and Unc-86 transcription factors Anterior pituitary specific transcription factor expressed in somatotropes, lactotropes, and thyrotropes activates the GH, PRL and TSHb gene animals with mutations in their Pit-1 genes are dwarfed and lack these three pituitary cell type and their hormones humans with mutations in their PIT-1 genes are of short stature and have pituitary disease (missing GH + PRL + TSH = Combined Pituitary Hormone Deficiency) Pit-1 Regulates Gene Expression

Pit-1 Target Genes hormone genes: GH, TSH beta, PRL receptors: GRF-R, TR beta2 Pit-1: autoregulates its own gene (cell stability) The snell mouse has a point mutation in the Pit-1 gene that changes one AA in the DNA binding domain an inactivates the protein; the mutation is recessive, snell mice are dwarfed and lack G, PRL, and TSH hormones/cells

Prop-1 A paired homeodomain transcription factor characterized by a bipartite DNA binding domain shared by transcription factors PAI and RED Expressed exclusively within embryonic pituitary PROP-1 gene mutated in some home CPHD Ames Dwarf mice o Harbors a naturally occurring seine to proline substitution within the homeodomain of Prop-1, resulting in a mutant protein with an eightfold lower DNA-binding affinity than wild type Prop-1 Phenotype is similar to snell dwarfs No GH, TSH, PRL hormones/cells, but also LH and FSH deficiency No Pit-1 expression in the pituitary

Prop-1 Regulates Anterior Pituitary Development

Lhx3 A LM homeodomain transcription factor The LM domain a multi-functionl protein/protein interaction domain, was first recognized in several other members of this class of transcription factors Expressed in the developing brain Expressed in Rathkes pouch Pituitary specific in adult after birth only expressed in the pituitary Activates pituitary genes such as aGSU, PRL, Pit-1, TSH Regulates anterior pituitary development

Lhx3 Regulates Anterior Pituitary Development Mice with deleted Lhx3 genes have no anterior and intermediate pituitary, mice die at birth Humans with mutated Lhx3 genes have no GH, PRL, TSH, FSH and LH . they do have ACTH

9/13/12

The Adrenal Glands

the adrenal cortex is derived from mesoderm and comprises 90% of the adrenal gland, contains three zones that each make different steroid hormones and are subjected to different regulation the adrenal medulla arises from neuroectoderm, comprises 10% of the adrenal gland, contains chromaffin cells that make catecholamine, also contains nerve fibers and medullary arter and vein

Aldosterone (mineralocorticoid) Cortisol (glucocorticoid) DHEA: dehydroepiandrosterone (androgen precursor) Adrenal Cortex Cells secrete steroid hormones rich in lipid droplets Glomerulosa zone: ovoid groups of cuboidal cells, secretes aldosterone Fasciculata zone: cords of cells with spongy appearance, secretes cortisol Reticularis zone: interconnecting network of cels sinusoidal blood vessels, secrete DEHA and its sulfated derivatives Adrenal Medulla Cells Secrete catecholamine hormones: the adrenaline surge Chromaffin cells (uptakes chromium, oxidized by potassium dichromate to a brown color) are the endocrine cells, they secrete either adrenaline or noradrenaline Each chromaffin cell is innervated by a sympathetic neuron Rich in catecholamine hormone secretory granules Can consider the adrenal medulla a modified ganglion

The Adrenal Medulla- Catecholamines Catecholamine Biosynthesis

Adrenal medullary epi makes up only 10% of circulating catecholamine thus most of it circulating is from extra adrenal sources

Epinephrine/Norepinephrine Signaling o G proten coupled receptors o Several subtypes -adrenergic receptors (1 and 2) -adrenergic receptors (1, 2, and 3) o Both epinephrine and norepinephrine activate both - and adrenergic receptors. The response of a given system is determined by the relative expression levels of the various adrenergic receptors.

Effects of Catecholamine Arousal Alerting Pupillary dilation Piloerection Sweating Bronchial dilation Tachycardia Constriction of sphincters etc. Metabolic Actions Stimulate glycogenolysis, increasing blood sugar level. Stimulate lipolysis, increasing blood free fatty acid level. Epi and NE Effects

Clinical Use of Adrenergic Receptor Agonists and Antagonists For example: Disease Cause Use of drug Mechanism Asthma bronchocontraction -agonist bronchodilation Hypertension cardioacceleration -antagonist decrease CO -adrenergic receptor agonist = isoproterenol -beta adrenergic receptor antagonist (-blocker) = propranolol Catecholamine Metabolism

Catecholamine Excess Pheochromocytomas: Benign tumor arising from the adrenal medulla. Clinical features: effects of epinephrine and norepinephrine excess Nervousness Sweating Tremor

Hypertension Tachycardia Palpitation Hyperglycemia etc. 9/18/12

The Adrenals (2)

The adrenal cortex steroid hormones

first ACTH stimulate the proliferation of the cells then it stimulates production of the cells in these two layers Choleste rol

HDL-

CYP11 A

cholesterol serves at the precurson molecue in the biosynthesis of steroids the adrenal gland can produce some cholesterol of its own but it mainly draws the cholesterol from the blod the HDL-R and LDL-R function as transporters that place the cholesterol from the blood into the cytsosol the rate limiting step is the transfer of cholesterol from the outer mitochondrial membrane to the inner mitochondrial membrane which is mediated by StAR (if a problem exist in this then the whole process of steroid synthesis is blocked) the CYP11A (cholesterol side chain cleavage enzyme) resides in the IMM the CYP11A converts cholesterol intro pregnenolone, the first step of structural modification of cholesterol in the biosynthesis of all adrenal steroids the firs step take place in the mitochondria and the rest of the steps take place in the ER a process of sequential and enzymatic modifications of cholesterol structure most of the enzymes are hydroxylases and dehydrogenases zone specific expression of cytochrome P45 enzymes determines the zone specific steroidogenesis

Cortisol - major glucocorticoid in humans essential - essential for life - maintains critical expression during prolonged stress - most effects are permissive Cortisol Release - episodic and variable - pulses follow ACTH pulses by 15-30 mins - then get negative feedback - burst just before you wake in the morning Circulation - 85% bound to transcortin (GC binding alpha 2-globulin, corticosteroid) in circulation - 15% bound to albumin in circulation - 5% free in circulation - half life is 70-90 mins, for hormone very long

cortisol binds to the glucocorticoid receptor and when there is noting bound to the GR heat shock protein are bound but when cortis binds heat shock protein dissociates, the receptor undergoes phosphorylation, nuclear translocation and dimerization, then binds to glucocorticoid response elements the glucocorticoid response element is an inverted repeat synthetic cortisols with greater selectively are widely used clinically in the treatment of diseases in which the inflammatory reactions are harmful (rheumatoid arthritis) or in which there is a need to block the immune response (prevention of rejecting transplant)

Aldosterone - the primary mineralocorticoid - arises from the zona glomerulosa of the adrenal cortex - circulates freely in the blood - half life is 15-20 mins - metabolized predominantly in the liver and excreted in the urine - the major function is to control body fluid volume by increasing sodium reabsorption in the kidneys

Regulation of Aldosterone Synthesis and Secretion - aldosterone synthesis and secretion is regulated principally by angiotensin II and serum potassium with lesser effects of ACTH

ACT H

Regulation of Aldosterone Synthesis and Secretion - renin is a proteolytic enzyme - renin cleaves angiotensin in the blood - angiotensin and angiotensin I are inactive - angiotensin II is biologically active - angiotensin converting enzyme (convert angiotensin I to angiotensin II) is found in the plasma membrane of vascular endothelial cells throughout the body - angiotensin II exerts two major actions: direct arteriolar vasoconstriction and stimulation of aldosterone synthesis and secretion - angiotensin II is the primary stimulus to aldosterone synthesis and secretion - juxtaglomerular cells synthesize and secrete proteolytic enzyme renin Renin/Angiotensin/Aldosterone a kidney adrenal axis drop in blood volume/pressure macula densa sodium levels change , transmural pressure changes, renal nerve signal juxtaglomerular cells of glomerulus produce renin hormone renin acts on angiotensin, leading to production of angiotensin (I and II) angiotensin II promotes aldosterone production by adrenal aldosterone causes sodium influs in the kindey, leading to a rise in blood volume angiotensin II also stimulates ADH release, leading to blood volume rising angiotensin II also stimulates CO and increase peripheral resistnace (vasoconstriction) leading to rise in bp rise in blood volume/pressure

Mineralocorticoid Receptor (MR) - The mineralocorticoid receptor (MR) binds aldosterone and glucocorticoids with equal affinity. - In aldosterone target tissues, like epithelial cells of the distal colon and the principal cells of the collecting ducts in the kidney, the MR is protected from glucocorticoids by the action of the enzyme 11betahydroxysteroid-dehydrogenase type 2 (11betaOHSD2), allowing aldosterone to specifically activate the receptor. However, in MRexpressing cells, which lack 11betaOHSD2, like the neurons of the limbic system in the brain, MR is mainly activated by glucocorticoids. Adrenal Androgens - DHEA and its sulfated derivative DHEA-S - They are androgen precursors (not biologically active and do not activate androgen receptor). - They can be converted to the potent androgen testosterone in peripheral tissue. - Fetal adrenal gland produces vast amounts of DHEA-S in uterus, which are converted into estrogen by the placenta.

After birth, DHEA-S production falls dramatically. They are not essential for life. DHEA is available in USA as a nutritional supplement that may be taken without a physicians advice. The precise physiological roles of adrenal androgens and the mechanisms that regulate their production remain poorly defined.

Adrenal Related Diseases Cushing's Syndrome - Overproduction of cortisol - Symptoms = obesity, fatigue, high blood pressure, bruising,female menstrual problems, moon face, buffalo hump - Can be caused by pituitary adenomas that secrete excessive ACTH (secondary Cushings syndrome) - Or by other tumors that secrete ACTH - Or by adrenal tumors that secrete excessive cortisol (primary Cushings syndrome) Cushing's Disease Caused by pituitary adenomas that secrete excessive ACTH (secondary Cusings syndrome). Addison's Disease (rare) Insufficient production of cortisol Adrenal failure (primary AD) due to tuberculosis (TB), fungal infections, autoimmune disease, etc. can get high ACTH (no negative feedback at the AP), hyperpigmentation Or caused by insufficient ACTH (secondary AD) Symptoms = low blood pressure, fainting, etc.

Hyperaldosteroidism - Conn's Disease (adrenal tumor secretes aldosterone) - Or can be 17 alpha-hydroxylase defect - causes overproduction of aldosterone Congenital Adrenal Lipoidal hyperplasia - StAR protein mutated - Adrenal and gonadal steroid insufficiency - Salt loss

Congenital Adrenal Hyperplasia - 21-hydroxylase defects - Low cortisol, no feedback, high ACTH, adrenal hyperplasia - In F get virilization (hair incr.), sex organ change, misassignment of sex due to high androgens - In M get early maturation of genitalia and growth spurts

Mineralocorticoid receptor knockout mice: pathophysiology of Na+ metabolism - Mineralocorticoid receptor (MR)-deficient mice were generated by gene targeting. - Animals had a normal prenatal development. - During first week, MR (-/-) mice developed symptoms of pseudohypoaldosteronism. - At day 8, MR (-/-) mice showed hyperkalemia, hyponatremia, and a strong increase in - renin, angiotensin II, and aldosterone plasma concentrations. - Lost weight and died at around day 10 from dehydration by renal sodium and water loss. - The fractional renal Na+ excretion was elevated >8-fold. - MR-deficient neonates die because they are not able to compensate renal Na+ loss - Daily subcutaneous injections of isotonic NaCl solution until weaning and continued oral NaCl supply lead to survival of the MR knockout mice; however, the renal salt-losing defect persists.

Pineal Gland
The pineal is a pea-sized, pine-cone shaped gland in the brain McCord & Allen (1917) added bovine pineal extract to pond water color of tadpoles' skin lightened (melanosome aggregation)

Frogs and fish Pineal is a photoreceptor (3rd eye) Sends direct signals to the brain Birds Pineal is a photoendocrine transducer Indirect effect Direct light pineal hormones Mammals Pineal is a neuroendocrine transducer Eye brain pineal hormones (melatonin)

Pineal Cell Types Pinealocytes = true pineal cells Astrocytes = brain support cells Nerve input fibers Pineal Tumors True pineal tumors Astrocytomas Pineal cysts Melatonin - Synthesized in pineal from the amino acid tryptophan by pineal-specific enzymes Physiological effects - regulate and entrain circadian rhythms - hypnotic effects - seasonal reproduction (animals) - thyroid physiology - retinal physiology Melatonin Signaling - Activates melatonin receptors (3 types), causing inhibition of adenylyl cyclase and decrease of cAMP level. Other effects/uses? - ageing/longevity - antioxidant properties - anticancer effects - blindness - antipsychotic effects - migraine headaches - Parkinson's disease

Melatonin Hypothesis Observations (rodents):

(Whitman & Axelrod, 1963)

1. Increase light pineal size decreases ovary bigger 2. Remove pineal ovary size increases 3. Add melatonin ovary size decreases Hypothesis: Melatonin = hormone of the pineal gland that is controlled by light signals and regulates gonadal function.