General principles of endocrinology Contact Karron J. James, Ph.D. • • • • Office: GB6, Block B Email: kjames@auamed.net Phone 484-8900 ext 1041 Office hours: Mon – Fri 9am-10am, 3pm-4pm on Microsoft Teams® OR by appointment Goal 1: To understand the general principles of endocrine physiology Learning Objectives a. List the principal endocrine organs and tissues. b. Contrast the terms endocrine, paracrine, and autocrine and give an example of each. c. Describe the secretion, transport in the circulation, cellular mechanisms of action, regulation of receptors and other characteristics of peptide, steroid, and amine (tyrosine) hormones. d. With examples, contrast the location and signalling pathways of membrane bound and intracellular hormone receptors. e. Compare and contrast hormone actions that are exerted through changes in gene expression with those exerted through changes in protein activity, such as through phosphorylation f. For membrane bound hormone receptors, describe the process of activation, inactivation, up-regulation, down-regulation, sensitization, and desensitization Readings Molina, P.E. (2018). Endocrine Physiology (5th ed.) McGraw-Hill Education http://auamed.idm.oclc.org/login?url=https://access medicine.mhmedical.com/book.aspx?bookid=2343 Chapter 1: General Principles of Endocrine Physiology • The endocrine system: physiologic functions and components • Hormone chemistry and mechanisms of action (up to and incl Fig 1-3) • Hormone receptors and signal transduction Objective 1a. List the principal endocrine organs and tissues Fig 1-1. Molina Signalling Basics • Cell communication: transduction of message from one cell to (usually) another cell resulting in altered behaviour of target cell • Basic requirements: – Signalling cell – Extracellular ligand (hormone) – Target (responding) cell – Receptor (plasma membrane or intracellular) Goal 1: To understand the general principles of endocrine physiology Learning Objectives a. List the principal endocrine organs and tissues. b. Contrast the terms endocrine, paracrine, and autocrine and give an example of each. c. Describe the secretion, transport in the circulation, cellular mechanisms of action, regulation of receptors and other characteristics of peptide, steroid, and amine (tyrosine) hormones. d. With examples, contrast the location and signalling pathways of membrane bound and intracellular hormone receptors. e. Compare and contrast hormone actions that are exerted through changes in gene expression with those exerted through changes in protein activity, such as through phosphorylation f. For membrane bound hormone receptors, describe the process of activation, inactivation, up-regulation, down-regulation, sensitization, and desensitization Obj 1b: Contrast the terms endocrine, paracrine, and autocrine and give an example of each • Signalling by secreted molecules 1. Endocrine: bloodstream 2. Paracrine: nearby 3. Autocrine: self Fig 1-3. Molina •All signalling molecules only active on target cells that express an appropriate receptor Goal 1: To understand the general principles of endocrine physiology Learning Objectives a. List the principal endocrine organs and tissues. b. Contrast the terms endocrine, paracrine, and autocrine and give an example of each. c. Describe the secretion, transport in the circulation, cellular mechanisms of action, regulation of receptors and other characteristics of peptide, steroid, and amine (tyrosine) hormones. d. With examples, contrast the location and signalling pathways of membrane bound and intracellular hormone receptors. e. Compare and contrast hormone actions that are exerted through changes in gene expression with those exerted through changes in protein activity, such as through phosphorylation f. For membrane bound hormone receptors, describe the process of activation, inactivation, up-regulation, down-regulation, sensitization, and desensitization Hormones • Regulate physiologic processes, eg. reproduction, growth, metabolism • Can be classified as peptide, steroid, amine Peptide or Protein Hormones • Inactive precursor stored in secretory vesicles near plasma membrane, released via exocytotic pathway • Bind plasma membrane receptors Major classes of plasma membrane receptors 1. 2. 3. 4. Ligand-gated ion channels GPCR Enzyme-coupled Cytokine receptors 1. Ligand-gated Ion Channel • Ionotropic receptor • Membrane protein complex that includes ligand binding sites forms a pore Ion Signaling molecule • Example? https://www.nature.com/scitable/topicpage/ion-channel-14047658 2. G protein-coupled receptor • a.k.a. 7-TM domain receptor • Relies on second messengers to relay intracellular signals • Binding phosphorylation of GDP associated with trimeric G protein complex Fig 1-5. Molina Gα proteins • Gαs – Activates adenylyl cyclase • Gαi – Inhibits adenylyl cyclase • Gαq – Activates phospholipase C • Gα12/13 – Activates RhoA GPCR—Gs signalling Fig 1-5. Molina Fig 15-33. Alberts. GPCR—Gi signalling Fig 1-5. Molina https://www.news-medical.net/whitepaper/20170525/Using-CLARIOstarc2ae-Microplate-Reader-forReal-Time-Detection-of-Gi-and-Gs-Signaling-in-Living-Cells.aspx GPCR—Gq signalling example, OXYTOCIN Fig 15-36. Alberts. Fig 1-5. Molina GPCR—G12/13 Signalling • Receptor ligands: thrombin, lysophosphatidic acid, sphingosine-1-phosphate, thromboxane A2 • Receptor activation activation of Rho GEF activation of RhoA • Effects in the cytoplasm and in nucleus Fig 1-5. Molina 3. Enzyme-coupled Receptors • Serine-threonine kinase Fig. 15-65. Alberts. • Receptor tyrosine kinase https://www.nature.com/scitable/topicpage/rtk-14050230 RTK Signalling can occur via 3 pathways • Signal transduction pathways: a. PI3-K b. Ras/MAPK (Grb/SOS) c. PLC Fig 8-1. Goodman, S.R. Medical Cell Biology. 3ed. Academic Press. a. PI3-K Pathway—insulin, IGF-1 signalling • Ligand binding RTK~P • Activated RTKs recruit IRS, and phosphorylate • formation of binding sites for PI3-K, also phosphorylated • PI3-K: PIP2 PIP3 • PIP3 recruits Akt (PKB), phosphorylates it phosphorylation cascade downstream • GLUT4 translocates to membrane. Lipid, protein synthesis, etc. Fig 1-6 https://employees.csbsju.edu/hjakubowski/classes/ch331/signaltrans/ST_9 C9_Insulin_Sig_PI3K_Ak.html b. RAS/MAPK Pathway • Activated RTKs recruit intracellular signal transducers near membrane, Eg. Shc, IRS1. Phosphorylates • Formation of binding sites for Grb2. Phosphorylates • Grb2/SOS catalyzes conversion of Ras from inactive to active (GTPbound) form • Ras-GTP interacts with and stimulates downstream effectors, leading to activation of the MAP kinases ERK1 and 2 • ERKs enter nucleus, phosphorylate and activate Tc factors for growth, differentiation Insulin (Insulin receptor) c. PLC Pathway • Activated RTKs recruit intracellular signal transducers near membrane – Bind and phosphorylate substrates w/ SH2 domains, eg. PLCγ stimulation of phospholipase activity • PLC: PIP2 IP3, DAG • IP3 Ca2+ release from ER • Ca2+, DAG activate PKC activate Tc of genes involved in proliferation, survival, etc. http://publication.letstalkacademy.com/ip3-dag-pathway-of-receptor-tyrosinekinase-in-cell-signalling Receptor Serine Threonine Kinases • When activated, cell surface receptors phosphorylate S or T on substrate proteins • One of a group of receptors that binds TGFβ family of signalling molecules • Ligand-receptor complex binds Smad proteins • Smads enter nucleus to regulate transcription of target genes Fig. 15-65. Alberts. 4. Class I Cytokine Receptors • No intrinsic TK activity • Associate with ‘non-receptor (cytosolic) tyrosine 2’—Jak2 • Jak/STAT pathway Fig 1-6 Steroid Hormones • Include adrenal (eg. cortisol), sex hormones (testosterone, estradiol, progesterone) – Adrenal cortex, gonads, placenta • Bind receptor in the cytoplasm • Ligand binding binding of DNA regulatory sequence (HRE) by complex regulation of transcription studyblue.com Steroid hormones • Derived from cholesterol • Synthesized and quickly released bloodstream, bound by carrier proteins hormone, only, taken up by target cell • Vitamin D3 Retinoic acid • Not a steroid hormone • Retinoic acid—synthesised from retinol, not cholesterol • Tc factor, important for patterning during embryogenesis – Receptor located in nucleus Amine Hormones Eg. Thyroid hormones, epinephrine*, norepinephrine* Goal 1: To understand the general principles of endocrine physiology Learning Objectives a. List the principal endocrine organs and tissues. b. Contrast the terms endocrine, paracrine, and autocrine and give an example of each. c. Describe the secretion, transport in the circulation, cellular mechanisms of action, regulation of receptors and other characteristics of peptide, steroid, and amine (tyrosine) hormones. d. With examples, contrast the location and signalling pathways of membrane bound and intracellular hormone receptors. e. Compare and contrast hormone actions that are exerted through changes in gene expression with those exerted through changes in protein activity, such as through phosphorylation f. For membrane bound hormone receptors, describe the process of activation, inactivation, up-regulation, down-regulation, sensitization, and desensitization Regulation mechanisms: See Fig 15-25. Alberts. Response to ligand-binding is regulated • Receptors may be up-regulated – May be response to reduced level of ligand – Increase transcription, translation • Decrease degradation of receptors • Activation of receptor • Receptors may be down-regulated – Reduce transcription, translation • Increase degradation of receptors Target cell can be sensitized or de-sensitized • Desensitization (adaptation) can occur after repeated or chronic exposure of receptor to hormone • Reduced response to future stimulation by same ligand or same concentration of ligand – Over time, no response to same ligand concentration – Need greater ligand concentration to produce response Fig 15-48. Alberts. • Can occur by receptor sequestration, receptor downregulation, receptor inactivation (eg. by phosphorylation) Goal 1: To understand the general principles of endocrine physiology Learning Objectives a. List the principal endocrine organs and tissues. b. Contrast the terms endocrine, paracrine, and autocrine and give an example of each. c. Describe the secretion, transport in the circulation, cellular mechanisms of action, regulation of receptors and other characteristics of peptide, steroid, and amine (tyrosine) hormones. d. With examples, contrast the location and signalling pathways of membrane bound and intracellular hormone receptors. e. Compare and contrast hormone actions that are exerted through changes in gene expression with those exerted through changes in protein activity, such as through phosphorylation f. For membrane bound hormone receptors, describe the process of activation, inactivation, up-regulation, down-regulation, sensitization, and desensitization