Hormonal regulation and circadian rhytms

1. Hormonal regulation and circadian rhytms Alice Skoumalová

2. Central and peripheral clocks in mammals Organized in a hierarchical manner • Central oscillator: • controls the behavioral rhythm • reset by external light signal • Peripheral oscillators: • slave oscillators • regulate local rhythms • reset by feeding

3. A model for feedback loops of mammalian circadian clock

4. External synchronization of the SCN to light-dark cycles

5. A circadian rhytm is 24-hour cycle in the physiological processes of living beings: • Circadian rhytms are endogenously generated; modulated by external signals (day/night rhytm) • Circadian rhytms - brain wave activity, hormone production, cell regeneration and other biological activities linked to this daily cycle Regulation of a circadian rhytm 1. The information about illumination from the retina (retinal ganglion cells with melanopsin); the retinohypothalamic tract 2. The circadian "clock" in the suprachiasmatic nucleus (SCN) 3. The SCN interprets the information on day lenght and passes it on to the pineal gland which then secretes the hormone melatonin (secretion of melatonin peaks at night) Melatonin participates in various physiological functions according to 24-hour cycle

6. Melatonin: • Ubiquitously in nature (one of the most phylogenetically ancient of all biological signaling mechanisms) • A potent antioxidant (its primary function) Melatonin is involved in various physiological functions: Circadian rhytm regulation, sleep propensity, control of sleep/wake rhytm, blood pressure regulation, immune function, retinal functions, detoxification of free radicals, control of tumor growth, bone protection, the regulation of bicarbonate secretion in the GI tract Melatonin is primarily secreted by the pineal gland; synthesis also occurs in other cells and organs Melatonin secretion is synchronized to the light/dark cycle, with a nocturnal maximum Exogenous administration Circadian rhytm sleep disorders (CRSD), insomnia, cancer, neurodegenerative diseases, disorders of the immune function, oxidative damage

7. Melatonin in plants • in high concentrations (protection from oxidative damage) • many plants represent an excellent dietary source of melatonin as an antioxidant nutrient The majority of herbs used in traditional Chinese medicine for retarding age-related changes and for treating diseases associated with the generation of free radicals contain the highest levels of melatonin

8. Melatonin synthesis The pineal gland The retina Lymphocytes The GI tract Bone marrow cells Platelets Skin Serotonin N transferase: the rate-limiting enzyme

9. Pineal melatonin production exhibits a circadian rhytm • low level during daytime, high level during night The retinohypothalamic tract: the regulation of pineal melatonin biosynthesis The retina→The suprachiasmatic nucleus (SCN) →The spinal cord (intermediolateral horn cells) →The superior cervical ganglion→The pineal gland

10. Melatonin metabolism • Melatonin is not stored; diffuses out into the capillary blood and cerebrospinal fluid • In the liver: melatonin is first hydroxylated to 6-hydroxymelatonin (by cytochrome P450 mono-oxygenases) and conjugated with sulfate • In the pineal gland and the retina: melatonin can be deacetylated to 5-methoxytryptamine (by melatonin-deacetylating enzymes)

11. Melatonin can be metabolized nonenzymatically: In all cells: It is converted into cyclic 3-hydroxymelatonin when it directly scavenges two hydroxyl radicals (OH•) In the brain: a substantial fraction of melatonin is metabolized to kynuramine derivates (the kynuric pathway) The kynuramine derivates- the antioxidant and anti-inflammatory properties 1. N-acetyl-N-formyl-5-methoxykynuramine (AFMK) 2. N-acetyl-5-methoxykynuramine (AMK)

12. Melatonin exerts actions in almost every cell in the body • Melatonin receptors-receptor mediated effects • Melatonin diffuses through membranes easily-receptor independent effects

13. Melatonin produces effects in: Receptor-independent: 1. Free radical scavenging properties 2. Inhibition of calmodulin -an attenuation of cAMP-dependent signaling cascades -decrease ER binding and activation of ERE-containing genes 3. Regulation of the quinone reductase 2 activity Receptor-dependent: 1. G-protein coupled receptors (MT1 or MT2) - an attenuation of cAMP-dependent signaling cascades - inhibition of ER 2. The mitogen activated protein kinase cascade (MEK/ERK)

14. Melatonin Scavenging of reactive oxygen species (ROS), reactive nitrogen species (RNS) and organic radicals MT1, MT2 Circadian pacemaker: suprachiasmatic nucleus CNS: antiexcitatory effects, avoidance of Ca2+ overload Quinone reductase 2 (MT3) Seasonal breeding (hypothalamus and other organs relevant to reproduction) Cytoskeletal effects: binding to calmodulin, activation of protein kinase C Elimination of toxic quinones Immune system (B cells, T cells, NK cells, thymocytes, bone marrow) Upregulation of antioxidant and downregulation of prooxidant enzymes Direct inhibition of mitochondrial permeability transition pore opening Vasomotor control: constriction via MT1 dilation via MT2 Attenuation of mitochondrial electron leakage Prevention of apoptosis Decrease of free radicals and other oxidants

15. Melatonin as an antioxidant More efficient than other antioxidants; devoid of pro-oxidant side-effects 1. Scavenging of free radicals 2. Up-regulation of antioxidant enzymes 3. Direct inhibition of free radical formation • Neurodegenerative diseases (Alzheimer‘s, Parkinson‘s and Huntington‘s disease) - potential role of melatonin

16. Melatonin as an oncostatic substance Melatonin inhibits the carcinogenesis 1. Antioxidant activity 2. Modulation of the estrogen signalling pathway • Melatonin has demonstrated oncostatic effects against a variety of tumor cells: estrogen-positive breast cancer cell lines, ovarian carcinoma cell lines, endometrial carcinoma, human uveal melanoma cells, prostate tumor cells, intestinal tumors • Melatonin ameliorates side effects of antitumoral therapeutic regimens (myelotoxicity, lymphocytopenia)

17. Melatonin‘s immunomodulatory function Melatonin has an immunomodulatory role 1. Enhances the production of cytokines 2. Antiapoptotic and antioxidant actions 3. Direct effect on the regulation of the immune system (via receptors) • Inhibition of melatonin synthesis results in the attenuation of cellular and humoral responses: exogenous melatonin counteracts immunodeficiences • The role of melatonin in the pathogenesis of autoimmune diseases: the increased prevalence of auto-immune diseases at winter (long nights, increased levels of melatonin)

18. Melatonin as a hypnotic Melatonin promotes sleep in healthy humans MT1 receptors in SCN • The treatment of insomnia (particularly in individuals with melatonin deficiency)

19. Melatonin as a chronobiotic molecule Melatonin acts as an endogenouos synchronizer of bodily rhythms MT2 receptors in SCN • Implication in circadian rhythm sleep disorders 1. Shift-work disorder 2. Jet lag syndrom 3. Delayed sleep phase syndrome 4. Circadian rhythm disruption with ageing

20. Melatonin in depression Altered levels and phase-shift of melatonin in depressed patients MT1 receptors in the brain • Melatonin has the potential value of being used as a therapeutic agent in the treatment of mood disorders Agomelatine (a melatonin agonist) has been tested in clinical studies as a novel melatonergic antidepressant

21. GI melatonin Melatonin is synthesized in the enterochromaffin cells of the GI tract and can be released to the circulation Functions in the GIT: 1. Increase duodenal mucosal secretion of bicarbonate - duodenal protection against gastric acid 2. High concentration in the bile-prevents oxidative damage to the intestinal epithelium caused by bile acids 3. Gastroprotective efficacy: as an antioxidant

22. Melatonin in cardiovascular diseases Melatonin reduces blood pressure in hypertensive patients 1. Peripheral mechanism: vasodilatation via MT2 vasoconstriction via MT1 (cerebral vessels) 2. Central mechanism

23. Melatonin effects on bone Melatonin causes inhibition of bone resorption (protects bone during treatment with gluococorticoids that affect bone remodeling and cause osteoporosis) 1. Down-regulation of osteoclast activation 2. Direct inhibition of osteoclast function (antioxidant)

24. Melatonin in sexual maturation and in reproduction 1. Via MT receptors 2. Via nuclear receptors • Melatonin inhibits the hypothalamic-pituitary-gonadal axis (important for sexual maturation) The decline in melatonin concentration is very important for the initiation of puberty • Melatonin down-regulates GnRH genein a cyclicyl pattern over a 24-h period; the pulsatile secretion of GnRH controls LH a FSH

25. Summary • Melatonin is distributed widely in nature; it acts as a photoperiod messenger molecule, transducing photoperiod changes to various cyclic function in organism (reproduction, sleep-wake rhythms) • Melatonin is very important antioxidant (primary function in evolution) • Melatonin influences various cell mechanisms via receptors (plasmatic, nuclear) • Melatonin play a role in many pathological states: neurodegenerative disorders, circadian rhythm sleep disorders, depression, cardiovascular diseases, tumor growth, immune pathologies, bone resorption (potential therapeutic agent)