1212569_21823227.jpg logo_mu_cerne.gif 1212570_28446780.jpg logo_mu_cerne.gif Luděk Bláha, PřF MU, RECETOX www.recetox.cz BIOMARKERS AND TOXICITY MECHANISMS 08 – Mechanisms Signalling and regulation OPVK_MU_stred_2 1212569_21823227.jpg logo_mu_cerne.gif Cell communication & regulation: a target for toxicants … especially sensitively regulated processes are highly susceptible to toxicants à toxicity to REGULATIONS & SIGNALLING Hierarchy in signalling - systems: neuronal ßà endocrine - cell-to-cell hormonal & neuronal signal transmission contact channels - intracellular signal transduction 1212569_21823227.jpg logo_mu_cerne.gif Cell communication & regulation: a target for toxicants C:\Documents and Settings\Ludek Blaha\Obrázky\5.jpg 1212569_21823227.jpg logo_mu_cerne.gif Cell communication (1) C:\Documents and Settings\Ludek Blaha\Obrázky\1.jpg 1212569_21823227.jpg logo_mu_cerne.gif C:\Documents and Settings\Ludek Blaha\Obrázky\2.jpg Cell communication (2) 1212569_21823227.jpg logo_mu_cerne.gif C:\Documents and Settings\Ludek Blaha\Obrázky\3.jpg C:\Documents and Settings\Ludek Blaha\Obrázky\4.jpg Cell communication (3) 1212569_21823227.jpg logo_mu_cerne.gif Endocrine system: 1. Pineal gland, 2. Pituitary gland, 3. Thyroid gland, 4. Thymus, 5. Adrenal gland, 6. Pancreas, 7. Ovary, 8.Testis Example: feedback loop 1212569_21823227.jpg logo_mu_cerne.gif * stimulation or inhibition of growth * mood swings * induction or suppression of apoptosis (programmed cell death) * activation or inhibition of the immune system * regulation of metabolism * preparation for fighting, fleeing, mating … * preparation for a new phase of life (puberty, caring for offspring, and menopause) * control of the reproductive cycle …. etc. FUNCTIONS OF HORMONES 1212569_21823227.jpg logo_mu_cerne.gif FATE OF HORMONES: target for toxicants Toxic compounds can affect “hormone signalling” at various levels (highligted): 1. Biosynthesis of a particular hormone in a particular tissue 2. Storage and secretion of the hormone 3. Transport of the hormone to the target cell(s) 4. Recognition of the hormone by an associated cell membrane or intracellular receptor protein. 5. Relay and amplification of the received hormonal signal via a signal transduction process -> cellular response. 6. The reaction of the target cells is recognized by the original hormone-producing cells (negative feedback loop) 7. Degradation and metabolism of the hormone System regulation = HORMONES & ENDOCRINE SYSTEM 1212569_21823227.jpg logo_mu_cerne.gif ED & EDCs (endocrine disrupting compounds) = major problem in environmental toxicology Effects at all levels of hormonal action have been demonstrated à synthesis, transport, site of action …. - Multiple effects due to ED (! Not only „xenoestrogenicity“ & feminization) à immunotoxicity, developmental toxicity (ED - WILL ALSO BE DISCUSSED FURTHER) Example of ED - Intersex roach testis containing both oocytes and spermatozoa, caused by exposure to environmental oestrogens Toxicity to hormone regulation = ENDOCRINE DISRUPTION 1212569_21823227.jpg logo_mu_cerne.gif Amine-derived hormones structure: derivatives of the amino acids tyrosine and tryptophan. Examples - catecholamines and thyroxine. (small molecules - similar to organic toxicants à TOXIC EFFECTS) Adrenalin Thyroxin Types of hormones in vertebrates 1212569_21823227.jpg logo_mu_cerne.gif Peptide hormones structure: chains of amino acids. - small peptides: TRH and vasopressin; - large proteins: insulin, growth hormone, luteinizing hormone, follicle-stimulating hormone and thyroid-stimulating hormone etc. Large molecules; receptors on surfaces of the cells (Interactions with toxic chemicals less likely) Example - insulin Types of hormones in vertebrates 1212569_21823227.jpg logo_mu_cerne.gif Lipid derived “hormones” (1) - from linoleic acid, arachidonic acid - prostaglandins Types of hormones in vertebrates 1212569_21823227.jpg logo_mu_cerne.gif Lipid derived hormones 2 - steroid hormones * Small molecules - similar to organic toxicants: à several compounds interfere with steroid hormones à toxicity !!! Derived from cholesterol Examples: testosterone, cortisol, estradiol … Types of hormones in vertebrates 1212569_21823227.jpg logo_mu_cerne.gif Intracellular signal transduction: target of toxicants •- Regulation of cell life = control of major cell functions • - metabolism • - proliferation • - differentiation • - death (apoptosis) • •- Regulation controlled by complex signalling • - "network" of general pathways • - similar in all cells / different cell-specific effects • 1212569_21823227.jpg logo_mu_cerne.gif •- Consequences of signalling disruption • - unwanted changes in „homeostatic“ rates among proliferation / differentiation / apoptosis –à cell transformation (carcinogenicity) –à embryotoxicity –à immunotoxicity –à reproduction toxicity – .... and other chronic types of toxicity Intracellular signal transduction: target of toxicants 1212569_21823227.jpg logo_mu_cerne.gif Signal transduction - principles •Two major signalling processes – protein-(de)phosphorylation • ProteinKinases - PKs, ProteinPhosphatases - PPases • - secondary messengers • cAMP / IP3, PIP2, DAG, Ca2+, AA • •Three major types of signalling –1: Membrane receptors (G-protein, kinases) à activation of protein kinase A (PKA): major messenger: cAMP – –2: Membrane receptors à activation of membrane lipases à and later proteinkinase C IP3, PIP2, DAG, Ca2+, AA – –3: Cytoplasmic (nuclear) receptors (discussed in detail in other sections) – 1212569_21823227.jpg logo_mu_cerne.gif •Membrane receptors acting as ProteinKinases G-proteins & G-protein coupled receptors - GPCRs • C:\Documents and Settings\Ludek Blaha\Obrázky\1.jpg 1212569_21823227.jpg logo_mu_cerne.gif •Signalling mechanism 1 •à Activation of adenylate cyclase à cAMP à PKA C:\Documents and Settings\Ludek Blaha\Obrázky\1.jpg 1212569_21823227.jpg logo_mu_cerne.gif ßPKA is central to a number of signalling events and following effects ßIncluding modulation of “MAPKs” 1212569_21823227.jpg logo_mu_cerne.gif Mitogen Activated Protein Kinases (MAPKs) & dependent effects C:\Documents and Settings\Ludek Blaha\Obrázky\1.jpg 1212569_21823227.jpg logo_mu_cerne.gif •Signalling mechanism 2 •Activation of Phospholipase C à release of PIPs à DAG à PKC / arachidonic acid + IP3 à activation of Ca2+ signalling C:\Documents and Settings\Ludek Blaha\Obrázky\1.jpg C:\Documents and Settings\Ludek Blaha\Obrázky\1.jpg 1212569_21823227.jpg logo_mu_cerne.gif C:\Documents and Settings\Ludek Blaha\Obrázky\1.jpg 1212569_21823227.jpg logo_mu_cerne.gif C:\Documents and Settings\Ludek Blaha\Obrázky\1.jpg Different “types” of signalling crosstalk and form networks 1212569_21823227.jpg logo_mu_cerne.gif Examples •Estrogenicity of PAHs independent on activation of estrogen receptor •PAHs àmodulation of PKs/PPases: phosphorylation events à ligand independent activation of ER • • PAHs significantly potentiate the effect of 17b-estradiol (via increased phosporylation of ER) Vondráček et al. 2002 Toxicol Sci 70(2) 193 1212569_21823227.jpg logo_mu_cerne.gif •Example - cholera toxin: •CT acts as adenylate cyclase à increasing cAMP levels à TOXICITY • 1212569_21823227.jpg logo_mu_cerne.gif •Example: Lipopolysaccharides (LPS) from cell walls • à hyperactivation of intracellular signals à immunotoxicity •