8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 1 Development of Complex Curricula for Molecular Bionics and Infobionics Programs within a consortial* framework** Consortium leader PETER PAZMANY CATHOLIC UNIVERSITY Consortium members SEMMELWEIS UNIVERSITY, DIALOG CAMPUS PUBLISHER The Project has been realised with the support of the European Union and has been co-financed by the European Social Fund *** **Molekuláris bionika és Infobionika Szakok tananyagának komplex fejlesztése konzorciumi keretben ***A projekt az Európai Unió támogatásával, az Európai Szociális Alap társfinanszírozásával valósul meg. PETER PAZMANY CATHOLIC UNIVERSITY SEMMELWEIS UNIVERSITY sote_logo.jpg dk_fejlec.gif INFOBLOKK 8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 2 Peter Pazmany Catholic University Faculty of Information Technology BASICS OF NEUROBIOLOGY NEUROTRANSMITTERS II. www.itk.ppke.hu Neurobiológia alapjai (Neurotranszmitterek II.) ZSOLT LIPOSITS Basics of Neurobiology: Neurotransmitters II. 8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 3 www.itk.ppke.hu CHARACTERISTICS OF NEUROPEPTIDES NEUROPEPTIDES ARE COMPOSED OF SHORT CHAINS OF AMINO ACIDSAND SYNTHESIZED BY NERVE CELLS THEY ARE BIOLOGICALLY ACTIVE SUBSTANCES USED AS SIGNAL MOLECULESIN INTERNEURONAL COMMUNICATION IN THE CNS, THEIR NUMBER EXCEEDS ONE HUNDRED THEY ARE CLASSIFIED INTO NEUROPEPTIDE HORMONE FAMILIES BASED UPON SIMILARITIES IN ORIGIN, STRUCTURE AND FUNCTION INVENTION OF IMMUNOCYTOCHEMISTRY AND ITS USE IN NEUROSCIENCE RESEARCH OPENED A PROGRESSIVE FIELD CALLED CHEMICAL NEUROANATOMY THE DETECTION OF NEUROPEPTIDES BY IMMUNOCYTOCHEMISTRY AND THEIR mRNAs BY IN SITU HYBRIDIZATION HISTOCHEMISTRY REVEALED AN EXTREMELY COMPLEX CHEMICAL ORGANIZATION OF THE BRAIN ELECTROPHYSIOLOGICAL AND ULTRASTRUCTURAL STUDIES HAVE SHOWN THATNEUROPEPTIDES LOCALIZED IN AXON TERMINALS WERE,INDEED,RELEASED AND THEYAFFECTED POSTSYNAPTIC NEURONS Basics of Neurobiology: Neurotransmitters II. 8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 4 www.itk.ppke.hu CHEMICAL PHENOTYPE OF THE HYPOTHALAMUS THEHYPOTHALAMICPARTOFTHEBRAIN THATREGULATESTHEENDOCRINESYSTEM ISESPECIALLYRICHINNEUROPEPTIDES THECOLOREDSYMBOLSREFERTODIFFE- RENTNEUROHORMONESANDINDICATETHEIR DIFFERENTIALEXPRESSIONINHYPOTHALAMIC NUCLEI.THERELEASINGANDRELEASEINHIBIT- INGHORMONES(LHRH,TRF,CRH,SRIF,GHRH) HAVEAHIGHFUNCTIONALSIGNIFICANCE. INADDITIONTONEUROPEPTIDES,THEHYPO- THALAMUSALSOPRODUCESCLASSICALTRANS- MITTERSINCLUDINGGABAANDDOPAMINE. Basics of Neurobiology: Neurotransmitters II. 8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 5 www.itk.ppke.hu VISUALIZATION OF NEUROPEPTIDES BY IMMUNOSTAINING B THE SECTION SHOWS THE HYPOTHALAMIC AREA DEPICTED IN TRANSPARENT YELLOW SHADING IN THE PREVIOUS SLIDE ARROWHEADS POINT TO FUSIFORM NEURONS THAT SYNTHESIZE LUTEINIZING HORMONE- RELEASING HORMONE (LHRH) BLACK LHRH DENDRITES AND AXONS ARE ALSO VISIBLE IN THE IMMUNOCYTOCHEMI- CAL PREPARATION LHRH IS A 10 AMINO ACID PEPTIDE THAT REGULATES REPRODUCTION AND REPRO- DUCTIVE BEHAVIOR IMMUNOCYTOCHEMISTRY IS A FUNCTIONAL ANATOMICAL TOOL EXPLORING THE STRUC- TURAL AND BIOCHEMICAL CHARACTERIS- TICS OF NEURONS SIMULTANEOUSLY Basics of Neurobiology: Neurotransmitters II. 8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 6 www.itk.ppke.hu PROCESSING OF NEUROPEPTIDES PEPTIDERGIC NEURONS CONTAIN mRNAsIN THEIR CYTOPLASM THAT ENCODE THE NEUROPEPTIDE PRECURSOR PROTEINS THEIR SYNTHESIS TAKES PLACE AT ROUGH ENDOPLASMIC RETICULUM, THENTHE NEWLY SYNTHESIZED PEPTIDE IS DIRECTED TO THE GOLGI COMPLEX WHEREITS MODIFICATION OCCURS AFTER INITIAL MODIFICATIONS (GLYCOSYLATION, PROTEIN FOLDING, DISULFIDE BOND FORMATION, CLEAVAGE), THE MATURING PROTEINS ARE PACKED INTOSECRETORY VESICLES THAT ARE RELEASED FROM THE TRANS GOLGI FACE THE CONSTITUTIVE TRANSPORT DIRECTS THE VESICLES TO THE AXON TERMINALS UTILIZING ANTEROGRADE TRANSPORT MECHANISMS DURING THEIR JOURNEY, FURTHER PROCESSING OF THE PROTEINS MIGHT TAKE PLACE MOST FREQUENT ACTION IS THE CLEAVAGE OF THE PRECURSOR PROTEIN BY PRO-PROTEIN CONVERTASES Basics of Neurobiology: Neurotransmitters II. 8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 7 www.itk.ppke.hu THE CLEAVAGE MAY GENERATE MULTIPLE DISTINCT PEPTIDES FROM THE PRECURSOR OR PRODUCE MULTIPLE COPIES OF THE SAME NEUROPEPTIDE MOST PEPTIDES BECOME FULLY BIOACTIVE AFTER POST-TRANSLATIONAL CLEAVAGE, OTHERS UNDERGO FURTHER MODIFICATION (CARBOXYL-AMIDATION) THE NEUROPEPTIDES ARE STORED WITHIN NEUROSECRETORY GRANULES OF AXON TERMINALS. THEIR DIAMETER IS IN THE RANGE OF 80-200 nm SOME NEUROPEPTIDES HAVE BEEN SHOWN TO BE RELEASED FROM DENDRITES AND AXON VARICOSITIESINADDITIONTOTERMINAL BOUTONS NEUROSECRETORY GRANULES MAY CONTAIN MORE THAN ONE NEUROPEPTIDE. THE CO-PACKAGED NEUROPEPTIDES VERY LIKELY UNDERGO CO-RELEASE CLASSICAL AND NON-CLASSICAL NEUROTRANSMITTERS ARE ALSO KNOWN TO BE CO-SYNTHESIZED IN PEPTIDERGIC NEURONS, INDICATING THAT CELLS MIGHT USE MULTIPLE NEUROMESSENGERS IN THEIR COMMUNICATION PROCESSING OF NEUROPEPTIDES Basics of Neurobiology: Neurotransmitters II. 8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 8 www.itk.ppke.hu CO-LOCALIZATION OF TWO NEUROPEPTIDES IN THE SAME NEUROSECRETORY GRANULE SCHEME OF AN LHRH AXON CONTAINING SECRETORY GRANULES. THE TWO BLUELINES POINT TO ANELECTRON- MICROSCOPIC IMAGE OF A SINGLENEUROSEC- RETORYGRANULE DOUBLE-LABELED FOR TWO NEUROPEPTIDES THE EM MICROGRAPH DEPICTS A NEUROSECRETORY GRANULE FROM AN LHRH NERVE TERMINAL. LHRH AND GALANIN PEPTIDES WERE DETECTED BY IMMUNOCYTOCHEMISTRY USING COLLOIDAL GOLD PARTICLES OF DIFFERENT SIZES. LHRH PEPTIDE IS MARKED BY SMALL, 5 nm GOLD PARTICLES, WHILE GALANIN IS TAGGED BY 15 nm GOLD PARTICLES. NOTE THE CO-EXPRESSION OF THE TWO DISTINCT PEPTIDES IN THE SAME SECRETORY GRANULE Basics of Neurobiology: Neurotransmitters II. 8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 9 www.itk.ppke.hu IN THE AXON TERMINAL, MOST GRANULES FORM A RESERVE POOL AND ONLY A FEW CAN BE SEEN PRIMED AND DOCKED AT THE PRESYNAPTIC MEMBRANE THE NEUROPEPTIDE RELEASE IS EXECUTED BY EXOCYTOSIS. THE PROCESSREQUIRES ATP AND CALCIUM. REACHING THE CRITICAL INTRACELLULAR CONCENTRATION OF CALCIUM NEEDS BURSTS OF ACTION POTENTIALS IN THE SYNAPTIC CLEFT, NEUROPEPTIDES BIND TO THEIR RECEPTORS THAT ARE COUPLED TO G PROTEINS IN PEPTIDERGIC SYNAPSES, RECYCLING OF NEUROPEPTIDES DOES NOT OCCUR. THEY RATHER DIFFUSE AND GET CLEAVED BY ENDOPEPTIDASES THE NEUTRAL ENDOPEPTIDASE, A METALLOENDOPEPTIDASE, LOCATED AT THE CELL SURFACE HYDROLYSES THE RELEASED PEPTIDES AT THE AMINO SIDE OF A HYDROPHOBIC RESIDUE THE ACTIVITY OF ENDOPEPTIDASES CAN LOCALLY REGULATE THE PEPTIDE CONCENTRATION AND CONSEQUENTLY, THE ACTIVATION OF THE RECEPTORS PROCESSING OF NEUROPEPTIDES Basics of Neurobiology: Neurotransmitters II. 8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 10 www.itk.ppke.hu NEUROPEPTIDE FAMILIES FAMILYNAME FAMILY MEMBERS TACHYKININS SUBSTANCE P,NEUROKININ A, NEUROKININ B OPIODPEPTIDES ENKEPHALIN, DYNORPHIN, ENDORPHIN PANCREATIC POLYPEPTIDE-RELATED NEUROPEPTIDEY, PEPTIDE YY, PANCREATIC POLYPEPTIDE INSULIN FAMILY INSULIN, INSULIN LIKE GROWTH FACTORI-II GASTRINS GASTRIN,CHOLECYSTOKININ SECRETINS VASOACTIVE INTESTINAL PEPTIDE (VIP), SECRETIN, GLUCAGON NEUROHYPOPHYSEAL HORMONES OXYTOCIN, VASOPRESSIN,NEUROPHYSIN I-II RELEASE-AND RELEASE-INHIBITING LHRH,TRH, CRH, GHRH, SOMATOSTATIN FEEDING PEPTIDES NEUROPEPTIDEY, AGOUTI RELATED PEPTIDE, ALPHA-MELANOCYTE STIMULATING HORMONE, NESFATIN, OREXIN Basics of Neurobiology: Neurotransmitters II. 8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 11 www.itk.ppke.hu SECRETED HORMONE ABBREVIATION PRODUCED BY EFFECT CORTICOTROPIN-RELEASING HORMONE CRH or CRF Parvocellular neurosecretory neurons Stimulate adrenocorticotropic hormone (ACTH) release from anterior pituitary THYROTROPIN-RELEASING HORMONE TRH, TRF Parvocellular neurosecretory neurons Stimulate thyroid-stimulating hormone (TSH) release from anterior pituitary Stimulate prolactin release from anterior pituitary GONADOTROPINRELEASING HORMONE GnRH or LHRH Parvocellular neurosecretory neurons Stimulate follicle-stimulating hormone (FSH) release from anterior pituitaryStimulate luteinizing hormone (LH) release from anterior pituitary DOPAMINE(PROLACTIN-INHIBITING HORMONE) DA or PIH Dopamine neurons of the arcuate nucleus Inhibitprolactin release from anterior pituitary GROWTH HORMONE-RELEASING HORMONE GHRH Parvocellular neurosecretory neurons Stimulate growth hormone (GH) release from anterior pituitary SOMATOSTATIN(GROWTH HORMONE-INHIBITING HORMONE) SS, GHIH, or SRIF Parvocellular neurosecretory neurons Inhibit Growth hormone (GH) release from anterior pituitaryInhibit thyroid-stimulating hormone (TSH) release from anterior pituitary OXYTOCIN Magnocellular neurosecretory cells Uterine contractionLactation (letdown reflex) VASOPRESSIN(ANTIDIURETIC HORMONE) ADH or AVP Magnocellular neurosecretory neurons Increases water permeability in the distal convoluted tubule and collecting duct of nephrons, thus promoting water reabsorption and increasing blood volume DOPAMINE(PROLACTIN-INHIBITING HORMONE) DA or PIH Dopamine neurons of the arcuate nucleus Inhibitprolactin release from anterior pituitary HYPOTHALAMIC HORMONES AND THEIR FUNCTIONS Basics of Neurobiology: Neurotransmitters II. 8/5/2011. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 12 www.itk.ppke.hu OPIOID PEPTIDES, THEIR ANALOGUES AND RECEPTORS NAME AMINO ACID SEQUENCE Phe-Gly-Gly-Phe-Thr-Gly-Ala-Arg-Lys-Ser-Ala-Arg-Lys-Leu-Ala-Asn-Gln Dynorphin A (1-17) Tyr-Gly-Gly-Phe-Leu-Arg-Arg-Ile-Arg-Pro-Lys-Leu-Lys-Trp-Asp-Asn-Gln Endomorphins 1 and 2 Tyr-Pro-Trp-Phe; Tyr-Pro-Phe-Phe Leu-and Met-enkephalin Nociceptin Endogenous opioid ß-Endorphin Enkephalins Dynorphins Nociceptin Synthetic agonist Morphine DSTBULET U50488H Codeine DPDPE Pentazocine Fentanyl Oxycodone? Pethidine Antagonists Naloxone Naloxone Naloxone Not naloxone ß-FNA Naltrindole nor-BNI Effector mechanism G protein G protein G protein G protein opens K+ opens K+ closes Ca2+ opens K+ channel channel channel channel Effects Hyperpolarization of neurons, inhibition of neurotransmitter release