2011.10.14.. 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 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 2 Peter Pazmany Catholic University Faculty of Information Technology BEVEZETÉS A FUNKCIONÁLIS NEUROBIOLÓGIÁBA INTRODUCTION TO FUNCTIONAL NEUROBIOLOGY www.itk.ppke.hu By Imre Kalló Contributed by: Tamás Freund, Zsolt Liposits, Zoltán Nusser, László Acsády, Szabolcs Káli, József Haller, Zsófia Maglóczky, Nórbert Hájos, Emilia Madarász, György Karmos, Miklós Palkovits, Anita Kamondi, Lóránd Erőss, Róbert Gábriel, Zoltán Kisvárday, Zoltán Vidnyánszky Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 3 www.itk.ppke.hu Electrophysiology_II.Imre Kalló & Norbert HájosPázmány Péter Catholic University, Faculty of Information Technology I. In vitro and in vivo recording techniques. II. Firing pattern of different cell types. III. Extracts from the studies on intercellular communications (studies on gap junctions, dendritic action potentials, synaptic currents, noise-analysis, saturation of receptors by neurotransmitters, quantal analysis, paired recording of neurons, combination of electrophysiology with imaging techniques) Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 4 www.itk.ppke.hu In vivo recordings Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 5 www.itk.ppke.hu Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 6 www.itk.ppke.hu Ion concentrations and Equilibrium Potentials Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 7 www.itk.ppke.hu In vitro electrophysiological recording Whole-cell Recordings Stimulation Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 8 www.itk.ppke.hu Equipment used chamber submergedstage2 Interface recording chamber Submerged recording chamber Introduction to functional neurobiology: Electrophysiology gapjunction 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 9 www.itk.ppke.hu Electric synapses –gap junction Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 10 www.itk.ppke.hu Electric synapses –gap junction 100ms 5mV 1.5mV 1 2 2.8% Ref:SzabadicsJ,LorinczA,TamásG.Betaandgammafrequencysynchronizationbydendriticgabaergicsynapsesandgapjunctionsinanetworkofcorticalinterneurons.JNeurosci.2001Aug1;21(15):5824-31. Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 11 www.itk.ppke.hu Active dendrites –in vitro recording Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 12 www.itk.ppke.hu Active dendrites –in vitro recording Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 13 www.itk.ppke.hu Synaptic connection Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 14 www.itk.ppke.hu EPSP Excitatory neurotransmission (e.g. glutamate or acetylcholin receptors) Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 15 www.itk.ppke.hu Synaptic transmission at the neuromuscular junction FFigu Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 16 www.itk.ppke.hu Noise-analysis of synaptic currents fffig1 .2= i* Im-Im2/ N .2-variance i–single channel current Im–mean current N –number of channel, which generate current Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 17 www.itk.ppke.hu Studies on synaptic transmission I. SynTransm1.jpg SynTransm2.jpg Electric stimulation experiments Ref:HájosN,FreundTF.Distinctcannabinoidsensitivereceptorsregulatehippocampalexcitationandinhibition.ChemPhysLipids.2002Dec31;121(1-2):73-82.Review. Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 18 www.itk.ppke.hu Studies on synaptic transmission I. SynTransm4.jpg SynTransm5.jpg Minimalstimulation experiments Ref:HájosN,KatonaI,NaiemSS,MacKieK,LedentC,ModyI,FreundTF.CannabinoidsinhibithippocampalGABAergictransmissionandnetworkoscillations.EurJNeurosci.2000Sep;12(9):3239-49. Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 19 www.itk.ppke.hu Studies on synaptic transmission II. SynTransm8.jpg SynTransm7.jpg Action potential-dependent synaptic transmission Ref:KatonaI,RanczEA,AcsadyL,LedentC,MackieK,HajosN,FreundTF.DistributionofCB1cannabinoidreceptorsintheamygdalaandtheirroleinthecontrolofGABAergictransmission.JNeurosci.2001Dec1;21(23):9506-18. Introduction to functional neurobiology: Electrophysiology SynTransm10.jpg 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 20 www.itk.ppke.hu Studies on synaptic transmission II. SynTransm9.jpg Action potential-independent synaptic transmission Ref:KatonaI,RanczEA,AcsadyL,LedentC,MackieK,HajosN,FreundTF.DistributionofCB1cannabinoidreceptorsintheamygdalaandtheirroleinthecontrolofGABAergictransmission.JNeurosci.2001Dec1;21(23):9506-18. Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 21 www.itk.ppke.hu Ultrastructure of axon terminals fig4 Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 22 www.itk.ppke.hu Saturation of synaptic neurotransmitter-receptors Zolpidem-egybenzodiazepine származékhatása Ref:HájosN,NusserZ,RanczEA,FreundTF,ModyI.Celltype-andsynapse-specificvariabilityinsynapticGABAAreceptoroccupancy.EurJ Neurosci.2000Mar;12(3):810-8. Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 23 www.itk.ppke.hu Large variety of neurotransmitter receptors: the GABAa receptors as an example PostsynapticR1.jpg PostsynapticR2.jpg Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 24 www.itk.ppke.hu Large variety of neurotransmitter receptors: the GABAa receptors as an example GABAaR1.jpg GABAaR2.jpg Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 25 www.itk.ppke.hu Ultrastructure of axon terminals fig4 Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 26 www.itk.ppke.hu The release of transmitters is quantal FFige Introduction to functional neurobiology: Electrophysiology Graph2.jpg Graph1.jpg 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 27 www.itk.ppke.hu Control Kindled Frequency pA Frequency pA Frequency X0= 0.6±6.2 X2=75.7±11.6 X4=155.9±7.9 X3=116.0±13.2 X1= 38.9±9.8 q AV= 38.9 pA Amplitude (pA) X0= -0.13±8.3 X2=107.3±18.7 X4=217.9±19.4 X3=166.6±17.8 X1= 56.4±15.2 q AV= 54.5 pA Amplitude (pA) 40 pA 10 ms Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 28 www.itk.ppke.hu Kindled Control Ref:Nusser Z, Hájos N, SomogyiP, ModyI. Increased number of synaptic GABA(A) receptors underlies potentiationat hippocampalinhibitory synapses. Nature. 1998Sep 10;395(6698): 172-7. Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 29 www.itk.ppke.hu Kindled Control Ref:Nusser Z, Hájos N, SomogyiP, ModyI. Increased number of synaptic GABA(A)receptors underlies potentiationat hippocampalinhibitory synapses. Nature. 1998Sep 10;395(6698):172-7. Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 30 www.itk.ppke.hu Paired recording of neurons green cell.tif 00000A8DData B3A11CF3: infrared cell.tif 00000A8DData B3A11CF3: patched cells.tif 00000A8DData B3A11CF3: red cell.tif 00000A8DData B3A11CF3: blue cell.tif 00000A8DData B3A11CF3: bicoloc.tiff 00000A8DData B3A11CF3: 4 coloc.tif 00000A8DData B3A11CF3: EGFP, Biocytinand CR colocalization Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 31 www.itk.ppke.hu Paired recording of neurons Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 32 www.itk.ppke.hu Paired recording of neurons (dual or triple recording) P B M The quality of neurotransmission is determined by the type of postsynaptic cell!!! Bistratifiedcell (dendritic inhibitory cell) Multipolar cell (basket cell) Pyramidal cell Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 33 www.itk.ppke.hu Difference of Ca2+ tranzients evoked by action potential is large in the axon terminals of pyramidal cells Ref:Koester HJ, SakmannB. Calcium dynamics associated with action potentials iningle nerve terminals of pyramidal cells in layer 2/3 of the young ratneocortex. J Physiol. 2000 Dec 15;529 Pt 3:625-46. Introduction to functional neurobiology: Electrophysiology 2011.10.14. TÁMOP –4.1.2-08/2/A/KMR-2009-0006 34 www.itk.ppke.hu Tracing of gamma oscillation in the CA3 region with voltage-sensitive fluorescent dyes - CSD VSD pyr pyr pyr rad rad rad dep hyp time Ref:Mann et al Neuron. 2005 Jan 6;45(1):105-17.