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.
BEVEZETÉS A FUNKCIONÁLIS NEUROBIOLÓGIÁBA
INTRODUCTION TO
FUNCTIONAL NEUROBIOLOGY
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
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 3
Electrophysiology_II.
Imre Kalló & Norbert Hájos
Pázmány 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)
In vivo recordings
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 5
Ion concentrations and Equilibrium Potentials
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 7
In vitro
electrophysiological recording
Recordings Stimulation
Equipment used
Interface recording chamber Submerged recording chamber
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 9
Electric synapses – gap junction
Electric synapses – gap junction
100ms 5mV 1.5mV
1 2
2.8%
Ref: Szabadics J, Lorincz A, Tamás G. Beta and gamma frequency synchronization by dendritic gabaergic synapses and gap junctions in a network of cortical interneurons. J Neurosci.
2001 Aug 1;21(15):5824-31.
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 11
Active dendrites – in vitro recording
Active dendrites – in vitro recording
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 13
Synaptic connection
Excitatory neurotransmission
(e.g. glutamate or acetylcholin receptors)
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 15
Noise-analysis of synaptic currents
σ2 = i * Im- Im2/ N
σ2 - variance
i – single channel current Im – mean current
N – number of channel, which generate current
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 17
Studies on synaptic transmission I.
Electric stimulation experiments
Ref: Hájos N, Freund TF. Distinct cannabinoid sensitive receptors regulate hippocampal excitation and inhibition. Chem Phys Lipids. 2002 Dec 31;121(1-2):73-82. Review.
Studies on synaptic transmission I.
Minimal stimulation experiments
Ref: Hájos N, Katona I, Naiem SS, MacKie K, Ledent C, Mody I, Freund TF. Cannabinoids inhibit hippocampal GABAergic transmission and network oscillations. Eur J Neurosci. 2000 Sep;12(9):3239-49.
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 19
Studies on synaptic transmission II.
Action potential-dependent synaptic transmission
Ref: Katona I, Rancz EA, Acsady L, Ledent C, Mackie K, Hajos N, Freund TF. Distribution of CB1 cannabinoid receptors in the amygdala and their role in the control of GABAergic transmission. J Neurosci. 2001 Dec 1;21(23):9506-18.
Studies on synaptic transmission II.
Action potential-independent synaptic transmission
Ref: Katona I, Rancz EA, Acsady L, Ledent C, Mackie K, Hajos N, Freund TF. Distribution of CB1 cannabinoid receptors in the amygdala and their role in the control of GABAergic transmission. J Neurosci. 2001 Dec 1;21(23):9506-18.
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 21
Ultrastructure of axon terminals
Saturation of synaptic neurotransmitter-receptors
Zolpidem - egy benzodiazepine származék hatása
Ref: Hájos N, Nusser Z, Rancz EA, Freund TF, Mody I. Cell type- and synapse-specific variability in synaptic GABAA receptor occupancy. Eur J
Neurosci. 2000 Mar;12(3):810-8.
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 23
the GABAa receptors as an example
Large variety of neurotransmitter receptors:
the GABAa receptors as an example
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 25
Ultrastructure of axon terminals
The release of transmitters is quantal
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 27
-10 -5 0 5 10
0 60 120 180
pA
-10 -5 0 5 10
0 50 100 150 200
pA
0 50 100 150 200 250 300 0
10 20 30
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)
0 50 100 150 200 250 300 0
5 10 15 20 25
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
Kindled
Control
Ref: Nusser Z, Hájos N, Somogyi P, Mody I.
Increased number of synaptic GABA(A) receptors underlies
potentiation at hippocampal inhibitory synapses. Nature.
1998 Sep 10;395(6698):
172-7.
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 29
Kindled
Control Ref: Nusser Z, Hájos N, Somogyi P, Mody I. Increased number of synaptic GABA(A) receptors underlies
potentiation at hippocampal inhibitory synapses. Nature. 1998 Sep
10;395(6698): 172-7.
Paired recording of neurons
EGFP, Biocytin and CR colocalization
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 31
Paired recording of neurons (dual or triple recording)
P B M
The quality of neurotransmission is determined by the type of postsynaptic cell!!!
Bistratified cell
(dendritic inhibitory cell) Multipolar cell (basket cell) Pyramidal
cell
2011.10.14. TÁMOP – 4.1.2-08/2/A/KMR-2009-0006 33
Difference of Ca
2+tranzients evoked by action potential is large in the axon terminals of pyramidal cells
Ref: Koester HJ, Sakmann B. Calcium dynamics associated with action potentials in ingle nerve terminals of pyramidal cells in layer 2/3 of the young rat neocortex. J Physiol. 2000 Dec 15;529 Pt 3:625-46.
Tracing of gamma oscillation in the CA3 region with voltage- sensitive fluorescent dyes
-
CSD
VSD pyr
pyr pyr rad
rad rad
dep hyp
Ref: Mann et al Neuron. 2005 Jan 6;45(1):105-17. time
