Receptorarchitecture and Neural Systems
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1 Receptorarchitecture and Neural Systems Karl Zilles Institute of Neuroscience and Medicine INM-1 Research Centre Jülich and University Hospital of Psychiatry, Psychotherapy and Psychosomatics RWTH Universität Aachen
2 Transmitter Systems Glutamate GABA (ubiquitous; excitatory) (ubiquitous; inhibitory) medial forebrain bundle m Acetylcholine (basal forebrain Ch1 Septum Ch2 vertical band Ch3 horizontal b. Ch4 NbM) Dopamine (substantia nigra, ventral tegmental area VTA) Noradrenaline (locus coeruleus) Serotonin (raphe nuclei)
3 Receptors are protein complexes in the cell membrane, to which transmitters selectively bind. ionotropic receptors have an integrated ion channel and regulate ion fluxes between extra- and intracellular space metabotropic receptors are linked to intracellular second messenger systems, and control metabolic pathways, ion channels, gene activity, etc.
4 Example: Ionotropic Transmitter Receptors presynaptic membrane postsynaptic membrane postsynaptic ionotropic receptor R ion channel axonal bouton transmitter presynaptic ionotropic receptor
5 5-HT 1A, 5-HT 2 AMPA, NMDA, kainate GABA B Adenosin A1 D1, D2, D4 a 1, a 2 AMPA, NMDA, kainate AMPA, NMDA, kainate GABA A, bz.binding site nicotinic, M 1, M 2, M 3 a 1, a 2 GABA A, bz.binding site nicotinic, M 1, M 2, M 3 D1, D2, D4 GABA B 5-HT 1A, 5-HT 2 GABA A, bz.binding site GABA B AMPA, NMDA, kainate GABA A, bz.binding site GABA A, bz.binding site Glutamate GABA Acetylcholine Dopamine Noradrenaline Serotonin
6 How to map receptor distributions: in vivo receptor PET in vitro quantitative receptor autoradiography immunohistochemistry of receptor proteins and subunits transcriptomics
7 Quantitative in vitro Receptor Autoradiography: Method (1) Sulcus centralis Gyrus praecentralis HG05/00 slab 2 slab 2 native slab 2 frozen at -70 C
8 Serial sections (20 µm) mounted onto glass slides
9 Binding protocol Preincubation: re-hydrate sections. Remove endogenous substances Main incubation: buffer solution with [ 3 H]-ligands which specifically bind to a given receptor type Washing step: stop binding procedure. Eliminate surplus [ 3 H]-ligand and buffer salts
![binding [ 3 H]-ligand (nm) + displacer](/docs-images/95/124549540/images/10-1.jpg "(µm) Specific binding = total binding")
10 Total and nonspecific binding Total binding [ 3 H]-ligand (nm) Nonspecific binding [ 3 H]-ligand (nm) + displacer (µm) Specific binding = total binding nonspecific binding
11 Grey values Image processing 5-HT 1A Receptor density (fmol/mg protein) Linearised autoradiograph grey values code receptor densities
12 M 2 receptor [ 3 H] oxotremorine-m Myelin staining fmol/mg protein * * Primary visual cortex
13 V2 V2 V1 Sulcus calcarinus vertical vertical meridian meridian V1 V1 V1 Gennari s stripe V2 V2 vertical meridian Myelin GABA A Receptor Zilles, K., Palomero-Gallagher, N., Schleicher, A.: Transmitter receptors and functional anatomy of the cerebral cortex. J. Anat. 205: (2004)
14 Entorhinal-Hippocampal System
15 fmol/mg protein Schaffer collateral CA2 CA3 fimbria CA2 CA3 mol AMPA mossy fibres CA1 CA1 mol perforant path sub Glutamatergic terminals of the perforant path and the Schaffer collaterals fmol/mg protein fmol/mg protein NMDA Kainate CA1 mol Glutamatergic terminals of the perforant path and the Schaffer collaterals Glutamatergic terminals of the mossy fibers
16 Primary Sensory Cortices
17 Cholinergic muscarinic M2 receptor low density high density sc motor cortex primary somatosensory cortex motor cortex sc primary somatosensory cortex primary auditory cortex primary auditory cortex Human Brain Macaque Brain
18 Receptor Fingerprints
19 Low denisty Multimodal visualization of receptor organization in human cerebral cortex AMPA Kainate NMDA M1 M2 M3 nicotinic High density Glutamate Acetylcholine a 1 a 2 GABA A 5-HT 1A 5-HT 2 D1 D2 Noradrenaline GABA Serotonin Dopamine Zilles, K., Schleicher, A., Palomero-Gallagher, N., Amunts, K.: Quantitative analysis of cyto- and receptorarchitecture of the human brain, pp In: Brain Mapping: The Methods, 2nd edition (A.W. Toga and J.C. Mazziotta, eds.). Academic Press (2002)
20 A receptor fingerprint is 5-HT 2 D 1 AMPA kainate NMDA 5-HT 1A a GABA A GABA B a 1 BZ caudate nach M 3 M 2 M 1 putamen area 4
21 Receptor Fingerprints D 1 AMPA 3000 kainate D 1 AMPA 3000 kainate 5-HT NMDA 5-HT NMDA 5-HT 1A 1000 GABA A 5-HT 1A 1000 GABA A a 2 0 GABA B a 2 0 GABA B a 1 BZ a 1 BZ N M 1 N M 1 M 3 M 2 M 3 M 2 primary motor cortex Hippocampus CA1-3
22 What receptors tell us about laminar segregation and input-output relations in the cerebral cortex
23 CYTOARCHITECTURE CONNECTIVITY SYNAPTIC DENSITY molecular layer I outer granular layer II outer pyramidal layer III thalamocortical input corticocortical input inner granular layer IV inner pyramidal layer polymorphic layer V VI cortico- -cortical, -striatal, -thalamic, -bulbar, and -spinal output
24 BZ NMDA Kainate AMPA 4d IFS1/IFJ PFm 44d I II IIIab IIIc IV V VI 45 I II IIIab IIIc IV V VI IFS1 / IFJ pstg / STS V1 4d 47 PFm I I I II II II I I I II III II II IIIab IIIab IIIab IIIab IIIab IIIc IVa IIIc IIIc IIIc IV IV IVb IIIc IV IV V V IVc V V V V VI VI VI VI VI VI d 45 pstg/sts V1 GABA A GABA B
25 Receptor Fingerprints of the primary visual cortex: layer specificity L. I 5-HT 2 D 1 AMPA kainate NMDA Ll. II-III 5-HT 2 D 1 AMPA kainate NMDA L. IV 5-HT 2 D 1 AMPA kainate NMDA 5-HT 1A 1000 GABA A 5-HT 1A 1000 GABA A 5-HT 1A 1000 GABA A a 2 GABA B a 2 GABA B a 2 GABA B a 1 BZ a 1 BZ a 1 BZ N M 1 N M 1 N M 1 M 3 M 2 M 3 M 2 M 3 M 2 L. V AMPA D kainate L. VI D 1 5-HT NMDA 5-HT 2 AMPA kainate NMDA 5-HT 1A 1000 GABA A 5-HT 1A 1000 GABA A a 2 0 GABA B a 2 0 GABA B a 1 BZ a 1 BZ N M 1 N M 1 M 3 M 2 M 3 M 2
26 15 transmitter receptor types and cognitive systems Sentence comprehension-related and non-related brain regions 4d 7 4v 3b PFm 9 IFS1/IFJ PFt PF PGa 44d PFcm PGp 46 45p Te1 PFop 45a 44v Te2 pstg/sts 47 V1 FG1 FG2 32 fmri defined regions sentence comprehension task K. Zilles, M. Bacha-Trams, N. Palomero-Gallagher, K. Amunts, A.D. Friederici (2015). Common molecular basis of the sentence comprehension network revealed by neurotransmitter receptor fingerprints. Cortex 63: 79-89
27 Left hemisphere (language dominant side) Right hemisphere multimodal association primary sensory language network multimodal association PGa PFm PF PGp PFt PFop PFcm V1 3b Te1 4d 4v IFS1/IFJ 45p Te2 pstg/sts 45a 47 44d 44v FG2 FG1 3b Te1 V1 4v 4d d 44v 45a 45p IFS1/IFJ pstg/sts PF PFm 7 PGa PFcm PFop PFt Te2 PGp FG1 FG Euclidean Distance Euclidean Distance K. Zilles, M. Bacha-Trams, N. Palomero-Gallagher, K. Amunts, A.D. Friederici (2015). Common molecular basis of the sentence comprehension network revealed by neurotransmitter receptor fingerprints. Cortex 63: 79-89
28 Euclidean distance (Ward linkage) 24a 24b Cluster analysis of the human cingulate cortex: A multi-region model based on receptor fingerprints 24cv 24cd 32 autonomic control a24a' a24b' 32' 24c'd 24c'v p24a' p24b' 24dv 24dd 23d 23c 31 d23 v m 29l Palomero-Gallagher, N., Vogt, B.A., Schleicher, A., Mayberg, H.S., Zilles, K. (2009) Receptor architecture of human cingulate cortex: Evaluation of the four-region neurobiological model. Human Brain Mapping 30: emotion ACC decision amcc MCC motor control pmcc visuospatial attention memory PCC limbic memory RSC 0.5 0
29 Thanks to: Institute of Neuroscience and Medicine (INM-1), Research Centre Jülich Katrin Amunts Mareike Bacha-Trams Svenja Caspers Nicola Palomero-Gallagher Axel Schleicher
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