Receptorarchitecture and Neural Systems

Transcription

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 NMDA Receptor: Transmitter Glutamate/Effect: Excitation NMDA receptors are highly permeant for Ca 2+, and the ion channel is blocked in a voltage- and use-dependent manner by physiological concentrations of Mg 2+ ions. These properties make them ideally suited for Hebbian processes in synaptic plasticity such as learning.

4 Metabotropic Glutamate Receptors: Transmitter Glutamate/Effect: Excitation Glutamate

5 5-HT 1A, 5-HT 2 AMPA, NMDA, kainate GABA B Adenosin A1 D1, D2, D4 α 1, α 2 AMPA, NMDA, kainate AMPA, NMDA, kainate GABA A, bz.binding site nicotinic, M 1, M 2, M 3 α 1, α 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 Modulatory projection neuron axon presynaptic postsynaptic autoreceptor heteroreceptor glutamate acetylcholine GABA Presynaptic excitatory pojection neuron axon dendrite Postsynaptic Neuron inhib/excit Inhibitory interneuron axon Ionotropic glutamate AMPA receptor Ionotropic glutamate NMDA receptor Ionotropic glutamate Kainate receptor Metabotropic glutamate mglur 2/3 receptor Metabotropic acetylcholine M 2 receptor Ionotropic glutamate GABA A receptor

7 How to map receptor distributions: in vivo receptor PET in vitro quantitative receptor autoradiography immunohistochemistry of receptor proteins and subunits transcriptomics

8 Quantitative in vitro Receptor Autoradiography: Method (1) Sulcus centralis Gyrus praecentralis HG05/00 slab 2 slab 2 native slab 2 frozen at -70 C

9 Serial sections (20 µm) mounted onto glass slides

10 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

11 Image processing K D + L C = 1 [fmol/mg protein] S L a HT 1A Grey values Radioactivity concentration (cpm) 50 Linearised and color-coded autoradiograph grey values or colors encode receptor densities

12 Regional Specificity (example: Amygdala) and Connectivity (example: Hippocampus)

13 Amygdala l bm bl basomedial basolateral lateral Structural MRI at 4 Tesla: spatial resolution 350 µm isotrop GABA NMDA muscarinic receptor M A receptor 2 Receptor autoradiography

14 CA3 mf 802 CA2 633 CA fmol/mg protein AMPA mol mol Schaffer collaterals CA1 pp CA1 CA2 Genetic cell typing by Weissman & Lichtman (2006) 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

15 Principal Cortical Organization Example: Primary Sensory Cortices

16 Matching receptor- and myeloarchitecture Cholinergic muscarinic M 2 receptor [ 3 H] oxotremorine-m Myelin staining fmol/mg protein layers II-IVa V1 layer IVc * V1 V1 * *

17 Human primary visual cortex V1 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)

18 Cholinergic muscarinic M2 receptor fmol/mg protein sc motor cortex primary somatosensory cortex motor cortex sc primary somatosensory cortex primary auditory cortex primary auditory cortex Human Brain Macaque Brain

19 Multi-Receptor Fingerprints

20 Low density Multiple (multimodal) receptor organization in the human cerebral cortex AMPA Kainate NMDA M1 M2 M3 nicotinic High density Glutamate Acetylcholine α 1 α 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)

21 V2 low AMPA Human primary visual cortex NMDA kainate mglur2/3 high V1 * * * V2 GABA A GABA B M 1 nic α 4 β 2 * * * α 1 5-HT 1A 5-HT 2 D 1 * * * * Zilles, K., Palomero Gallagher, N.: Comparative analysis of receptor subtypes that identify primary cortical sensory areas. In (Kaas, J., Striedter, G., Krubitzer, L., Herculano-Houzel, S., Preuss, T., eds.) Evolution of the Nervous System. Elsevier, Oxford (in press)

22 A receptor fingerprint is 5-HT 2 D AMPA kainate NMDA 5-HT 1A α GABA A GABA B α 1 BZ caudate nach M 3 M 2 M 1 putamen area 4

23 Fingerprints: modality specificity primary somatosensory primary motor primary auditory 3b 5-HT 1A 5-HT 2 D 1 α 2 AMPA kainate NMDA GABA A GABA B 4 5-HT 1A 5-HT 2 D 1 α 2 AMPA kainate NMDA GABA A GABA B 41 5-HT 1A 5-HT 2 D 1 α 2 AMPA kainate NMDA GABA A GABA B α 1 BZ α 1 BZ α 1 BZ N M 3 M 2 M 1 N M 3 M 2 M 1 N M 3 M 2 M 1 primary (V1), and higher (V2, V3) visual areas V1 5-HT 2 D 1 AMPA kainate NMDA V2 5-HT 2 D 1 AMPA kainate NMDA V3 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 α 2 0 GABA B α 2 0 GABA B α 2 0 GABA B α 1 BZ α 1 BZ α 1 BZ N M 1 N M 1 N M 1 M 3 M 2 M 3 M 2 M 3 M 2

24 Ventral visual stream in human extrastriate cortex (pfus) (pfus)

25 FG1 GABA NMDA B FG2 (pfus) GABA 5-HT A bz b.s. 1A

26 Receptor fingerprint of FG1 Hierarchical Cluster Analysis ventral stream Receptor fingerprint of FG2 auditory somatosensory motor Caspers, J., Palomero-Gallagher, N., Caspers, S., Schleicher, A., Amunts, K., Zilles, K.: Receptor architecture of cytoarchitectonic visual areas FG1 and FG2 of the posterior fusiform gyrus. Brain Struct. Funct. 220: (2015)

27 On the way to understand mechanisms behind receptor fingerprints: Conditional 5-HT 1A receptor knockout 5-HT 1A receptor α 1 receptor 64 fmol/mg protein fmol/mg protein 1091 WT KO

28 What receptors tell us about laminar segregation and input-output relations in the cerebral cortex

29 CYTOARCHITECTURE molecular layer I outer granular layer II outer pyramidal layer III inner granular layer IV inner pyramidal layer V polymorphic layer VI CONNECTIVITY thalamocortical input corticocortical input cortico-cortical, -striatal, -thalamic, -bulbar, and -spinal output SYNAPTIC DENSITY

30 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 α 2 0 GABA B α 2 0 GABA B α 2 0 GABA B α 1 BZ α 1 BZ α 1 BZ N M 1 N M 1 N M 1 M 3 M 2 M 3 M 2 M 3 M 2 In/output Synapse L. V AMPA D kainate L. VI D 1 5-HT NMDA 5-HT 2 AMPA kainate NMDA I II I II I II 5-HT 1A α 2 α 1 N M 1 GABA A BZ GABA B 5-HT 1A α 2 α 1 N M 1 GABA A BZ GABA B III IV V III III IV V M 3 M 2 M 3 M 2 VI VI

31 Receptor Fingerprints and Complex Neural Systems

32 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

33 Laminar distribution of multiple receptors in different cortical areas AMPA Kainate 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 75 IFS1/IFJ 44d 45 PFm pstg/sts NMDA BZ GABA B GABA A V

34 Hierarchical Cluster Analysis of Multi-receptor Fingerprints Reveals Principles of Functional Organization 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 Euclidean Distance Left hemisphere (language dominant side) 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

35 Hierarchical Cluster Analysis of Multi-receptor Fingerprints Reveals Principles of Functional Organization primary sensory multimodal association Broca s region multimodal association 3b Te1 V1 4v 4d d 44v 45a 45p IFS1/IFJ pstg/sts PF PFm 7 PGa PFcm PFop PFt Te2 PGp FG1 FG2 Euclidean Distance Right hemisphere 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

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37 Thanks to: Institute of Neuroscience and Medicine (INM-1), Research Centre Jülich: Katrin Amunts Mareike Bacha-Trams Julian Caspers Svenja Caspers Nicola Palomero-Gallagher Axel Schleicher Stanford University: Kalanit Grill-Spector Kevin Weiner MPI Leipzig: Angela Friederici UCLA, Ahmanson-Lovelace Brain Mapping Center: John C. Mazziotta, Arthur Toga