Receptorarchitecture and Neural Systems...Receptorarchitecture and Neural Systems . Karl Zilles . Institute of Neuroscience and Medicine INM-1 . Research Centre Jülich . and . University

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

m

Transmitter Systems

Glutamate (ubiquitous; excitatory) GABA (ubiquitous; inhibitory) 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)

medial forebrain bundle

NMDA Receptor: Transmitter Glutamate/Effect: Excitation

NMDA receptors are highly permeant for Ca2+, and the ion channel is blocked in a voltage- and use-dependent manner by physiological concentrations of Mg2+ ions. These properties make them ideally suited for Hebbian processes in synaptic plasticity such as learning.

Metabotropic Glutamate Receptors: Transmitter Glutamate/Effect: Excitation

Glutamate

Adenosin A1 D1, D2, D4

D1, D2, D4

Dopamine

AMPA, NMDA, kainate

AMPA, NMDA, kainate

AMPA, NMDA, kainate

AMPA, NMDA, kainate

Glutamate

GABAA, bz.binding site




GABAB

GABAB


GABAB

GABA

nicotinic, M1, M2, M3

nicotinic, M1, M2, M3

Acetylcholine

α1, α2

α1, α2

Noradrenaline

5-HT1A, 5-HT2

5-HT1A, 5-HT2

Serotonin

Ionotropic glutamate AMPA receptor

Ionotropic glutamate NMDA receptor

Ionotropic glutamate Kainate receptor

Metabotropic glutamate mGluR 2/3 receptor

Metabotropic acetylcholine M2 receptor

Ionotropic glutamate GABAA receptor

• presynaptic • postsynaptic • autoreceptor • heteroreceptor

glutamate

acetylcholine

GABA

axon

axon

axon

dendrite

Inhibitory interneuron

Modulatory projection

neuron

Presynaptic excitatory pojection neuron

Postsynaptic Neuron

inhib/excit

How to map receptor distributions:

• in vivo receptor PET

• in vitro quantitative receptor autoradiography

• immunohistochemistry of receptor proteins and subunits

• transcriptomics

HG05/00

Sulcus centralis

slab 2 native

slab 2 frozen

at -70° C

slab 2

Quantitative in vitro Receptor Autoradiography: Method (1)

Gyrus praecentralis

Serial sections (20 µm) mounted onto glass slides

• Preincubation: re-hydrate sections. Remove endogenous substances

• Main incubation: buffer solution with [3H]-ligands which specifically bind to a given receptor type

• Washing step: stop binding procedure. Eliminate surplus [3H]-ligand and buffer salts

Binding protocol

Image processing

0

50

100

150

200

250

0 150000 350000

Radioactivity concentration (cpm)

Gre

y va

lues

Linearised and color-coded autoradiograph

grey values or colors encode

receptor densities

5-HT1A

LLK

SC D

a

+⋅=

1 [fmol/mg protein]

50

400

Regional Specificity (example: Amygdala)

and

Connectivity (example: Hippocampus)

muscarinic receptor M2 GABAA receptor NMDA receptor

basomedial basolateral

lateral

Amygdala

Receptor autoradiography

Structural MRI at 4 Tesla: spatial resolution 350 µm isotrop

bm bl l

sub

Kainate NMDA

AMPA

CA1 mol

CA1

mol CA3 CA2

Glutamatergic terminals of the perforant path and the Schaffer collaterals

Glutamatergic terminals of the mossy fibers

Glutamatergic terminals of the perforant path and the Schaffer collaterals

295 464 633 802

221 357 493 629 417 632 486 1061 fmol/mg protein fmol/mg protein

fmol/mg protein

mol

pp

CA3

CA2

CA1

Schaffer collaterals

mf

Genetic cell typing by Weissman & Lichtman (2006)

Principal Cortical Organization

Example: Primary Sensory Cortices

Cholinergic muscarinic M2 receptor [3H] oxotremorine-M Myelin staining

184 fmol/mg protein

262 341 420

Matching receptor- and myeloarchitecture

* *

* V1

V1

V1

layers II-IVa

layer IVc

Gennari‘s stripe V2

V2

V1

V1

Myelin

vertical meridian

vertical meridian

GABAA Receptor

V2

V2

V1

V1

vertical meridian

Sulcus calcarinus

Zilles, K., Palomero-Gallagher, N. , Schleicher, A.: Transmitter receptors and functional anatomy of the cerebral cortex. J. Anat. 205: 417-432 (2004)

vertical meridian

Human primary visual cortex V1

Macaque Brain

Human Brain

primary somatosensory cortex

primary auditory cortex

motor cortex

motor cortex sc

sc

Cholinergic muscarinic M2 receptor

primary auditory cortex

primary somatosensory cortex

184 fmol/mg protein 420

Multi-Receptor Fingerprints

Dopamine

Acetylcholine

Serotonin

Glutamate

Noradrenaline GABA

Low density

High density

AMPA Kainate NMDA M1 M2 M3 nicotinic

α1 α2 GABAA 5-HT1A 5-HT2 D1 D2

Multiple (multimodal) receptor organization in the human cerebral cortex

Zilles, K., Schleicher, A., Palomero-Gallagher, N., Amunts, K.: Quantitative analysis of cyto- and receptorarchitecture of the human brain, pp. 573-602. In: Brain Mapping: The Methods, 2nd edition (A.W. Toga and J.C. Mazziotta, eds.). Academic Press (2002)

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)

α1 D1 5-HT2

* *

* *

*

V1

V2

low high

*

*

*

*

GABAA GABAB nic α4β2 M1

mGluR2/3 kainate NMDA AMPA V2

*

Human primary visual cortex

5-HT1A

A receptor fingerprint is… AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

nACh

α1

α2

5-HT1A

5-HT2

D1

area 4

caudate putamen

0

500

1000

1500

2000

2500

3000

3b

0

1000

2000

3000 AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

N

α1

α2

5-HT1A

5-HT2

D1

4

0

1000

2000

3000 AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

N

α1

α2

5-HT1A

5-HT2

D1

V1

0

1000

2000

3000 AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

N

α1

α2

5-HT1A

5-HT2

D1

0

1000

2000

3000 AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

N

α1

α2

5-HT1A

5-HT2

D1

0

1000

2000

3000 AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

N

α1

α2

5-HT1A

5-HT2

D1

41

0

1000

2000

3000 AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

N

α1

α2

5-HT1A

5-HT2

D1

Fingerprints: modality specificity primary somatosensory primary motor primary auditory

V2 V3

primary (V1), and higher (V2, V3) visual areas

Ventral visual stream in human extrastriate cortex (pFus)

(pFus)

FG1 FG2 (pFus) GABAB

GABAA bz b.s.

NMDA

5-HT1A

Receptor fingerprint of FG1

Receptor fingerprint of FG2

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: 205-220 (2015)

ventral stream

Hierarchical Cluster Analysis

auditory

somatosensory

motor

On the way to understand mechanisms behind receptor fingerprints: Conditional 5-HT1A receptor knockout

WT

KO

64 636 fmol/mg protein

5-HT1A receptor

109 1091 fmol/mg protein

α1 receptor

What receptors tell us about laminar segregation and input-output relations

in the cerebral cortex

molecular layer I

outer granular layer II

inner granular layer IV

outer pyramidal layer III

inner pyramidal layer V

polymorphic layer VI

CYTOARCHITECTURE

thalamo- cortical

input

cortico- cortical

input

CONNECTIVITY SYNAPTIC DENSITY

cortico- -cortical, -striatal,

-thalamic, -bulbar, and

-spinal output

0

1000

2000

3000 AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

N

α1

α2

5-HT1A

5-HT2

D1

L. I

0

1000

2000

3000 AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

N

α1

α2

5-HT1A

5-HT2

D1 Ll. II-III

Receptor Fingerprints of the primary visual cortex: layer specificity

0

1000

2000

3000 AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

N

α1

α2

5-HT1A

5-HT2

D1

L. IV

L. V

0

1000

2000

3000 AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

N

α1

α2

5-HT1A

5-HT2

D1 L. VI

0

1000

2000

3000 AMPA

kainate

NMDA

GABAA

GABAB

BZ

M1

M2 M3

N

α1

α2

5-HT1A

5-HT2

D1 I II

IV

III

I II

III

V

VI

I II

IV

III

V

VI

In/output Synapse

Receptor Fingerprints

and

Complex Neural Systems

7

47

IFS1/IFJ 9

45p 44v

46 PFop

PFt PFcm

PFm

PF

pSTG/STS

4d

44d

3b

Te1

Te2 45a

PGa PGp

4v

V1

FG1 FG2

32

Sentence comprehension-related and non-related brain regions

15 transmitter receptor types and cognitive systems

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

fMRI defined regions “sentence comprehension task”

AM

PA

Kai

nate

N

MD

A

GA

BA A

G

AB

AB

B

Z

750

100

850

75

1700

100

2200

100

3000

100

3500

100

V1 I III IVa IVb IVc V VI

PFm 44d I II

IIIab IIIc IV V

VI

47 4d pSTG / STS

IFS1 / IFJ 45

I II IIIab IIIc IV V

VI

I II

IIIc IV V

VI

I II

IIIc IV V

VI

I II

IIIc

V

VI

I II

IIIc IV V

VI

I II

IIIc IV V

VI

IIIab IIIab IIIab IIIab IIIab II

PFm

47

IFS1/IFJ

45 pSTG/STS

44d

4d

V1

Laminar distribution of

multiple receptors in different

cortical areas


multimodal association

primary sensory

language network

Euclidean Distance 0.5 1 1.5 2 2.5 3

FG1 FG2

7 9

46 32

44v 44d

47 45a

pSTG/STS Te2 45p

IFS1/IFJ 4v 4d

3b Te1

V1

PF PFm PGa

PFcm PFop

PFt PGp

Left hemisphere (language dominant side)


Hierarchical Cluster Analysis of Multi-receptor Fingerprints Reveals Principles of Functional Organization


Euclidean Distance

0.5 1 1.5 2 2.5 3

FG2 FG1 PGp Te2 PFt

PFop PFcm

PGa 7

PFm PF

pSTG/STS IFS1/IFJ

45p 45a 44v 44d

32 9

46 47 4d 4v V1

Te1 3b

Right hemisphere


primary sensory

Broca‘s region


Hierarchical Cluster Analysis of Multi-receptor Fingerprints Reveals Principles of Functional Organization

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

Thanks to:

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Receptorarchitecture and Neural Systems...Receptorarchitecture and Neural Systems . Karl Zilles . Institute of Neuroscience and Medicine INM-1 . Research Centre Jülich . and . University