The complexity and simplicity of dopamine neurons

J. Paul Bolam

MRC Anatomical Neuropharmacology Unit

Oxford
The normality, complexity, simplicity and
subtlety of dopamine neurons
J. Paul Bolam

MRC Anatomical Neuropharmacology Unit

Oxford
The complexity and simplicity of dopamine neurons

Organisation of dopamine neurons at level of SNC

(normality)
Projections of dopamine neurons (complexity)
Responses of dopamine neurons (simplicity)
The effects of dopamine neurons (subtlety)
Dopamine neurons in SNC

DAT immunolabelling
SNC

SNR
TH immunolabelling
Dopamine neurons in SNC

Bentivoglio and Morelli (2005) Handbook Chem. Neuroanat 21:1-107

Single cell labelling
Afferents of dopamine neurons

GABAergic afferents

Striatum

Globus pallidus

Substantia nigra reticulata

Afferents of dopamine neurons
GABA

GABAergic afferents TH

Striatum

Globus pallidus

TH
Substantia nigra reticulata

GAD
Afferents of dopamine neurons

GABAergic afferents

Striatum

Globus pallidus

Substantia nigra reticulata

Fujiyama, Stephenson and Bolam (2002) Eur. J. Neurosci. 15:1961-1975

Afferents of DA
neurons GABA and GABAA receptors

GABAergic afferents

Striatum

Globus pallidus

SNR
 GABA and GABAA receptors
Afferents of DA
neurons

GABAergic afferents

Striatum

Globus pallidus

SNR
Afferents of dopamine neurons
GABAB receptors

GABAergic afferents

Striatum

Globus pallidus

Substantia nigra reticulata

Boyes and Bolam (2003) Eur. J. Neurosci. 18:3279-3293

Afferents of dopamine neurons

GABAergic afferents
GABAB receptors
Striatum

GP

SNR

Boyes and Bolam (2003) Eur. J. Neurosci. 18:3279-3293

Afferents of dopamine neurons

Glutamatergic afferents

Subthalamic nucleus

Pedunculopontine
nucleus

Cortex

Amygdala
Afferents of dopamine neurons

Glutamatergic afferents
VGLUT 2
STN

PPN

Cortex

Amygdala
Afferents of dopamine neurons
Glutamatergic afferents

STN

PPN

Cortex

Amygdala

Chatha, Bernard, Streit and Bolam, (2000) Neuroscience 101:1037-1051.

Afferents of dopamine neurons

Glutamatergic afferents

STN

PPN

Cortex

Amygdala

Chatha, Bernard, Streit and Bolam, (2000) Neuroscience 101:1037-1051.

Afferents of dopamine neurons

Glutamatergic afferents

STN

PPN

Cortex

Amygdala

Bolam, Francis and Henderson (1991) Neuroscience 41:483-494

Ross, Mena-Segovia, Magill and Bolam unpublished
Afferents of dopamine neurons the
substantia nigra pars compacta

Superior colliculus

5-Hydroxytryptamine: Raphe

Noradrenaline: LC

Peptides

Dopamine
Afferents of dopamine neurons the
substantia nigra pars compacta
SUMMARY
Dopamine neurons receive a complex
afferent innervation mainly from
GABAergic and glutamatergic terminals
Ionotropic GABAA and glutamate
receptors are mainly located at synapses
Metabotropic receptors are located at
synapses and extrasynaptic sites
GABAergic innervation mainly from
St, GP and SNR
Glutamatergic innervation mainly from
STN and PPN
Other functionally important afferents
The complexity and simplicity of dopamine neurons

Organisation of dopamine neurons at level of SNC

(normality)
Projections of dopamine neurons (complexity)
Responses of dopamine neurons (simplicity)
The effects of dopamine neurons (subtlety)
Projections of dopamine neurons

Striatum

GP
SNC

SNR

DAT immunolabelling
Projections of dopamine neurons DAT immunolabelling
Striatum

GP
Striatum

Globus pallidus
 The projections of
dopamine neurons

Topography of the projection

medial to lateral/limbic to motor

Dorsal and ventral tier

Striatonigral projection
patch and matrix

Based on retrograde and

anterograde labelling

Bentivoglio and Morelli ( 2005) Handbook Chem. Neuroanat 21:1-107

 The projections of
dopamine neurons

Information flow from

frontal cortex through
the striatonigral circuit

Haber, et al., 2000 J. Neurosci.

The projections of dopamine neurons

Data from a variety of sources

including immunocytochemistry and
single cell filling indicate that
dopaminergic neurons innervate
other regions of basal ganglia

GPe Rodrigo et al 1998

Smith & Kieval 2000
Gauthier et al 1999
Prensa & Parent 2001
The projections of dopamine neurons

Data from a variety of sources

including immunocytochemistry and
single cell filling indicate that
dopaminergic neurons innervate
other regions of basal ganglia

GPe Rodrigo et al 1998

Smith & Kieval 2000
Gauthier et al 1999
Prensa & Parent 2001
 The projections of dopamine neurons
GPe contains DAT-positive axons and boutons

Striatum

GP

Prime, Boyes, Magill and Bolam unpublished

 The projections of dopamine neurons

TH immunolabelling Rodrigo et al 1998; Prime, Boyes, Magill and Bolam unpublished

The projections of dopamine neurons
STN contains TH-positive and DA-positive axons and boutons

Cragg, Baufreton, Xue, Bolam & Bevan 2004 Eur. J. Neurosci. 20:1788-802
The projections of dopamine neurons
DA is released from STN upon electrical stimulation

Cragg, Baufreton, Xue, Bolam & Bevan 2004 Eur. J. Neurosci. 20:1788-802
The projections of dopamine neurons
DA affects the firing of STN neurons

Cragg, Baufreton, Xue, Bolam & Bevan 2004 Eur. J. Neurosci. 20:1788-802
 The projections of dopamine neurons

In addition to innervation of the striatum, dopamine

neurons innervate other regions of the basal ganglia

Loss of dopamine at the level of the GP and/or the STN

may account for some of the changes in Parkinsons
disease

Degree of collateralization of individual neurons?

Single cell filling studies

 The projections of dopamine neurons

Small deposits of
anterograde tracer
in SNC

Single axons traced to

forebrain
Characterize the axonal
arborization of single
neurons
Respect patch matrix

Prensa and Parent (2001) J Neurosci 21:7247-7260

 The projections of dopamine neurons

Several classes of neurons

based on location and
projection

Poor respect for the patch/

matrix organisation

High degree of
collateralization

Problems with the technique

DA or not
size of arborization

Based on quantitative
anatomical data the
arborization of a single
dopamine neuron is massive
Prensa and Parent (2001) J Neurosci 21:7247-7260
 The projections of dopamine neurons

Number of synapses formed by dopamine neurons

15000 spines per spiny neuron (Wilson)
4% of spines receive symmetrical synapses (Wilson)
Assume half are dopamine terminals and half of dopamine terminals are on
spines
7.2 x 103 dopamine neurons in SNC (Oorschott)
2.78 x 106 spiny neurons in striatum(Oorschott)
THEREFORE
15000 x 0.02 x 2 x 2.78 x 106 / 7.2 x 103= 231 x 103 synapses per neuron
Freund et al 40% spines receive dopamine input which accounts for
60% of terminals
THEREFORE
15000 x 0.4 x 2.78 x 106 / 7.2 x 103 +40% = 3.24 x 106 synapses per neuron
 369,881

Wickens and Arbuthnott (2005) Handbook Chem. Neuroanat 21:199-236

 Neurons of the globus pallidus

Kita and Kitai (1994) Brain Res, 636, 308-309

Bevan, Booth, Eaton, and Bolam. (1998) J. Neurosci. 18, 9438-9452
The complexity and simplicity of dopamine neurons

Number of synapses formed by globus pallidus neurons

Striatum 790
GP 264 or 581
SNR 36-209
SNC 11-28
EP approx 100
STN 41-274

Total Max around 1982 synapses per neuron

Kita and Kitai (1994) Brain Res, 636:308-309

Bevan, Booth, Eaton, and Bolam. (1998) J. Neurosci. 18: 9438-9452
Sadek, Magill and Bolam unpublished
The complexity and simplicity of dopamine neurons
Number of synapses formed by striatal neurons

around 300 synapses per neuron

Data from: Kawaguchi et al 1990 J. Neurosci. 10:3421-3438

Wu et al 2000 Neurosci Res 38:49-62
 PV-positive GABAergic interneuron
Approx. 5000 boutons

Tepper and Bolam (2004 ) Current Opinion Neurobiol.

The complexity and simplicity of dopamine neurons

Individual dopamine neurons give rise to a massive

number of synapses in the striatum

Non-synaptic release sites, collaterals to other regions

Difficulties of control of the whole arborization

(branch point failures, local control of release at level of terminals,
metabolic support for such a large arborization)

Different to other neurons in the basal ganglia

The complexity and simplicity of dopamine neurons

Juxtacellular labelling of individual

dopamine neurons in vivo, chemical
characterisation and re-construction

Quantitative data on degree of

collateralization, numbers of
synapses formed and the nature and
pattern of innervation of their
postsynaptic targets

Boyes, Magill and Bolam unpublished

Ungless, Magill and Bolam 2004 Science 30:2040-2042
The complexity and simplicity of dopamine neurons

Organisation of dopamine neurons at level of SNC

(normality)
Projections of dopamine neurons (complexity)
Responses of dopamine neurons (simplicity)
The effects of dopamine neurons (subtlety)
Activity of dopamine neurons
The complexity and simplicity of dopamine neurons

Organisation of dopamine neurons at level of SNC

(normality)
Projections of dopamine neurons (complexity)
Responses of dopamine neurons (simplicity)
The effects of dopamine neurons (subtlety)
Spiny projection neurons
Major target of the nigrostriatal projection

Freund, Powell & Smith (1984)

The complexity and simplicity of dopamine neurons

Major target of dopamine

terminals is the spines of MSNs

Modulate the flow of cortical

information

Also innervate striatal

interneurons

Actions mediated through at

least two classes of receptors

Smith, Bennett, Bolam, Parent and Sadikot (1994) J. Comp. Neurol. 344:1-19
The complexity and simplicity of dopamine neurons

Dopamine receptors located at synaptic and

extrasynaptic sites
Technical issues relating to receptor
immunolabelling
Yung, Bolam, Smith, Hersch, Ciliax & Levey (1995). Neuroscience. 65 :709-730
The complexity and simplicity of dopamine neurons

Dopamine system is so remarkable

because of the plasticity of the
corticostriatal synapse

Dopamine modulates the flow of

cortical information

Plasticity of the corticostriatal

synapse is often dependent on
dopamine

Brought about by a variety of

mechanisms including LTP, LTD,
changes in excitability, receptor
interactions, signalling pathways
and gene regulation
1 Figure 4. Effect of dopamine on corticostriatal synapses. Effect of dopamine on activity-
2 dependent synaptic plasticity in the corticostriatal pathway. A. A conjunction of cortical
3 presynaptic activity and striatal postsynaptic activity, leads to long-term depression (LTD) in the
4 absence of a dopamine pulse. B. A conjunction of cortical presynaptic activity and striatal
5 postsynaptic activity, leads to long-term potentiation (LTP) if preceded by a dopamine pulse. C.
6 The same conjunction of cortical presynaptic activity and striatal postsynaptic activity, leads to
7 LTD if the dopamine pulse is delayed.
Bolam, Bergman, Graybiel, Kimura, Plenz, Seung, Surmeier, and Wickens (2005) In press.
The complexity and simplicity of dopamine neurons

Dopamine neurons have a complex organisation and afferent input

at level of SNC (normality)
The projections of dopamine neurons are extremely complex and
massive (complexity)
Responses of dopamine neurons are relatively simple and uniform
(simplicity)
The effects of released dopamine at the level of the striatum (other
regions of BG/ctx) are subtle and complex and enable dopamine to
have profound effects on behaviour (subtlety)
The challenge we have set ourselves in DOPANET is to understand
one of the most complex neurons in the brain and one of the most
complex neurotransmitters
The complexity and simplicity of dopamine neurons

Thanks to:
Peter Magill
Yoland Smith
Justin Boyes
Mark Ungless
Fumino Fujiyama
Tracey Chatha
Hana Ross
Juan Mena-Segovia
Mark Bevan
Stephanie Cragg
Bolam, Bergman, Graybiel, Kimura, Plenz, Seung, Surmeier, and Wickens (2005) In press.