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Dendritic spines, Rac 1

and Neuropathic Pain after Spinal Cord Injury
Dendritic spines are small protrusions from
neuronal dendrites that typically receives input from the
presynaptic component of the excitatory synapses in the NMDAR
central nervous system. They are located on the den- NM
DA DA
drites. The most common morphological type is com- NM NM
posed of a bulbous head and a thin neck. The head is con-
DAR DA
nected with the dendrite through the neck. There is a po- N M GluR R
tential relation between spine shape and synaptic func- hilin
spinop
u


CaMKII
tion, since morphological rearrangements of spines have P
+
Ca2+
p i n
been found in vitro and in vivo [1,2]. In samples of spines
Dendritic Spine
PIIa LCF PSD-95
& L. S
from layer 2/3 pyramidal neurons from mouse primary ROCK CaMKII + ars i l a
visual cortex, using first Golgi impregnations and then A  C. B

RhoA
y


+ b

A
A

A
gold-toning and serial thin section electron microscopy, 5 GTP
pt
D-9 Tiam 1 integrin
ce
no detectable correlations between spine head volume PS
n
Par3
and spine neck length were found [3]. The area of the
SAPAP/GKAP P
P
&co
PSD
A
RhoA
s i gn
de

A
A

A
is correlated to the spine head volume and neck GDP ProSAP/Shank
hilin FAK4
diameter and it is uncorrelated with the spine neck
length. In rat, in CA1 pyramidal cells, the volume of the PAK
spinop
PAK PIX + + RAC1
spine head was reported to be proportional to the post-
+
GDP

membran
+
R
synaptic density (PSD) area and to the number of presyn-
+
POR 1

Gro
aptic vesicles [4]. The spine neck length is correlated to DA Ca2+/Calmodulin
PIIa
RAC1

wt
NM

e ruffling

hF
-
the time constant of calcium compartmentalization [3] LIMK
p140SRA-1

act
+
GTP

-
and also proportional to the filtering of electrical poten-

or
ROCK

R
tials and it may be involved in calcium dependent learn-

ece
AP
ing rules. There is no correlation between head volume GK myosin

pto
/ p70S6K
PAP k binding activation translation
and neck length, although there is a weak correlation A an

r
S cofilin
/Sh PIX cofilin P
between head volume and neck diameter [3]. In neocorti- AP
oS K
cal pyramidal neurons spines that were further away from
the soma were longer and had larger heads. In CA1 pyra- Src
Pr PA
myosin
P
+ IQGAP
RAS
GTP
P32/CDK5R1 PIP5K
midal neurons spines located in the distal portions of the PAK actin polymerisation
apical dendrite had larger heads [6]. Similar effects were + in u
p
-
CN/PP2B
found in Golgi-impregnated CA1 pyramidal neurons,
debrin binding activation PLD 1
L. S
cortactin + MLCK l a &
Dendritic spines

P67 (Phox)
albeit not in neocortical pyramidal cells from layers 2/3, 4,
gelsoline actin polimerisation
a r s i
5, and 6 [7]. In spines from layer 2/3 pyramidal neurons, no
Arp2/3 PAR-6
cell-cell adhesion
ARHGEF6
C .B
significant relation was found between distance from the
by
Ca2+
soma and spine head volume, total spine volume and PSD
TrkB + N-WASP
e pt
area. There is correlation between spine head volume and
debrin phosphatidic acid level
o n c
+ P NADPH oxidase
& c
the area of the PSD. It may be a correlation between the myosin PARD3 PI3K
s i gn
volume of the spine head and the synaptic strength or it
de

design & concept by C. Barsila & L. Spinu

MAPK RAC1 aPKCs RAC1
may be linked to the release probability [5]. There is no ROS
Raf 1
clear correlation between the spine head volume and GDP
spine neck length. There is a lack of correlation between GTP
RAC1
+
Par3

head volume and neck length (since the PSD area is corre-
+ kalirin kalirin
NF-KB
+

MTOC orientation
lated with the head volume) [5]. The PSD area is itself pro-
1

GTP
Tiam

Tiam 1
P

portional to the number of postsynaptic receptors [8]. The

+
volume of the spine head is likely to be directly propor- GDP
KII

http://www.thalamus.ro
CaM

tional to the average reliability and strength of its syn- cell proliferation
-95

Posh
apse. CA1 pyramidal neurons show a larger spine size
RAC1
PSD

with increasing distance from the soma [7, 6], as if synap- actin regulatory pathways mediated
tic weight is compensating for the dendritic electrotonic MEK1/2
IRSp53 by non-glutamate receptors
filtering [5]. CYFIP2
Arp2/3 has actin-polymerizing activity.
1. Dunaevsky A, Tashiro A, Majewska A, Mason C, and Yuste R. Develop- Hspc300
mental regulation of spine motility in the mammalian central nervous Abl2 WAVE N-Wasp has actin-polymerizing activity.
WAVE genes MLK3
system. Proc Natl Acad Sci USA 96: 13438–13443, 1999
genes MEKK1 Both of them (Arp2/3 and N-Wasp) depend on
2. Fischer M, Kaech S, Knutti D, and Matus A. Rapid actin-based plasticity NcKAP1
in dendritic spines. Neuron 20: 847–854, 1998. cortactin phosphorylation levels. This levels
3. Barbara Calabrese, Margaret S. Wilson and Shelley Halpain, Develop- WAVE are controlled in a TrkB and Src dependent
ment and Regulation of Dendritic Spine Synapses, Physiology 21:38-47, MKK4/7 active
2006 manner. Kalirin is recruited and activated to
4. Harris KM and Stevens JK. Dendritic spines of CA 1 pyramidal cells in the spines by EphB. Kalirin through Rac and PAK
rat hippocampus: serial electron microscopy with reference to their
biophysical characteristics. J Neurosci 9: 2982–2997, 1989. JNK1 leads to activation of myosin.
5. Jon I. Arellano, Ruth Benavides-Piccione, Javier DeFelipe,and Rafael
Arp2/3 actin regulatory proteins in spines. The
Yuste Ultrastructure of dendritic spines: correlation between synaptic and
spine morphologies, Frontiers in Neuroscience. (2007) vol. 1,
actin severing activity of cofilin is dependent on
iss. 1,131-143
genes
the balance existing between kinases and phos-
6. Megias, M., Emri, Z., Freund, T. F., Gulyas, A. I. (2001). Total number and phatases. - LIMK and CN/PP2B. Cofilin binds to
distribution of inhibitory and excitatory synapses on hippocampal CA1
pyramidal cells. Neuroscience 102, 527–540. actin and affects the filament structure. At this
7. Konur, S., Rabinowitz, D., Fenstermaker, V., Yuste, R. (2003). Systematic moment the debrin afinty for actin is lowered.
regulation of spine head diameters and densities in pyramidal neurons
from juvenile mice. J. Neurobiol. 56, 95–112. Actin regulationg pathways under the control of ionotropic glutamate receptor. Debrin has a stabilizing effect on actin. Debrin
8. Nusser, Z., Lujan, R., Laube, G., Roberts, J., Molnar, E., Somogyi, P. (1998). prevents actin reorganization. The reorganization
Cell type and pathway dependence of synaptic AMPA receptor number RhoA (activated) interacts with NMDARs. The effect is the activation of the ROCK/PII complex. The of actin is due to myosin binding to acting
and variability in the hippocampus. Neuron 21, 545–559.
result is a stable actin after ROCK/PII activation. High levels of CA2+ induce CaMKII-dependent phos- filaments and by interacting with gelsolin.
phorylation of spinophilin. This detaches the spinophilin from the actin and sends it to the mem- Myosin stabilizes actin and contracts F-actine.
brane. Here spinophilin interacts with Lcf. After this interaction Lcf activates RhoA. The actin-severing PAK triggers myosin motor activity. Gelsolin caps
activity of cofilin is controlled by different kinases and phosphatases. LIMK has a negative regulator the barbed ends of actin. In this way the actin
effect on cofilin activity. Activation of LIMK depends on Rac-1, through its effector - PAK. NMDA polimerization is possible. This role of gelsolin is
stimuli increases Rac-1 activity (local) through Rac1 - GEFs PIX and Tiam 1 which increases the activity Ca2+ dependent.
of cofilin, wich can lead to higher actin turnover rates.
Vanessa Schubert and Carlos G. Dotti, Vanessa Schubert and Carlos G. Dotti,
Transmitting on actin: synaptic control of Transmitting on actin: synaptic control of
dendritic architecture, Journal of Cell dendritic architecture, Journal of Cell
Science 120, 205-212 Science 120, 205-212
Dendritic Spines Dendritic Spines

Dendritic spines Dendritic spines Dendritic spines

Localized increases in synaptic strength constitute a synaptic basis for learning and
memory in CNS.
5 350 One of the key regulators of the actin cytoskeleton is
Spinal cord injury (SCI) could determine dendritic spines remodeling and can con- the Rho family of GTPases . The
spines/10microm. dendrite

tribute to neuronal hyperexcitability and neuropathic pain through synaptic changes. Rho GTPases function as molecular switches to turn on
Synaptic plasticity induced by SCI may appear in the spinal cord dorsal horn and may or off downstream biochemical pathways depending
2,5 on the stimuli [1]. This GTP-ases are under control of
0 contribute to pain maintenance [1,2]. SCI increases Rac1 mRNA expression, which
proteins such as the guanine nucleotide exchange
remains elevated for up to 3 months [3[. A role of Rac1, in neuropathic pain after SCI it factors (ex:Kalirin-7) and the GTPase-activating pro-

u
is not studied enough. Rac1 can modulate dendritic spine morphology and function

n
teins. Rac and Cdc42 promote neurite outgrowth, RhoA
[4, 5]. Andrew M. Tan et al (2008) applied the Rac1-specific inhibitor NSC23766 in order stimulates retraction. The balance of these opposing
activities of the different Rho GTPases regulates some

. S p i
L
to study the effect of synaptic remodeling in neuropathic pain after SCI. Rac1-specific

a&
0 functions and the morphology of neurons.
0 350 inhibitor NSC23766 blocks guanine exchange factors (GEFs), Trio and Tiam1. Inhibi-

l
GTPases functions: cell movement and motility, tran-
distance from the soma tion of the Rac1 signaling cascade ameliorated the development of abnormal spine scription, cell growth and proliferation, as well as cell

a r s i
B
morphologies, reduced neuronal excitability, and normalized nociceptive thresholds. cycle progression. Members of the Rho Molecules in
GTPase family have two states: GTP- and GDP-bound

C .
by
[6] PSD-95 expression is a marker of sites of synapse plasticity. Expression of PSD-95 is states. The GTPases are inactivated when the bound

t
increased significantly in injured spinal cord tissue compared with uninjured controls GTP is hydrolyzed to GDP. Rho GTPases have intrinsic
[6]. NSC23766 treatment reduces PSD-95 levels below that of uninjured levels . Cortac- GTPase activity but the hydrolysis is slow. The Rho
GTPases are normally present in cytoplasm, kept here

n ce p
o
tin levels did not significantly change after NSC23766 treatment compared with intact

c
by RhoGDI until a stimuli is applied to the cell. Well

&
animals. known members of Rho GTPases are RhoA, Cdc42 and
Dendritic spine density increases after SCI. In SCI plus veh animals (0.9% saline), the
Rac1 [1]. The effector proteins downstream of Rac1
s i gn
e
density of spines significantly shift toward the cell body compared with the spine den-

These graphics are orientative and represent
the pattern found by Andrew M. et al 2008. For the
original data see Andrew M. Tan, Severine Stamboulian,
Yu-Wen Chang, Peng Zhao, Avis B. Hains,2 Stephen G. Waxman,
and Bryan C. Hains, Neuropathic Pain Memory Is Maintained by
Rac1-Regulated Dendritic Spine Remodeling after Spinal Cord
Injury, The Journal of Neuroscience, 28(49):13173–13183 • 13173
d
in lamellipodia formation are mainly the WAVE subfam-
sity distribution in intact animals. An increase in spine density and redistribution of ily of the WASP proteins. POR1 may also be involved in
spine location relative to the cell body, and increases in spine length and head diameter this process [2,3,4]. N-WASP mediates the link between
Cdc42 and the Arp2/3 proteins in actin polymerization,
after SCI) occurs after SCI in dorsal horn neurons. and participates in the formation of filopodia [2, 5] .
WAVE1 is a downstream effector of Rac1. It is respon-
Rac1 could be interpreted as critical in maintaining neuropathic pain through its regu- sible for the number of dendritic spines in the neurons.
lation of dendritic spines. WDR (Wide-Dynamic Range ) neurons exhibit hyperexcitabil- Phosphorylation of WAVE1 by the cyclin-dependent
kinase 5 (Cdk5) inhibits WAVE1’s activity and thus limits
ity in response to evoked-innocuous and noxious stimulation after SCI. Also increased its capacity to regulate Arp2/3-dependent actin polym-
sensitivity to mechanical stimuli and reduced thermal nociceptive thresholds are found erization [1,6] . Cdk5 and its regulator p35 have also
after SCI. been shown to interact with both Rac1 and PAK leading
NSC23766 is specific for Rac1 and does not interfere with cdc42 or RhoA-GEF or Rac1 to downregulation of PAK activity [7]. It has long been
established that the downstream effects of RhoA and
Neuropathic Pain after Spinal Cord Injury
binding to its effector PAK1. Andrew M. Tan et al (2008) conclude that a post-SCI shift in Cdc42/Rac can be antagonistic to one another in cells
the number of mature spines may contribute to the strengthening of synaptic inputs, [8] . Cdc42 and Rac are required for neurite formation
while dominant negative Cdc42 and Rac1 have been
enhanced transmission fidelity, and potentiation of electrical transduction, which to-
found to inhibit neurite outgrowth in N1E115 cells [9].
gether may lead to a pathological amplification of sensory information after SCI [6]. Strong Rac1 and Cdc42 activities have also been local-
ized to the tips of the growin neurites in PC12 cells
1 .Woolf CJ, Shortland P, Coggeshall RE (1992) Peripheral nerve injury triggers central stimulated with nerve growth factor (NGF) [1,10] . The
RhoA-induced neurite retraction was found to be medi-
sprouting of myelinated afferents. Nature 355:75–78. ated by the actions of ROK. Studies on primary neurons
2. Kim BG, Dai HN, McAtee M, Vicini S, Bregman BS (2006) Remodeling of synaptic struc- have also confirmed the findings that Cdc42 and Rac1
tures in the motor cortex following spinal cord injury. Exp Neurol 198:401– 415. generally enhance neurite formation and outgrowth
3. Erschbamer MK, Hofstetter CP, Olson L (2005) RhoA, RhoB, RhoC, Rac1, Cdc42, and whereas RhoA activity inhibits these activities [1]. How-
ever, recent data have indicated that it is the balance of
Tc10 mRNA levels in spinal cord, sensory ganglia, and corticospinal tract neurons and Rho GTPase activities that is important in the regulation
long-lasting specific changes following spinal cord injury. J Comp Neurol 484:224 –233. of neurite outgrowth. Too much or too little Rac1 activ-
4. Nakayama AY, Harms MB, Luo L (2000) Small GTPases Rac and Rho in the maintenance ity reduces neurite outgrowth.
of dendritic spines and branches in hippocampal pyramidal neurons. J Neurosci

u
1 Cheng-Gee Koh, Rho GTPases and Their Regulators in Neuronal Func-

i n
20:5329 –5338. tions and Development, Neurosignals 2006–07;15:228–237

p
2 Miki H, Suetsugu S, Takenawa T: WAVE, a novel WASP-family protein

S
5. Wiens KM, Lin H, Liao D (2005) Rac1 induces the clustering of AMPA receptors during
.
involved in actin reorganization induced by Rac. EMBO J 1998; 17:

L
6932–6941.
spinogenesis. J Neurosci 25:10627–10636.

a&
3 Miki H, Yamaguchi H, Suetsugu S, Takenawa T: IRSp53 is an essential

l
6. Andrew M. Tan, Severine Stamboulian, Yu-Wen Chang, Peng Zhao, Avis B. Hains,2 Ste-
i
intermediate between Rac and WAVE in the regulation of membrane

r s
ruffling. Nature 2000; 408: 732–735.

a
phen G. Waxman, and Bryan C. Hains, Neuropathic Pain Memory Is Maintained by Rac1- 4 Van Aelst L, Joneson T, Bar-Sagi D: Identification of a novel Rac1-

. B
interacting protein involved in membrane ruffling. EMBO J 1996; 15:
Regulated Dendritic Spine Remodeling after Spinal Cord Injury, The Journal of Neuro-

C
3778–3786.

by
science, 28(49):13173–13183 • 13173 5 Rohatgi R, Ma L, Miki H, Lopez M, Kirchhausen T, Takenawa T, Kirsch-

t
ner MW: The interaction between N-WASP and the Arp2/3

e p
complex links Cdc42-dependent signals to actin assembly. Cell 1999;

c
Rac1 (Ras-related C3 botulinum toxin sub- 97: 221–231.

o n
6 Kim Y, Sung JY, Ceglia I, Lee K-W, Ahn J-H, Halford JM, Kim AM, Kwak
strate 1) is a small (~21 kDa) GTPase, and is a

c
SP, Park JB, Ho Ryu S, Schenck A, Bardoni B, Scott JD, Nairn

&
member of the Rac subfamily of the family AC, Greengard P: Phosphorylation of WAVE1 regulates actin polymer-

n
ization and dendritic spine morphology. Nature 2006;
Rho - family of GTPases. It is encoded by the

s i g
442: 814–817
gene RAC1.

e
7 Nikolic M, Chou MM, Lu W, Mayer BJ, Tsai LH: The p35/Cdk5 kinase is

d
www.thalamus.ro a neuron-specific Rac effector that inhibits Pak1 activity. Nature 1998;
Dendritic spines contain a cytoskeleton com-
The Thalamus
395: 194–198.
posed mostly of filamentous actin (F-actin) which 8 Kozma R, Ahmed S, Best A, Lim L: The Rasrelated protein Cdc42Hs
and bradykinin promote formation of peripheral actin microspikes and
determines the shape and stability/motility of filopodia in Swiss 3T3 fibroblasts. Mol Cell Biol 1995; 15: 1942–1952.
neuroscience & medicine spines. Spines have a small amount of intermedi- 9 Sarner S, Kozma R, Ahmed S, Lim L: Phosphatidylinositol 3-kinase,
Cdc42, and Rac1 act downstream of Ras in integrin-dependent neurite
ate filaments and microtubules, ellements that outgrowth in N1E-115 neuroblastoma cells. Mol Cell Biol 2000; 20:
are present in large mumber in the dendritic shaft 158– 172.
. 10 Aoki K, Nakamura T, Matsuda M: Spatiotemporal regulation of Rac1
and Cdc42 activity during nerve growth factor-induced neurite
Barbara Calabrese, Margaret S. Wilson and Shelley outgrowth in PC12 cells. J Biol Chem
Halpain Development and Regulation of Den- 2004; 279: 713–719.

dritic Spine Synapses, Physiology 21:38-47, 2006

This poster does not intend to be an “authoritative” article. It represents our interest in research http://neuroscience-bucharest.blogspot.com
and also it represents our interest in medical graphic design. You are not allowed to sell or use this
poster or parts of it in any circumstances. You can use this poster for personal purposes or for http://www.thalamus.ro
educational purposes. If you use this poster please link back or leave a comment on
C design & concept by C. Barsila & L. Spinu
http://neuroscience-bucharest.blogspot.com or http://www.thalamus.ro 2009
medical students