spinal cord injured monkeys discussion - unifr.ch...f3 –layerv spinal cord injured macaques f3...
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Materials and methods2• Cell density = Hemisphere cell number/ Delimited area (µm ) x Sectrion›s
thickness (µm)
Interhemispheric difference of cell density (IDCD) was calculated by
subtracting the cell density of the directly affected hemisphere by the cell
density of the intact hemisphere.
• Handedness index (HI) post lesion = H-I/H+I from H=score of the healthy
hand on the Brinkman board task and I=score of the «affected» hand on
the Brinkman board task.
• Positive IDCD means more SMI-32 neurones visible in the lesioned
hemisphere. Negative is the reverse.
A unilateral lesion of the hand representation in the primary motor cortex or a partial hemisection of the cervical cord
in macaques affects the interhemispheric ratio of SMI-32 stained neurons in premotor cortical areas
Schmidlin E., Colangiulo R., Contestabile A., Kaeser M., Savidan J., Wyss A.-F., Hamadjida A., Rouiller E. M. Department of Medicine, Neurosciences, University of Fribourg, Swi�erland
III IV V
-0.0008
-0.0006
-0.0004
-0.0002
0
0.0002
0.0004
0.0006
0.0008
0.001
Pre-SMA(F6) SMA-proper (F3) PMv-r (F5)
Intact M1 injured monkeys Spinal cord injured monkeys
Introduction
The primary motor cortex (M1) of primates is interconnected with premotor areas, in both hemispheres, such as the rostral part of the ventral premotor cortex (PMv-r or F5) and the supplementary motor area (SMA: pre-SMA = F6 and SMA-proper = F3)). The working hypothesis was that after a unilateral lesion of M1,
there is a change of the SMI-32 positive cells ipsilesionally in PMv-r and SMA. To assess possible effects of a permanent unilateral lesion of M1 (hand region) or of an hemisection of the spinal cord (C7/C8 level) on PMv-r and SMA, a histological analysis of several macaques’ brain was performed by comparing between
the 2 hemispheres the neuron density of layers III and V in these premotor areas.
In this study, three subpopulations of adult animals were analyzed: (i) ; (ii) (two of them were treated with anti-Nogo A antibody); (iii) (three of them were treated with anti-Nogo A antibody).five intact control animals seven animals with a unilateral lesion of M1 nine macaques with a unilateral lesion of the spinal cord
Figure 2. Two photomicrographs of SMI-32 stained cells in SMA at 40x
magnification (image above, scale bar 200 µm) and 200x (image below,
scale bar 100 µm). The SMA delimitation and the layer’s parcellation
are shown.
SMA
PMv-r Mk-CE
Mk-CG
Figure 1. Schematic view of a cortical lesion (red area, left panel)
and a spinal lesion (blue area, right panel).
Results
• Intact monkeys have a li�le interhemispheric difference of cellular distribution in layer III and V of SMA and
PMv-r.
• In several injured monkeys, an interhemispheric significant cell density difference was observed (Figures 3 and
5).
• The SMI-32 cell density is more affected in F3 than in F6 (Figure 5), as expected.
• The treatment with anti-Nogo-A antibody does not seem to affect the cell density of the premotor areas.
• Five out of seven M1 injured monkeys and four out of nine spinal cord injured monkeys have a greater layer V
SMI-32 cell density in SMA in «intact» hemisphere.
• Age and functional recovery show some correlation with SMI-32 cell›s interhemispheric distribution.
Discussion
After cervical cord lesion (SCI) or M1 lesion, there was a
change of SMI-32 expression in layer V of SMA-proper, as
expressed by significant positive or negative IDCD in 5 M1
lesion monkeys and 4 SCI monkeys. In contrast, 2 out of
seven M1 lesion monkey and 5 out of 9 SCI monkeys did not
show a significant IDCD, like intact animals. Largely
comparable changes in PMv-r were observed.
Figure 8. IDCD in layer V of SMA plo�ed as a function HI post lesion. M1 injured
monkeys in red and spinal cord injured macaques in orange. The diamonds indicate
the anti-Nogo-A antibody treated monkeys.
Figure 6. IDCD in layer V of SMA was plo�ed in function of the age at sacrifice (upper panel), the extent of the lesion (middle panel) and the duration of the
recovery phase (lower panel). Intact monkeys in green, M1 injured monkeys in red and spinal cord injured macaques in orange. The diamonds indicate the anti-
Nogo-A antibody treated monkeys.
Figure 7. IDCD in layer V of SMA plo�ed as a function of to the functional recovery.
M1 injured monkeys in red and spinal cord injured macaques in orange. The
diamonds indicate the anti- Nogo-A antibody treated monkeys.
Figure 5. IDCD in layer V of F6 (dark blue), F3 (light blue) and PMvr (sky-blue) in all animals involved in the study.
Figure 3. IDCD in layer III of F6 (dark blue) and F3 (light
blue), showing a difference in M1 injured monkeys but not in
spinal cord injured monkeys. As compared, SCI has no
effect on layer III , in contrast to M1 lesion.
Intact M1 injured monkeys Spinal cord injured monkeys
-0.001
-0.0005
0
0.0005
0.001
0.0015
Mk-IC
Mk-RO
Mk-BI
Mk-SL
Mk-CG Mk-CGa Mk-AG
Pre-SMA(F6) SMA-proper (F3)
3ID
CD
(ce
ll n
umbe
r/µ
m)
CG
CP
AP
CS
CGa
CH
AM
AG
CC
R² = 0.1703
R² = 0.8734
0 5 10 15 20 25 30 35 40 45 50
Duration recovery phase (day)
GEBI
JU
VA
ROCE
SL
R² = 0.5634
R² = 0.2989
-0.0008
-0.0006
-0.0004
-0.0002
0
0.0002
0.0004
0.0006
0.0008
0.001
10 30 50 70 90 110
Duration recovery phase (day)
RO
GEBI
JU
CE
SL
VA
R² = 0.2727
-0.0008
-0.0006
-0.0004
-0.0002
0
0.0002
0.0004
0.0006
0.0008
0 20 40 60 80 100 120
Volume of the lesion (mm3)
M1 injured macaquesF3 – Layer V
Spinal cord injured macaquesF3 – Layer V
IZIRIE
RO
GEBI
JU
CE
R² = 0.5716
-0.001
-0.0008
-0.0006
-0.0004
-0.0002
0
0.0002
0.0004
0.0006
0.0008
1800 2000 2200 2400 2600 2800
Age at sacrifice (day)
IZIR
IEIC
CG
CH
CGa
AM
CP
AP
AG
CS
CC
R² = 0.4117
1200 1700 2200 2700 3200 3700 4200
Age at sacrifice (day)
CG
CH
CGa
AM
CP
AP
AG
CS
CC
R² = 0.255
0 10 20 30 40 50 60 70 80 90 100
Proportion of hemisection (%)
3ID
CD
(ce
ll n
umbe
r/µ
m)
3ID
CD
(ce
ll n
umbe
r/µ
m)
3ID
CD
(ce
ll n
umbe
r/µ
m)
GEBI
JU
VA
CG
CP
AP
CS
CGa
ROCE
SLCH
AM
AG
CC
R² = 0.3917
R² = 0.088
-0.0008
-0.0006
-0.0004
-0.0002
0
0.0002
0.0004
0.0006
20 30 40 50 60 70 80 90 100 110 120
Functional recovery (%)
3ID
CD
(ce
ll n
um
ber
/µm
)
GEBI
JU
VA
CG
CP
AP
CS
CGa
RO
CE
SLCH
AM
AG
CC
R² = 0.4569
R² = 0.0262
-0.0008
-0.0006
-0.0004
-0.0002
0
0.0002
0.0004
0.0006
0.0008
0.001
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7
HI post lesion
3ID
CD
(ce
ll n
umbe
r/µ
m)
3ID
CD
(ce
ll n
umbe
r/µ
m)
Figure 4. Graphics that show the rostro-caudal gradient of cell density per histological section in layer V. An example for SMA and PMv-r
is presented for an intact (above) and an M1 injured (below) animals.
0.00000
0.00020
0.00040
0.00060
0.00080
0.00100
0.00120
Rostral Caudal
PMv-r Layer V, Mk-IU
Left hemisphere Right hemisphere
0
0.0001
0.0002
0.0003
0.0004
0.0005
0.0006
0.0007
0.0008
Rostral Caudal
SMA - Layer V, Mk-IE
Left hemisphere Right hemisphere
0.00000
0.00020
0.00040
0.00060
0.00080
0.00100
0.00120
Rostral Caudal
PMv-r Layer V, Mk-RO
Left hemisphere Right hemisphere
0
0.0002
0.0004
0.0006
0.0008
0.001
0.0012
0.0014
0.0016
0.0018
Rostral Caudal
Left hemisphere Right hemisphere
M1 in
jure
d M
aca
que
Inta
ct M
aca
que
3ID
CD
(ce
ll n
umbe
r/µ
m)
3ID
CD
(ce
ll n
umbe
r/µ
m)
3ID
CD
(ce
ll n
umbe
r/µ
m)
3ID
CD
(ce
ll n
umbe
r/µ
m)
SMA - Layer V, Mk-RO
Grant sponsors: Swiss National Science Foundation, Grants No. 31-61857.00, 310000-110005, 31003A- 132465 (EMR), PZ00P3_142258 (ES).The Swiss Primate Competence Centre for Research (SPCCR).
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