Differential metal repartition in lesion and control ventral horns of

young adult rat spinal cord

Tina Perkins1, Seritta Hill1, Stefan Vogt2, Christine Gerin1,3
(1) Chicago State University, (2) APS-Argonne Natl Laboratory, (3) University of Illinois at Chicago

Abstract
Literature shows that neurodegenerative diseases such as
Amyotrophic Lateral sclerosis (ALS) are correlated to
metal (Fe, Cu, Zn) accumulation. We hypothesize that
metal accumulation might be triggered by injury to the
central nervous system. After injury, gray matter shows
less protection than white matter. We aim to quantify
metal repartition within the T9 segment of the gray
matter of the spinal cord and to compare ipsi- and
contralateral side to the hemisectioned spinal cord.
Differential mineral accumulation was quantified and
analyzed. Fresh rat spinal cords were excised, fast
frozen, cryostat sectioned and freeze-dried. At the
BioCat beam line, sample sections were scanned onto
the X-ray fluorescence microprobe using continuous
scanning (20m scans). Our results show that Fe is more
accumulated in the dorsal part of the ventral horn. Cu
repartition occurs all over the ventral horn, more
ventrally than Fe. Zn accumulation occurs more
ventrally than Fe and Cu. Average amounts, in the
control side, for Fe, Cu, Zn are respectively (g/cm2)
1.50.05; 0.170.01; 0.460.01. Our results suggest that
minerals may be involved in neurodegenerative
processes.

Introduction
Spinal cord injury (SCI) induces devastating
neurological functional deficits. We hypothesize that
metal accumulation might be a trigger in injury to the
central nervous system. After injury, gray matter shows
less protection than white matter. We aim to quantify
metal repartition within the T9 and T10 segments of the
gray matter and to compare ipsi- and contralateral sides
to the hemi-sectioned spinal cord.

Results

Results (Cont.)

1)Ventral horn averages for Cu or Fe quantification (g/

cm 2) show important differences between the lesion
side and the control side of the spinal cord. These
differences also exist at distance to the lesion (caudal
T10).
Figure 1

Figure 3

NL-VH

LVH

xsem

xsem

% of
increase/de
crease
on lesion
side

Proximal to Lesion
T9
Distal to Lesion
T10

0.700.02

1.000.01

42%

0.350.01

0.390,01

11%

Proximal To Lesion
T9

0.130.002

0.120.003

-8%

0.100.002

0.120.004

20%

2
Mineral g/cm

Fe

Cu

Distal To Lesion
T10

Table 1 shows quantitative mineral repartitions within

the entire ventral horn and their the percentage of
variation induced by a chronic lesion to the spinal
cord.
2) Moreover local differences within the ventral horn
between ipsi- and contra-lateral side to the lesion
exist.
Figure 2
We suggest that some minerals such as Fe might not
affect less the distal neuronal population and are more
directly linked to the lesion insult. Therefore, these
minerals might not be involved in a cascade of events
inducing large and distant degenerative areas.

Mineral

g/cm2

Fe

Proximal To Lesion

NL-DVH

L-DVH

xsem

xsem

% of
increase/de
crease
on lesion
side

1.440.12

1.700.09

18%

0.450.05

0.450.00

0%

0.150.003

0.150.01

0%

0.110.002

0.130.004

18%

T9
Distal to Lesion
T10

Cu

Proximal to Lesion
T9
Distal to Lesion
T10

Table 2 shows quantitative data of minerals within the

dorsal part of the ventral horn. This suggests that
mineral affects differentially specific populations of
neurons such as here interneuronal population.

1)These results show that metal concentration induced

by chronic injury (2 weeks).
2) Our results show differential involvement of minerals
within the entire ventral horn after SCI.
3) The distance from the lesion is a factor involved in
mineral concentration
4) Local mineral accumulation in discrete areas of the
spinal cord correspond to anatomical and functional
specialization

Acknowledgements
This work was supported by NIH-EARDA grant 52457.
TP, SH were supported by NIH-MBRS-RISE R25
GM59218

Figure 4

Methods
Four male Sprague Dawley rats weighing 18010g were
housed at UIC animal facility. Animals were provided with
standard food and drinking water. The rats were
anesthetized using intraperitoneal injection of sodium
pentobarbital (60mg/kg ), (FisherScientific). Two
successive surgeries took place. During the first surgery a
spinal cord lesion was induced by a hemi-section at the T9
segment using a microsurgical scalpel and microscope.
The rats were allowed to recover from surgery and were
monitored for pain distress and sensory evaluation of the
lesion. Their weights were recorded every day for 3 days
and then every other day for the remainder of the 2-week
period. Thereafter, each rat was subjected to a second,
terminal surgical procedure. Their vertebral column was
exposed at the T10-T8 region and lamenectomies were
performed on vertebrae T8 to T10. T9-T10 segments of the
spinal cord were then excised and prepared for histological
sectioning (10m thickness) and X-Ray fluorescence
(XRF) analyses. The spinal cord samples lay on kapton
that was taped to a plastic holder. XRF set up and
scanning was conducted at the BioCat beam line at
Argonne National Laboratory. Samples were scanned on
the x-ray fluorescence microprobe using a 20m
continuous scan.

3) Differences between ipsi and contra-lateral sides to

the lesion are observed proximally to the lesion (T9) for
Fe and at a distance (T10) from the lesion for Cu. This
suggests that individual minerals are differentially
involved in neurodegenerative or regenerative (?)
cascades after SCI. Therefore some minerals might be
more involved in long axonal tracts survival (perhaps
Cu) than in local regulatory interneuronal networks
(perhaps Fe).

Conclusion