1. Maternal Insult and Embryonic Neurodevelopmental Damage
Hava Golan, Mahmoud Huleihel, Ira Kashtuzki-Philip, Maya Gerstein, Revital Mane, Miriam Stilman, Tamar Levav, Vered Kaplan
Dept of Developmental Genetics & Zlotowski center for Neuroscience
Energy failure
Transcriptional and translational regulation
Fetal hypoxia
Regeneration
Cell death
Immediate response
Delayed response

2. Study Design
Animals: Pregnant, Jackson Black C-57 mice at gestation day 17 were assigned to one of four groups:
Saline injections (i.p) in room air exposure (S+A),
Saline injections (i.p) with hypoxia chamber exposure (S+H),
Mg injections (i.p) in room air exposure (Mg+A),
Mg injections (i.p) with hypoxia chamber exposure (Mg+H).
Mg administration: 270 mg/kg followed by 27 mg/kg every 20 minutes for 4 hours. A second loading dose of 270 mg/kg was given at the end of the 4-hour period. Control mice were injected with saline following the same volume and schedule.
P 180
P 14
P 1
E17
E20
E18 P1 P7
P14
P21
P120-P180
4h pretreatment
Saline or Mg
2h hypoxia or air
9% O2
3% CO2

3. IS MAGNESIUM-SULFATE LOAD A PROPHLACTIC TREATMENT FOR PRENATAL HYPOXIA?
S+H Mg+H
Behavior
Motor impaired partial protection
Learning impaired short and long term Sort-term normal long-term not
Brain morphology
Cerebellum low PC density PC density normal
thinner granular layer not prevented
Cortex low cell density L 2-3,5-6 L5-6 protected
Hippocampus no effect thinner CA1 layer
GABAergic neurons
Cortex PV and CB decreased L2-3, 5-6 L2-3 partial protected
Hippocampus PV in CA3 decreased not prevented
Inhibitory synaptogenesis
Cortex not affected increased in all layers
Hippocapus VGAT in CA3 & hilus decreased not prevented
Glutamatergic synaptogenesis
Cerebellum reduced NR2AB density in PC at P1 normal NR2AB density
Cortex no effect no effect
Hippocampus reduced NR2AB in all regions at P1 normal NR2AB density

4. Behavior - Exploration in open field
First minute
Fifth minute
S+A
S+H
Mg+H

5. Motor Function Wire Clinging Footprint Pattern Vertical Pole*
Stride distance
(mm)
% of mice failed
% of Mice Stopped
Vertical Pole
% of mice failed
N=11-15 animals in each group
Golan et al., 2004

6. Morphology - Cerebellum
Cerebellum Clb 3
S+A
S+H
Mg+H * ** *
Purkinje cells density (cell/100mm)
Granular Layer Width (mm)
N=3-4 animals in each group, 3-4 section per animal
Golan et al., 2004

7. 2h 24h P-value < 0.01 P-value < 0.05 P-value < 0.05
SH - Control
MgH - Control
P-value < 0.01
634
291
196
SH - Control
MgH - Control 2h
1912
1108
594
P-value < 0.05
P-value < 0.05 57 48
250
SH - Control
MgH - Control
P-value < 0.01 9 12 66
SH - Control
MgH - Control
24h

8. Differential expression of genes
Number of genes
Time after hypoxia (hours)
Number of genes
folds change > 1.3, P<0.05

9. Spearman Rank Correlation
With centered data, Pairwise centroid linkage
Pattern 1
Pattern 2
Pattern 3

10. Pattern 1

11. Pattern 2

12. Pattern 3