1. DAPK1 Interaction with NMDA Receptor Mediates Brain Damage in Stroke
杨佳柠 临床医学（八年制）

2. Stroke [1] acute cerebrovascular diseases
brain disorders caused by pathologic processes involving blood vessels
3 main pathogenic mechanisms:
thrombotic occlusion
embolic occlusion
vascular rupture
ischemia
hemorrhage
[1]Robbins BASIC PATHOLOGY, 9th Edition

3. Keys DAPK1: Death-associated Protein Kinase 1 死亡相关蛋白激酶1
NMDAR: N-methyl-D- aspartate receptors N-甲基-D-天冬氨酸受体
Ischemic Brain Damage
Figure 1. Pierre Paoletti and Jacques Neyton. NMDA receptor subunits: function and pharmacology, Current Opinion in Pharmacology, Vol. 7:39–47, 2007

4. Methods & Results Any changes after neurons suffering from ischemia?
anti-NMDAR subunit NR2B antibodies
focal cerebral ischemia: middle cerebral artery occlusion (MCAO)
Ischemia recruits DAPK1 into NR2B complex.
Figure 2. Youming Lu, et al. DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke, Cell, Vol.140(2): , 2010

5. Methods & Results How does the DAPK1 interact with NR2B?
generating a series of NMDA receptor NR2B C-terminal deletion mutants
only the NR2B fragment capable of binding with DAPK1 (using a specific motif)
coexpressing cDAPK1 with the normal NR1/NR2B or the mutant NR1/NR2B S1303 A receptors (a Ser-1303 residue replaced with an Ala)
cDAPK1 increased the peak amplitude of the recombinant NR1/NR2B receptor currents, but not the mutant receptor

6. Methods & Results What happens after their interaction?
coexpressing the NR1/NR2B receptors with a constitutively active DAPK1 (cDAPK1), or the wild-type DAPK1 (wDAPK1)
DAPK1 activation increased the NR1/NR2B Ca2+ channel conductance
Figure 3. Robbins BASIC PATHOLOGY, 9th Edition

7. Methods & Results
Can neurons be protected if the interaction is inhibited?
DAPK1-/- mutant mice
Figure 4. Youming Lu, et al. DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke, Cell, Vol.140(2): , 2010

8. Methods & Results
Cortex
100μm
Striatum
CA1 A B
treated the DAPK1-/- and DAPK1+/+ mice with transient global ischemia (occlusion of the common carotid artery for 20 min), then reperfusion
6 days after, stained with Fluoro- Jade (FJ), a marker for degenerating neurons
Genetic deletion of DAPK1 protects neurons against Ischemic injury.
Figure 5. Youming Lu, et al. DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke, Cell, Vol.140(2): , 2010

9. Discussion
DAPK1 only mediating neuron’s pathologic function, without affecting the normal physiologic function
New targeting stroke therapy: DAPK1-NMDA receptor interaction
DAPK1-specific inhibitors or antagonists
Figure 6. Youming Lu, et al. DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke, Cell, Vol.140(2): , 2010