DAPKI Interaction with NMDA Receptor Mediates brain Damage in stroke 11307120230杨佳柠临床医学(八年制)
DAPK1 Interaction with NMDA Receptor Mediates Brain Damage in Stroke 11307120230 杨佳柠 临床医学（八年制）
Stroke o acute cerebrovascular diseases brain disorders caused by pathologic processes involving blood vessels 3 main pathogenic mechanisms: thrombotic occlusion embolic occlusion IschemIa vascUlar rupTure hemorrhage [] Robbins BASIC PATHOLOGY, 9h Edition
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
Keys NR1 NR2 Allosteric Site 2? DAPK1: Death-associatecdomains ifenprodil(NR2B) Protein Kinase 1 Site 3? 死亡相关蛋白激酶 Competitive agonists mains Glutamate NMDAR: N-methyl-D Site 4? aspartate receptors blockers N-甲基D-天冬氨酸受体 C-terminal Ischemic brain damage Current Opinion in Pharmacology Figure 1. Pierre Paolettiand Jacques Neyton. NMDA receptor subunits: function and pharmacology, Current Opinion in Phamacology, Vol 7: 39-47
Keys DAPK1: Death-associated Protein Kinase 1 死亡相关蛋白激酶1 NMDAR: N-methyl-Daspartate receptors N-甲基-D-天冬氨酸受体 Ischemic Brain Damage Figure 1. Pierre Paolettiand Jacques Neyton. NMDA receptor subunits: function and pharmacology, Current Opinion in Pharmacology, Vol. 7:39–47, 2007
Methods results A Any changes after neurons suffering from ischemia? Anti-NR2B NS IgG anti-NMDAR sUbunit NR2B -200 antibodies focal cerebral ischemia: middle 150 cerebral artery occlusion (MCAO Ischemia recruits daPkl into nr2b 45 complex. Figure 2. Youming Lu, et al. DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke, Cell, VoL. 140(2): 222-234, 2010
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):222-234, 2010
Methods results How does the daPI interact with nr2B2 generating a series of NMDA receptor NR2B C-terminal deletion mutants only the NR2B 292- 1304 fragment capable of binding with DAPKI(using a specific motif) coexpressing CDAPKI with the normal NRI/NR2B or the mutant NRI/NR2B $303 A receptors(a Ser-1303 residue replaced with an Ala) CDAPKI increased the peak amplitude of the recombinant NRT/NR2B receptor currents, but nof the mutant receptor
Methods & Results How does the DAPK1 interact with NR2B? generating a series of NMDA receptor NR2B C-terminal deletion mutants only the NR2B1292-1304 fragment capable of binding with DAPK1 (using a specific motif) coexpressingcDAPK1 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
Methods results Extracellular Injurious agen What happens after their int 00000n Mitochon Smooth ER coexpressing the NRI/NR Increased cytosolic Ca constitutively active DAPK Activation of DAPKI WDAPKI cellular enzymes D DAPKI activation Protease Endo- ATPase t mite nuclease transition channel conductance ↓ Phospho- Disruption DAMAGE DAMAGE ↓ATP Figure I-17 Sources and consequences of increased cytosolic calcium Figure 3. Robbins BASIC PATHOLOGY, 9th Edition in cell injury. ATP, adenosine triphosphate, ATPase, adenosine
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
Methods results Can neurons be protected if the interaction is inhibited? ATG st Stop BO B-geo Cort cA 500 50 P DAPKI-/-mutant mice Figure 4. Youming Lu, et al. DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke, Cell, VoL. 140(2): 222-234, 2010
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):222-234, 2010
Methods results Cortex Cortex treated the DAPKl-/-and DAPK1+/+ mice with transient global ischemia (occlusion of 100um 100um the common carotid artery for 20 min), then reperfusion Striatum Striatum. 6 days after, stained with Fluoro- Jade(FJ), a marker for degenerating neurons 100um 100um CA1 Ais* CA1 Genetic deletion of DAPKI protects neurons againsf Ischemic injury. 100 100um Figure 5. Youming Lu, et al. DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke, Cell, VoL. 140(2): 222-234, 2010
Methods & Results 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 FluoroJade (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):222-234, 2010 Cortex 100μm 100μm Cortex Striatum 100μm 100μm Striatum CA1 100μm 100μm CA1 A B
Discussion Stroke induction Stroke induction blocking peptide D DAPKl only mediating neuron’ s pathologic function, without affecting the normal NMD physiologic function Blocking peptide New targeting stroke therapy: DAPK1-NMDA Dcas receptor interaction DAPKI-specific inhibitors or Neuronal death Cell survival and antagonists Protection from stroke damage Figure 6. Youming Lu, et al. DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke, Cell, VoL. 140(2): 222-234, 2010
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):222-234, 2010