Regulation of NMDA receptors by serine proteases tissue plasminogen activator (tPA) and plasminogen plasmin

Title Page REGULATION OF NMDA RECEPTORS BY SERINE PROTEASES TISSUE PLASMINOGEN ACTIVATOR (tPA) AND PLASMINOGEN/PLASMIN NG KAY SIONG B. Appl. Sci. (Hons.), NUS A THESIS SUBMITTED FOR THE DEGREE OF DOCTOR OF PHILOSOPHY DEPARTMENT OF PHARMACOLOGY NATIONAL UNIVERSITY OF SINGAPORE 2011 Acknowledgements First and foremost, I would like to express my greatest gratitude to my supervisor, Dr. Low Chian Ming for giving me invaluable advice and guidance in the PhD project throughout the course of this postgraduate degree. I am deeply indebted to his time and patience and also the constant encouragement he has given me. I would also like to thank my co-supervisor, Prof. Peter Wong Tsun Hon, who has been providing assistance and advice whenever required. Thanks also go out to my fellow lab members, Ms. Cheong Yoke Ping and Ms. Zhang Yi Bin for their technical support; Ms. Karen Wee Siaw Ling and Ms. Leung How Wing for the constant intellectual discussion and encouragement. I am also very appreciative of past lab members Dr. Ng Fui Mee, Dr. Rema Vazhappilly, Dr Vivien Chow and Ms. Lim Peiqi, who shared their technical expertise and not forgetting Ms. Chen Jing Ting and Ms. Noella Anthony for their technical assistance. I would also like to express my appreciation to our collaborators, Prof. Stephen F. Traynelis (Emory University, Atlanta, GA) and Dr. Hiroyasu Furukawa (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY) for all the valuable advice. My sincere thanks also go to my thesis examiners for spending their precious time on my thesis. Special thanks also go out to my two mentors, Assoc. Prof. Tok Eng Soon and Dr. Alvin Teo for their support and assistance. Lastly, I would like to thank my family for their support and encouragement. The completion of this thesis would not have been possible without the support of many people. I sincerely thank all who have helped in making this postgraduate course a success. ii Table of Contents Title Page i Acknowledgements ii Table of Contents iii List of Publications vii Summary viii List of tables x List of figures xi Abbreviations xiii CHAPTER Introduction 1.1 Glutamate Receptors in the Mammalian Central Nervous System 1.2 Ionotropic Glutamate Receptors 1.3 NMDA Receptors in the Brain: Localization and Architecture 1.3.1 The NMDA receptor gene families and their localization 1.3.1.1 The NR1 subunit 1.3.1.2 The NR2 subunits 1.3.1.3 The NR3 subunits 1.3.2 Subunit topology 1.3.2.1 Amino-terminal domain (ATD) 1.3.2.2 S1S2 ligand-binding domain (LBD) 1.3.2.3 Transmembrane domain 1.3.2.4 Carboxyl terminal domain (CTD) 1.3.3 Structure of the NMDA receptor 4 8 10 11 11 12 1.4 NMDA Receptor Channel Properties and Pharmacology 1.4.1 Channel properties 1.4.1.1 NR2-containing NMDA receptors 1.4.1.2 NR3-containing NMDA receptors 1.4.2 Receptor pharmacology 1.4.2.1 Agonists 1.4.2.2 Competitive antagonists 1.4.2.3 Uncompetitive antagonists 13 14 14 15 15 15 16 18 1.5 NMDA Receptors at The Glutamatergic Synapse 1.5.1 Structure of the glutamatergic synapse 18 18 iii 1.5.2 NMDA receptors residing at the postsynaptic membrane 1.5.2.1 Subunit composition 1.5.2.2 Signaling mechanisms 21 21 22 1.6 Modulation of NMDA Receptors 1.6.1 Allosteric modulators 1.6.1.1 Protons 1.6.1.2 Zinc ions (Zn2+) 1.6.1.3 Polyamines 1.6.1.4 NR2B-selective allosteric antagonists 1.6.2 Functional regulation by phosphorylation 1.6.2.1 Regulation by serine/threonine phosphorylation 1.6.2.2 Regulation by tyrosine phosphorylation 1.6.3 Proteases as modulators 1.6.3.1 Matrix metalloproteinases (MMPs) 1.6.3.2 Calpain 1.6.3.3 Thrombin 1.6.3.4 Tissue-type plasminogen activator (tPA) 22 23 23 24 25 26 27 27 27 29 30 30 30 32 1.7 Tissue-type Plasminogen Activator (tPA) 1.7.1 tPA and stroke 1.7.2 tPA/plasminogen system in the brain 1.7.2.1 Expression 1.7.2.2 Regulation 32 32 33 34 34 1.8 tPA and the glutamatergic synapse 1.8.1 Structural modulation: Spine modelling 1.8.2 Molecular modulation: Synaptic plasticity 1.8.3 Molecular modulation: The NMDA receptor 1.8.3.1 tPA promotes neurotoxicity through the NMDA receptors 1.8.3.2 Plasmin cleavage of NMDA receptors 1.8.3.3 LRP and the NMDA receptor 1.8.3.4 tPA and NR2B-containing NMDA receptors 36 36 36 37 37 38 39 40 Thesis Objective 42 1.9 CHAPTER tPA-Induced Cleavage of the NR2B Subunits 43 2.1 Background and Objectives 44 2.2 Materials and Methods 44 2.3 Results tPA cleaves rat brain lysate NR2B subunit Antibody epitope mapping tPA cleaves the recombinant fusion protein MBP-ATD2B 51 51 52 58 2.4 Discussion and Conclusions 61 2.5 Conclusion 68 iv CHAPTER Plasmin Cleavage of NR1 and NR2B Subunits 69 3.1 Background and Objectives 70 3.2 Materials and Methods 70 3.3 Results Plasmin degrades NR2B Plasmin degrades NR1 tPA cleavage of NR2B is independent of plasmin 72 72 73 77 3.4 Discussion 79 3.5 Conclusions 81 CHAPTER Functional Consequence of Truncated NR2B-Containing NMDA Receptor 82 4.1 Background and Objectives 83 4.2 Materials and Methods 83 4.3 Results ATD-truncated NR2B forms functional NR1/NR2B-∆ATD-R67 receptors with reduced ifenprodil sensitivity Truncated NR2B reduces glycine potency Truncation of NR2B-ATD changes D-cycloserine efficacy and potency 87 87 89 89 4.4 Discussion 92 4.5 Conclusions 97 CHAPTER tPA-Induced Decrease of Synaptic NR2B Subunits 98 5.1 Background and Objectives 99 5.2 Materials and Methods 99 5.3 Results tPA decreases NR2B protein levels in the synaptic fraction tPA does not alter NR1 and NR2A protein levels 104 105 108 5.4 Discussion 108 5.5 Conclusions 116 CHAPTER Conclusion 118 6.1 Conclusion 119 6.2 Future Directions 124 v Reference 130 vi List of Publications 1) Wee XK, Ng KS, Leung HW, Cheong YP, Kong KH, Ng FM, Soh W, Lam Y, Low CM (2010) Mapping the high-affinity binding domain of 5-substituted benzimidazoles to the proximal N-terminus of the GluN2B subunit of the NMDA receptor. Br J Pharmacol 159:449-461. 2) Ng KS, Leung HW, Traynelis SF, Wong PTH, Furukawa H, Low CM. Ectodomain cleavage on the NR2B subunit by tissue plasminogen activator results in a functional truncated NMDA receptor with reduced ifenprodil and glycine affinities. (In preparation) Abstracts 1) Ng KS, Wong PTH, Low CM (2008) ‘Yin and Yang’ of FDA-approved clotbusting recombinant tissue plasminogen activator (tPA): Its proteolytic cleavage of NR2B subunit of NMDA receptor. (2nd Taiwan/Hong Kong(CU)/Singapore Meeting of Pharmacologists, Kaohsiung, Taiwan, Nov 2008 (Poster presentation)) 2) Low CM, Wee XK, Ng KA, Leung HW, Cheong YP, Kong KH, Ng FM, Soh WQ, Y Lam (2008) Benzimidazole derivatives bind at sub-nanomolar concentrations to recombinant protein of the NR2B amino-terminal domain of NMDA receptor. Soc Neurosci Abstr 131.4/D8. (38th Annual Meeting of Society for Neuroscience, Washington DC, USA 2008 (Poster presentation)) 3) Ng KS, Wong PTH, Low CM (2007) NR2B subunit of NMDA receptor is a new substrate for tissue plasminogen activator. Soc. Neurosci Abstr 678.17/F29. (37th Annual Meeting of Society for Neuroscience, San Diego, USA 2007 (Poster presentation)) 4) Ng KS, Traynelis SF, Wong PTH, Low CM (2007) Anti-Clotting Agent, Tissue Plasminogen Activator (tPA), cleaves NR2B Subunit of NMDA Receptor in Mammalian Brain. (Office of Life Sciences Conference, National University of Singapore, Singapore 2007 (Poster presentation)) vii Summary Tissue plasminogen activator (tPA) is an endogenous serine protease that is found in the vascular system and the central nervous system. The tPA/plasminogen proteolytic cascade which converts plasminogen to plasmin through tPA cleavage plays a critical role in dissolving blot clots and helps to maintain vascular patency. In the central nervous system, the tPA/plasminogen system is also involved in many processes ranging from synaptic plasticity to neurodegeneration. In particular, increasing evidence implicate tPA as an important neuromodulator of the N-methyl-Daspartate (NMDA) receptors. The aim of this thesis is to examine the modulation of NR2B-containing NMDA receptors by the tPA/plasminogen system. Through the analysis of tPA-treated rat brain lysates, I found that tPA can degrade the NR2B subunits of the NMDA receptors and this tPA-induced degradation was independent of plasmin. Peptide sequencing studies performed on the cleaved-off products obtained from the tPA treatment of a recombinant fusion protein containing the amino-terminal domain (ATD) of NR2B, revealed that tPA-mediated cleavage occurred at arginine 67 (Arg67) located in the ATD. Hence, I sought to examine how the deletion of a short peptide proximal to valine 68 (Val68) in the NR2B subunit, could alter NMDA receptor function. Electrophysiological studies on Xenopus laevis oocytes which heterologously expressed NR1 with the ATD-truncated form of NR2B (NR2B-ΔATD-R67) revealed a reduction in ifenprodil sensitivity. In addition, the potencies of glycine and Dcycloserine were reduced. Furthermore, the efficacy of D-cycloserine was enhanced when the amino acids 28-67 at the proximal end of the NR2B-ATD was deleted. Although the underlying mechanisms of the findings are unknown, these findings viii revealed that the amino acids proximal to Val68 could harbor critical determinants that could be important for the allosteric modulation of NMDA receptor channel properties. It is unknown whether putative tPA-induced NR2B-ATD cleavage of the NR2B or other forms of modulatory mechanisms of tPA on the NMDA receptors can change the NR2B-containing NMDA receptors levels in different subcellular compartments. This paradigm was examined through the acute tPA treatment of P14 whole hippocampi and subjecting treated-hippocampi to subcellular fractionation. The subsequent analysis of the different subcellular compartments revealed that tPA treatment led to a decrease in synaptic NR2B subunit levels in the hippocampus. In addition to examining the direct modulatory role of tPA on NR2B-containing NMDA receptors, the proteolytic effect of plasmin on NMDA receptors was also investigated. Both NR1 and NR2B were found to be proteolytic substrates of plasmin. My results demonstrated that the ATD, S2 and carboxyl-terminal domain (CTD) of NR1 may harbor potential plasmin cleavage sites, which are mostly consistent with the putative cleavage sites reported by other laboratories. In addition, I found that the NR2B subunit can be cleaved by plasmin at two potential sites residing in the CTD. New insights into the modulation of NR2B-containing NMDA receptors by the tPA/plasminogen system were presented in this thesis. Further studies into the underlying mechanisms engaged by tPA in the modulation of NMDA receptors would enable us to have a better understanding of the multi-faceted roles of tPA in the brain. (500 words) ix List of tables Table 1.1. Competitive NMDA receptor antagonists and their binding affinity to NR2 subunits 17 Table 1.2. Selected roles of proteases in the brain 29 Table 2.1. Alignment of critical residues around the scissile peptide bonds of tPA substrates. 63 Table 2.2. Concentrations of tPA used in various research reports 66 Table 5.1. NMDA receptor subunits protein levels after tPA (20 g/ml) treatment 112 x Gingrich MB, Traynelis SF (2000) Serine proteases and brain damage - is there a link? Trends Neurosci 23:399-407. Gingrich MB, Junge CE, Lyuboslavsky P, Traynelis SF (2000) Potentiation of NMDA receptor function by the serine protease thrombin. J Neurosci 20:4582-4595. Gloor S, Odink K, Guenther J, Nick H, Monard D (1986) A glia-derived neurite promoting factor with protease inhibitory activity belongs to the protease nexins. Cell 47:687-693. Goebel-Goody SM, Davies KD, Alvestad Linger RM, Freund RK, Browning MD (2009) Phospho-regulation of synaptic and extrasynaptic N-methyl-d-aspartate receptors in adult hippocampal slices. Neuroscience 158:1446-1459. Goldstein LB (2007) Acute Ischemic Stroke Treatment in 2007. Circulation 116:1504-1514. Goll DE, Thompson VF, Li H, Wei W, Cong J (2003) The calpain system. Physiol Rev 83:731-801. Gravanis I, Tsirka SE (2005) Tissue plasminogen activator and glial function. Glia 49:177-183. Groc L, Choquet D (2006) AMPA and NMDA glutamate receptor trafficking: multiple roads for reaching and leaving the synapse. Cell Tissue Res 326:423438. Groc L, Bard L, Choquet D (2009) Surface trafficking of N-methyl-d-aspartate receptors: Physiological and pathological perspectives. Neuroscience 158:4-18. Groc L, Choquet D, Stephenson FA, Verrier D, Manzoni OJ, Chavis P (2007) NMDA Receptor Surface Trafficking and Synaptic Subunit Composition Are Developmentally Regulated by the Extracellular Matrix Protein Reelin. J Neurosci 27:10165-10175. Groc L, Heine M, Cousins SL, Stephenson FA, Lounis B, Cognet L, Choquet D (2006) NMDA receptor surface mobility depends on NR2A-2B subunits. Proc Natl Acad Sci U S A 103:18769-18774. Gualandris A, Jones TE, Strickland S, Tsirka SE (1996) Membrane depolarization induces calcium-dependent secretion of tissue plasminogen activator. J Neurosci 16:2220-2225. Guttmann RP, Baker DL, Seifert KM, Cohen AS, Coulter DA, Lynch DR (2001) Specific proteolysis of the NR2 subunit at multiple sites by calpain. J Neurochem 78:1083-1093. Guttmann RP, Sokol S, Baker DL, Simpkins KL, Dong Y, Lynch DR (2002) Proteolysis of the N-Methyl-D-Aspartate Receptor by Calpain in Situ. J Pharmacol Exp Ther 302:1023-1030. Han X, Tomitori H, Mizuno S, Higashi K, Full C, Fukiwake T, Terui Y, Leewanich P, Nishimura K, Toida T, Williams K, Kashiwagi K, Igarashi K (2008) Binding of spermine and ifenprodil to a purified, soluble regulatory domain of the Nmethyl-D-aspartate receptor. J Neurochem 107:1566-1577. Hansen KB, Furukawa H, Traynelis SF (2010) Control of assembly and function of glutamate receptors by the amino-terminal domain. Mol Pharmacol. Hardingham GE (2009) Coupling of the NMDA receptor to neuroprotective and neurodestructive events. Biochem Soc Trans 37:1147-1160. Hardingham GE, Fukunaga Y, Bading H (2002) Extrasynaptic NMDARs oppose synaptic NMDARs by triggering CREB shut-off and cell death pathways. Nat Neurosci 5:405-414. 135 Hashimoto A, Oka T, Nishikawa T (1995) Extracellular concentration of endogenous free d-serine in the rat brain as revealed by in vivo microdialysis. Neuroscience 66:635-643. Hastings GA, Coleman TA, Haudenschild CC, Stefansson S, Smith EP, Barthlow R, Cherry S, Sandkvist M, Lawrence DA (1997) Neuroserpin, a brain-associated inhibitor of tissue plasminogen activator is localized primarily in neurons. Implications for the regulation of motor learning and neuronal survival. J Biol Chem 272:33062-33067. Herin GA, Aizenman E (2004) Amino terminal domain regulation of NMDA receptor function. Eur J Pharmacol 500:101-111. Herman Wolosker EDLBVNF (2008) d-Amino acids in the brain: d-serine in neurotransmission and neurodegeneration. FEBS Journal 275:3514-3526. Herz J, Strickland DK (2001) LRP: a multifunctional scavenger and signaling receptor. J Clin Invest 108:779-784. Hollmann M, Heinemann S (1994) Cloned Glutamate Receptors. Annual Review of Neuroscience 17:31-108. Hollmann M, Boulter J, Maron C, Beasley L, Sullivan J, Pecht G, Heinemann S (1993) Zinc potentiates agonist-induced currents at certain splice variants of the NMDA receptor. Neuron 10:943-954. Hood WF, Compton RP, Monahan JB (1989) D-cycloserine: a ligand for the Nmethyl-D-aspartate coupled glycine receptor has partial agonist characteristics. Neurosci Lett 98:91-95. Horak M, Wenthold RJ (2009) Different Roles of C-terminal Cassettes in the Trafficking of Full-length NR1 Subunits to the Cell Surface. Journal of Biological Chemistry 284:9683-9691. Hrafnkelsdottir T, Gudnason T, Wall U, Jern C, Jern S (2004) Regulation of local availability of active tissue-type plasminogen activator in vivo in man. J Thromb Haemost 2:1960-1968. Huang YY, Bach ME, Lipp HP, Zhuo M, Wolfer DP, Hawkins RD, Schoonjans L, Kandel ER, Godfraind JM, Mulligan R, Collen D, Carmeliet P (1996) Mice lacking the gene encoding tissue-type plasminogen activator show a selective interference with late-phase long-term potentiation in both Schaffer collateral and mossy fiber pathways. Proc Natl Acad Sci U S A 93:8699-8704. Huttner WB, Schiebler W, Greengard P, De Camilli P (1983) Synapsin I (protein I), a nerve terminal-specific phosphoprotein. III. Its association with synaptic vesicles studied in a highly purified synaptic vesicle preparation. J Cell Biol 96:1374-1388. Ikeda K, Nagasawa M, Mori H, Araki K, Sakimura K, Watanabe M, Inoue Y, Mishina M (1992) Cloning and expression of the [var epsilon]4 subunit of the NMDA receptor channel. FEBS Letters 313:34-38. Jin R, Singh SK, Gu S, Furukawa H, Sobolevsky AI, Zhou J, Jin Y, Gouaux E (2009) Crystal structure and association behaviour of the GluR2 amino-terminal domain. EMBO J 28:1812-1823. Johnson JW, Ascher P (1987) Glycine potentiates the NMDA response in cultured mouse brain neurons. Nature 325:529-531. Jones KS, VanDongen HMA, VanDongen AMJ (2002) The NMDA Receptor M3 Segment Is a Conserved Transduction Element Coupling Ligand Binding to Channel Opening. J Neurosci 22:2044-2053. 136 Karakas E, Simorowski N, Furukawa H (2009) Structure of the zinc-bound aminoterminal domain of the NMDA receptor NR2B subunit. EMBO J 28:39103920. Kassam G, Le BH, Choi KS, Kang HM, Fitzpatrick SL, Louie P, Waisman DM (1998) The p11 subunit of the annexin II tetramer plays a key role in the stimulation of t-PA-dependent plasminogen activation. Biochemistry 37:16958-16966. Kennedy MJ, Ehlers MD (2006) Organelles and trafficking machinery for postsynaptic plasticity. Annu Rev Neurosci 29:325-362. Kenny AV, Cousins SL, Pinho L, Stephenson FA (2009) The Integrity of the Glycine Co-agonist Binding Site of N-Methyl-d-aspartate Receptors Is a Functional Quality Control Checkpoint for Cell Surface Delivery. Journal of Biological Chemistry 284:324-333. Kew JNC, Kemp JA (2005) Ionotropic and metabotropic glutamate receptor structure and pharmacology. Psychopharmacology 179:4-29. Kew JNC, Richards JG, Mutel V, Kemp JA (1998) Developmental Changes in NMDA Receptor Glycine Affinity and Ifenprodil Sensitivity Reveal Three Distinct Populations of NMDA Receptors in Individual Rat Cortical Neurons. J Neurosci 18:1935-1943. Khalilov I, Esclapez M, Medina I, Aggoun D, Lamsa K, Leinekugel X, Khazipov R, Ben-Ari Y (1997) A novel in vitro preparation: the intact hippocampal formation. Neuron 19:743-749. Kim E, Sheng M (2004) PDZ domain proteins of synapses. Nat Rev Neurosci 5:771781. Kleckner N, Dingledine R (1988) Requirement for glycine in activation of NMDAreceptors expressed in Xenopus oocytes. Science 241:835-837. Klein M, Pieri I, Uhlmann F, Pfizenmaier K, Eisel U (1998) Cloning and characterization of promoter and 5'-UTR of the NMDA receptor subunit [epsilon]2: evidence for alternative splicing of 5'-non-coding exon. Gene 208:259-269. Knecht W, Cottrell GS, Amadesi S, Mohlin J, Skaregarde A, Gedda K, Peterson A, Chapman K, Hollenberg MD, Vergnolle N, Bunnett NW (2007) Trypsin IV or mesotrypsin and p23 cleave protease-activated receptors and to induce inflammation and hyperalgesia. J Biol Chem 282:26089-26100. Köhr G (2006) NMDA receptor function: subunit composition versus spatial distribution. Cell and Tissue Research 326:439-446. Kornau HC (2009) Postsynaptic Density/Architecture at Excitatory Synapses. In: Encyclopedia of Neuroscience (Larry RS, ed), pp 809-815. Oxford: Academic Press. Krapivinsky G, Krapivinsky L, Manasian Y, Ivanov A, Tyzio R, Pellegrino C, BenAri Y, Clapham DE, Medina I (2003) The NMDA receptor is coupled to the ERK pathway by a direct interaction between NR2B and RasGRF1. Neuron 40:775-784. Kumar J, Schuck P, Jin R, Mayer ML (2009) The N-terminal domain of GluR6subtype glutamate receptor ion channels. Nat Struct Mol Biol 16:631-638. Kuner T, Schoepfer R (1996) Multiple Structural Elements Determine Subunit Specificity of Mg2+ Block in NMDA Receptor Channels. J Neurosci 16:35493558. Kuryatov A, Laube B, Betz H, Kuhse J (1994) Mutational analysis of the glycinebinding site of the NMDA receptor: Structural similarity with bacterial amino acid-binding proteins. 12:1291-1300. 137 Kussius CL, Popescu GK (2009) Kinetic basis of partial agonism at NMDA receptors. Nat Neurosci 12:1114-1120. Kutsuwada T, Kashiwabuchi N, Mori H, Sakimura K, Kushiya E, Araki K, Meguro H, Masaki H, Kumanishi T, Arakawa M, et al. (1992) Molecular diversity of the NMDA receptor channel. Nature 358:36-41. Kvajo M, Albrecht H, Meins M, Hengst U, Troncoso E, Lefort S, Kiss JZ, Petersen CC, Monard D (2004) Regulation of brain proteolytic activity is necessary for the in vivo function of NMDA receptors. J Neurosci 24:9734-9743. LacKamp A, Zhang GC, Mao LM, Fibuch EE, Wang JQ (2009) Loss of surface Nmethyl-D-aspartate receptor proteins in mouse cortical neurones during anaesthesia induced by chloral hydrate in vivo. Br J Anaesth 102:515-522. Lan JY, Skeberdis VA, Jover T, Grooms SY, Lin Y, Araneda RC, Zheng X, Bennett MV, Zukin RS (2001) Protein kinase C modulates NMDA receptor trafficking and gating. Nat Neurosci 4:382-390. Lau CG, Zukin RS (2007) NMDA receptor trafficking in synaptic plasticity and neuropsychiatric disorders. Nat Rev Neurosci 8:413-426. Laube B, Kuhse J, Betz H (1998) Evidence for a Tetrameric Structure of Recombinant NMDA Receptors. J Neurosci 18:2954-2961. Laurie D, Seeburg P (1994a) Regional and developmental heterogeneity in splicing of the rat brain NMDAR1 mRNA. J Neurosci 14:3180-3194. Laurie DJ, Seeburg PH (1994b) Ligand affinities at recombinant N-methyl-Daspartate receptors depend on subunit composition. Eur J Pharmacol 268:335345. Lavezzari G, McCallum J, Lee R, Roche KW (2003) Differential binding of the AP-2 adaptor complex and PSD-95 to the C-terminus of the NMDA receptor subunit NR2B regulates surface expression. Neuropharmacology 45:729-737. Lavezzari G, McCallum J, Dewey CM, Roche KW (2004) Subunit-Specific Regulation of NMDA Receptor Endocytosis. J Neurosci 24:6383-6391. Lee CJ, Mannaioni G, Yuan H, Woo DH, Gingrich MB, Traynelis SF (2007) Astrocytic control of synaptic NMDA receptors. J Physiol 581:1057-1081. Lee H-K (2006) Synaptic plasticity and phosphorylation. Pharmacology & Therapeutics 112:810-832. Legendre P, Westbrook GL (1991) Ifenprodil blocks N-methyl-D-aspartate receptors by a two-component mechanism. Mol Pharmacol 40:289-298. Leveille F, El Gaamouch F, Gouix E, Lecocq M, Lobner D, Nicole O, Buisson A (2008) Neuronal viability is controlled by a functional relation between synaptic and extrasynaptic NMDA receptors. FASEB J 22:4258-4271. Li J, Shen H, Naus CC, Zhang L, Carlen PL (2001) Upregulation of gap junction connexin 32 with epileptiform activity in the isolated mouse hippocampus. Neuroscience 105:589-598. Liberatore GT, Samson A, Bladin C, Schleuning WD, Medcalf RL (2003) Vampire bat salivary plasminogen activator (desmoteplase): a unique fibrinolytic enzyme that does not promote neurodegeneration. Stroke 34:537-543. Lin Y, Jover-Mengual T, Wong J, Bennett MVL, Zukin RS (2006) PSD-95 and PKC converge in regulating NMDA receptor trafficking and gating. Proceedings of the National Academy of Sciences 103:19902-19907. Liu D, Cheng T, Guo H, Fernandez JA, Griffin JH, Song X, Zlokovic BV (2004a) Tissue plasminogen activator neurovascular toxicity is controlled by activated protein C. Nat Med 10:1379-1383. 138 Liu XB, Murray KD, Jones EG (2004b) Switching of NMDA receptor 2A and 2B subunits at thalamic and cortical synapses during early postnatal development. J Neurosci 24:8885-8895. Lochner JE, Honigman LS, Grant WF, Gessford SK, Hansen AB, Silverman MA, Scalettar BA (2006) Activity-dependent release of tissue plasminogen activator from the dendritic spines of hippocampal neurons revealed by livecell imaging. J Neurobiol 66:564-577. Lodge D (2009) The history of the pharmacology and cloning of ionotropic glutamate receptors and the development of idiosyncratic nomenclature. Neuropharmacology 56:6-21. Low C-M, Zheng F, Lyuboslavsky P, Traynelis SF (2000) Molecular determinants of coordinated proton and zinc inhibition of N-methyl-d-aspartate NR1/NR2A receptors. Proceedings of the National Academy of Sciences of the United States of America 97:11062-11067. Low CM, Lyuboslavsky P, French A, Le P, Wyatte K, Thiel WH, Marchan EM, Igarashi K, Kashiwagi K, Gernert K, Williams K, Traynelis SF, Zheng F (2003) Molecular determinants of proton-sensitive N-methyl-D-aspartate receptor gating. Mol Pharmacol 63:1212-1222. Lu W, Man H, Ju W, Trimble WS, MacDonald JF, Wang YT (2001) Activation of synaptic NMDA receptors induces membrane insertion of new AMPA receptors and LTP in cultured hippocampal neurons. Neuron 29:243-254. Lu WY, Xiong ZG, Lei S, Orser BA, Dudek E, Browning MD, MacDonald JF (1999) G-protein-coupled receptors act via protein kinase C and Src to regulate NMDA receptors. Nat Neurosci 2:331-338. Madani R, Hulo S, Toni N, Madani H, Steimer T, Muller D, Vassalli JD (1999) Enhanced hippocampal long-term potentiation and learning by increased neuronal expression of tissue-type plasminogen activator in transgenic mice. EMBO J 18:3007-3012. Madry C, Mesic I, Betz H, Laube B (2007a) The N-Terminal Domains of both NR1 and NR2 Subunits Determine Allosteric Zn2+ Inhibition and Glycine Affinity of N-Methyl-D-aspartate Receptors. Mol Pharmacol 72:1535-1544. Madry C, Mesic I, Bartholomäus I, Nicke A, Betz H, Laube B (2007b) Principal role of NR3 subunits in NR1/NR3 excitatory glycine receptor function. Biochemical and Biophysical Research Communications 354:102-108. Makarova A, Mikhailenko I, Bugge TH, List K, Lawrence DA, Strickland DK (2003) The low density lipoprotein receptor-related protein modulates protease activity in the brain by mediating the cellular internalization of both neuroserpin and neuroserpin-tissue-type plasminogen activator complexes. J Biol Chem 278:50250-50258. Mannaioni G, Orr AG, Hamill CE, Yuan H, Pedone KH, McCoy KL, Palmini RB, Junge CE, Lee CJ, Yepes M, Hepler JR, Traynelis SF (2008) Plasmin Potentiates Synaptic N-Methyl-D-aspartate Receptor Function in Hippocampal Neurons through Activation of Protease-activated Receptor-1. J Biol Chem 283:20600-20611. Martin AM, Kuhlmann C, Trossbach S, Jaeger S, Waldron E, Roebroek A, Luhmann HJ, Laatsch A, Weggen S, Lessmann V, Pietrzik CU (2008) The functional role of the second NPXY motif of the LRP1 beta-chain in tissue-type plasminogen activator-mediated activation of N-methyl-D-aspartate receptors. J Biol Chem 283:12004-12013. 139 Masuko T, Kashiwagi K, Kuno T, Nguyen ND, Pahk AJ, Fukuchi J, Igarashi K, Williams K (1999) A regulatory domain (R1-R2) in the amino terminus of the N-methyl-D-aspartate receptor: effects of spermine, protons, and ifenprodil, and structural similarity to bacterial leucine/isoleucine/valine binding protein. Mol Pharmacol 55:957-969. Mataga N, Mizuguchi Y, Hensch TK (2004) Experience-dependent pruning of dendritic spines in visual cortex by tissue plasminogen activator. Neuron 44:1031-1041. Matsuda K, Kamiya Y, Matsuda S, Yuzaki M (2002) Cloning and characterization of a novel NMDA receptor subunit NR3B: a dominant subunit that reduces calcium permeability. Brain Res Mol Brain Res 100:43-52. Matsuda K, Fletcher M, Kamiya Y, Yuzaki M (2003) Specific Assembly with the NMDA Receptor 3B Subunit Controls Surface Expression and Calcium Permeability of NMDA Receptors. J Neurosci 23:10064-10073. Matsui T, Sekiguchi M, Hashimoto A, Tomita U, Nishikawa T, Wada K (1995) Functional comparison of D-serine and glycine in rodents: the effect on cloned NMDA receptors and the extracellular concentration. J Neurochem 65:454458. Matys T, Strickland S (2003) Tissue plasminogen activator and NMDA receptor cleavage. Nat Med 9:371-372; author reply 372-373. May P, Rohlmann A, Bock HH, Zurhove K, Marth JD, Schomburg ED, Noebels JL, Beffert U, Sweatt JD, Weeber EJ, Herz J (2004) Neuronal LRP1 functionally associates with postsynaptic proteins and is required for normal motor function in mice. Mol Cell Biol 24:8872-8883. Mayer ML, Westbrook GL, Guthrie PB (1984) Voltage-dependent block by Mg2+ of NMDA responses in spinal cord neurones. Nature 309:261-263. McBain CJ, Mayer ML (1994) N-methyl-D-aspartic acid receptor structure and function. Physiol Rev 74:723-760. McBain CJ, Kleckner NW, Wyrick S, Dingledine R (1989) Structural requirements for activation of the glycine coagonist site of N-methyl-D-aspartate receptors expressed in Xenopus oocytes. Mol Pharmacol 36:556-565. McGurk JF, Bennett MV, Zukin RS (1990) Polyamines potentiate responses of Nmethyl-D-aspartate receptors expressed in xenopus oocytes. Proc Natl Acad Sci U S A 87:9971-9974. Medina MG, Ledesma MD, Dominguez JE, Medina M, Zafra D, Alameda F, Dotti CG, Navarro P (2005) Tissue plasminogen activator mediates amyloidinduced neurotoxicity via Erk1/2 activation. EMBO J 24:1706-1716. Melchor JP, Strickland S (2005) Tissue plasminogen activator in central nervous system physiology and pathology. Thromb Haemost 93:655-660. Meyers PM, Schumacher HC, Higashida RT, Barnwell SL, Creager MA, Gupta R, McDougall CG, Pandey DK, Sacks D, Wechsler LR (2009) Indications for the Performance of Intracranial Endovascular Neurointerventional Procedures: A Scientific Statement From the American Heart Association Council on Cardiovascular Radiology and Intervention, Stroke Council, Council on Cardiovascular Surgery and Anesthesia, Interdisciplinary Council on Peripheral Vascular Disease, and Interdisciplinary Council on Quality of Care and Outcomes Research. Circulation 119:2235-2249. Michaluk P, Mikasova L, Groc L, Frischknecht R, Choquet D, Kaczmarek L (2009) Matrix metalloproteinase-9 controls NMDA receptor surface diffusion through integrin beta1 signaling. J Neurosci 29:6007-6012. 140 Mony L, Kew JN, Gunthorpe MJ, Paoletti P (2009a) Allosteric modulators of NR2Bcontaining NMDA receptors: molecular mechanisms and therapeutic potential. Br J Pharmacol 157:1301-1317. Mony L, Krzaczkowski L, Leonetti M, Le Goff A, Alarcon K, Neyton J, Bertrand HO, Acher F, Paoletti P (2009b) Structural basis of NR2B-selective antagonist recognition by N-methyl-D-aspartate receptors. Mol Pharmacol 75:60-74. Monyer H, Burnashev N, Laurie DJ, Sakmann B, Seeburg PH (1994) Developmental and regional expression in the rat brain and functional properties of four NMDA receptors. 12:529-540. Mori H, Mishina M (1995) Structure and function of the NMDA receptor channel. Neuropharmacology 34:1219-1237. Mothet J-P, Parent AT, Wolosker H, Brady RO, Linden DJ, Ferris CD, Rogawski MA, Snyder SH (2000) d-Serine is an endogenous ligand for the glycine site of the N-methyl-d-aspartate receptor. Proceedings of the National Academy of Sciences of the United States of America 97:4926-4931. Mott DD, Doherty JJ, Zhang S, Washburn MS, Fendley MJ, Lyuboslavsky P, Traynelis SF, Dingledine R (1998) Phenylethanolamines inhibit NMDA receptors by enhancing proton inhibition. Nat Neurosci 1:659-667. Mott JD, Werb Z (2004) Regulation of matrix biology by matrix metalloproteinases. Curr Opin Cell Biol 16:558-564. Nadif Kasri N, Nakano-Kobayashi A, Malinow R, Li B, Van Aelst L (2009) The Rholinked mental retardation protein oligophrenin-1 controls synapse maturation and plasticity by stabilizing AMPA receptors. Genes Dev 23:1289-1302. Nagappan G, Zaitsev E, Senatorov VV, Jr., Yang J, Hempstead BL, Lu B (2009) Control of extracellular cleavage of ProBDNF by high frequency neuronal activity. Proc Natl Acad Sci U S A 106:1267-1272. Nagy Z, Kolev K, Csonka E, Vastag M, Machovich R (1998) Perturbation of the integrity of the blood-brain barrier by fibrinolytic enzymes. Blood Coagul Fibrinolysis 9:471-478. Nakagami Y, Abe K, Nishiyama N, Matsuki N (2000) Laminin degradation by plasmin regulates long-term potentiation. J Neurosci 20:2003-2010. Nakanishi N, Tu S, Shin Y, Cui J, Kurokawa T, Zhang D, Chen H-SV, Tong G, Lipton SA (2009) Neuroprotection by the NR3A Subunit of the NMDA Receptor. J Neurosci 29:5260-5265. Nakazawa T, Komai S, Tezuka T, Hisatsune C, Umemori H, Semba K, Mishina M, Manabe T, Yamamoto T (2001) Characterization of Fyn-mediated Tyrosine Phosphorylation Sites on GluR (NR2B) Subunit of theN-Methyl-d-aspartate Receptor. Journal of Biological Chemistry 276:693-699. Newpher TM, Ehlers MD (2008) Glutamate receptor dynamics in dendritic microdomains. Neuron 58:472-497. Neyton J, Paoletti P (2006) Relating NMDA Receptor Function to Receptor Subunit Composition: Limitations of the Pharmacological Approach. J Neurosci 26:1331-1333. Ng F-M, Geballe M, Snyder J, Traynelis S, Low C-M (2008) Structural insights into phenylethanolamines high-affinity binding site in NR2B from binding and molecular modeling studies. Molecular Brain 1:16. Ng FM, Soh W, Geballe MT, Low CM (2007) Improving solubility of NR2B aminoterminal domain of N-methyl-d-aspartate receptor expressed in Escherichia coli. Biochem Biophys Res Commun 362:69-74. 141 Nicole O, Docagne F, Ali C, Margaill I, Carmeliet P, MacKenzie ET, Vivien D, Buisson A (2001) The proteolytic activity of tissue-plasminogen activator enhances NMDA receptor-mediated signaling. Nat Med 7:59-64. Nishi M, Hinds H, Lu H-P, Kawata M, Hayashi Y (2001) Motoneuron-Specific Expression of NR3B, a Novel NMDA-Type Glutamate Receptor Subunit That Works in a Dominant-Negative Manner. J Neurosci 21:185RC-. Niswender CM, Conn PJ (2010) Metabotropic Glutamate Receptors: Physiology, Pharmacology, and Disease. Annual Review of Pharmacology and Toxicology 50:295-322. Norris EH, Strickland S (2007) Modulation of NR2B-regulated contextual fear in the hippocampus by the tissue plasminogen activator system. Proc Natl Acad Sci U S A 104:13473-13478. Okabe S (2007) Molecular anatomy of the postsynaptic density. Mol Cell Neurosci 34:503-518. Overall CM, Blobel CP (2007) In search of partners: linking extracellular proteases to substrates. Nat Rev Mol Cell Biol 8:245-257. Pahk AJ, Williams K (1997) Influence of extracellular pH on inhibition by ifenprodil at N-methyl-D-aspartate receptors in Xenopus oocytes. Neurosci Lett 225:2932. Pang PT, Lu B (2004) Regulation of late-phase LTP and long-term memory in normal and aging hippocampus: role of secreted proteins tPA and BDNF. Ageing Res Rev 3:407-430. Pang PT, Teng HK, Zaitsev E, Woo NT, Sakata K, Zhen S, Teng KK, Yung WH, Hempstead BL, Lu B (2004) Cleavage of proBDNF by tPA/plasmin is essential for long-term hippocampal plasticity. Science 306:487-491. Paoletti P, Neyton J (2007) NMDA receptor subunits: function and pharmacology. Curr Opin Pharmacol 7:39-47. Paoletti P, Ascher P, Neyton J (1997) High-Affinity Zinc Inhibition of NMDA NR1NR2A Receptors. J Neurosci 17:5711-5725. Paoletti P, Vergnano AM, Barbour B, Casado M (2009) Zinc at glutamatergic synapses. Neuroscience 158:126-136. Paoletti P, Perin-Dureau F, Fayyazuddin A, Le Goff A, Callebaut I, Neyton J (2000) Molecular organization of a zinc binding n-terminal modulatory domain in a NMDA receptor subunit. Neuron 28:911-925. Papadakis M, Hawkins LM, Stephenson FA (2004) Appropriate NR1-NR1 Disulfidelinked Homodimer Formation Is Requisite for Efficient Expression of Functional, Cell Surface N-Methyl-D-aspartate NR1/NR2 Receptors. Journal of Biological Chemistry 279:14703-14712. Papadia S, Soriano FX, Leveille F, Martel M-A, Dakin KA, Hansen HH, Kaindl A, Sifringer M, Fowler J, Stefovska V, McKenzie G, Craigon M, Corriveau R, Ghazal P, Horsburgh K, Yankner BA, Wyllie DJA, Ikonomidou C, Hardingham GE (2008) Synaptic NMDA receptor activity boosts intrinsic antioxidant defenses. Nat Neurosci 11:476-487. Park L, Gallo EF, Anrather J, Wang G, Norris EH, Paul J, Strickland S, Iadecola C (2008) Key role of tissue plasminogen activator in neurovascular coupling. Proc Natl Acad Sci U S A 105:1073-1078. Parmer RJ, Mahata M, Mahata S, Sebald MT, O'Connor DT, Miles LA (1997) Tissue plasminogen activator (t-PA) is targeted to the regulated secretory pathway. Catecholamine storage vesicles as a reservoir for the rapid release of t-PA. J Biol Chem 272:1976-1982. 142 Patneau D, Mayer M (1990) Structure-activity relationships for amino acid transmitter candidates acting at N-methyl-D-aspartate and quisqualate receptors. J Neurosci 10:2385-2399. Pauly T, Ratliff M, Pietrowski E, Neugebauer R, Schlicksupp A, Kirsch J, Kuhse J (2008) Activity-dependent shedding of the NMDA receptor glycine binding site by matrix metalloproteinase 3: a PUTATIVE mechanism of postsynaptic plasticity. PLoS ONE 3:e2681. Pawlak R, Magarinos AM, Melchor J, McEwen B, Strickland S (2003) Tissue plasminogen activator in the amygdala is critical for stress-induced anxietylike behavior. Nat Neurosci 6:168-174. Pawlak R, Melchor JP, Matys T, Skrzypiec AE, Strickland S (2005a) Ethanolwithdrawal seizures are controlled by tissue plasminogen activator via modulation of NR2B-containing NMDA receptors. Proc Natl Acad Sci U S A 102:443-448. Pawlak R, Rao BS, Melchor JP, Chattarji S, McEwen B, Strickland S (2005b) Tissue plasminogen activator and plasminogen mediate stress-induced decline of neuronal and cognitive functions in the mouse hippocampus. Proc Natl Acad Sci U S A 102:18201-18206. Pawlak R, Nagai N, Urano T, Napiorkowska-Pawlak D, Ihara H, Takada Y, Collen D, Takada A (2002) Rapid, specific and active site-catalyzed effect of tissueplasminogen activator on hippocampus-dependent learning in mice. Neuroscience 113:995-1001. Perez-Otano I, Schulteis CT, Contractor A, Lipton SA, Trimmer JS, Sucher NJ, Heinemann SF (2001) Assembly with the NR1 Subunit Is Required for Surface Expression of NR3A-Containing NMDA Receptors. J Neurosci 21:1228-1237. Perin-Dureau F, Rachline J, Neyton J, Paoletti P (2002) Mapping the binding site of the neuroprotectant ifenprodil on NMDA receptors. J Neurosci 22:5955-5965. Perrin BJ, Huttenlocher A (2002) Calpain. Int J Biochem Cell Biol 34:722-725. Petralia RS, Wang YX, Hua F, Yi Z, Zhou A, Ge L, Stephenson FA, Wenthold RJ (2010) Organization of NMDA receptors at extrasynaptic locations. Neuroscience 167:68-87. Pinheiro PS, Mulle C (2008) Presynaptic glutamate receptors: physiological functions and mechanisms of action. Nat Rev Neurosci 9:423-436. Polavarapu R, Gongora MC, Yi H, Ranganthan S, Lawrence DA, Strickland D, Yepes M (2007) Tissue-type plasminogen activator-mediated shedding of astrocytic low-density lipoprotein receptor-related protein increases the permeability of the neurovascular unit. Blood 109:3270-3278. Popescu G, Robert A, Howe JR, Auerbach A (2004) Reaction mechanism determines NMDA receptor response to repetitive stimulation. Nature 430:790-793. Prybylowski K, Chang K, Sans N, Kan L, Vicini S, Wenthold RJ (2005) The Synaptic Localization of NR2B-Containing NMDA Receptors Is Controlled by Interactions with PDZ Proteins and AP-2. Neuron 47:845-857. Qian A, Buller AL, Johnson JW (2005) NR2 subunit-dependence of NMDA receptor channel block by external Mg2+. The Journal of Physiology 562:319-331. Qian Z, Gilbert ME, Colicos MA, Kandel ER, Kuhl D (1993) Tissue-plasminogen activator is induced as an immediate-early gene during seizure, kindling and long-term potentiation. Nature 361:453-457. Qiu S, Zhang X-m, Cao J-y, Yang W, Yan Y-g, Shan L, Zheng J, Luo J-h (2009) An Endoplasmic Reticulum Retention Signal Located in the Extracellular Amino- 143 terminal Domain of the NR2A Subunit of N-Methyl-d-aspartate Receptors. Journal of Biological Chemistry 284:20285-20298. Rachline J, Perin-Dureau F, Le Goff A, Neyton J, Paoletti P (2005) The Micromolar Zinc-Binding Domain on the NMDA Receptor Subunit NR2B. J Neurosci 25:308-317. Rafiki A, Bernard A, Medina I, Gozlan H, Khrestchatisky M (2000) Characterization in Cultured Cerebellar Granule Cells and in the Developing Rat Brain of mRNA Variants for the NMDA Receptor 2C Subunit. Journal of Neurochemistry 74:1798-1808. Rambhadran A, Gonzalez J, Jayaraman V (2010) Subunit arrangement in N-methylD-aspartate (NMDA) receptors. J Biol Chem 285:15296-15301. Reddrop C, Moldrich RX, Beart PM, Farso M, Liberatore GT, Howells DW, Petersen KU, Schleuning WD, Medcalf RL (2005) Vampire bat salivary plasminogen activator (desmoteplase) inhibits tissue-type plasminogen activator-induced potentiation of excitotoxic injury. Stroke 36:1241-1246. Roche KW, Raymond LA, Blackstone C, Huganir RL (1994) Transmembrane topology of the glutamate receptor subunit GluR6. Journal of Biological Chemistry 269:11679-11682. Roche KW, Standley S, McCallum J, Dune Ly C, Ehlers MD, Wenthold RJ (2001) Molecular determinants of NMDA receptor internalization. Nat Neurosci 4:794-802. Rock DM, MacDonald RL (1992) Spermine and related polyamines produce a voltage-dependent reduction of N-methyl-D-aspartate receptor single-channel conductance. Molecular Pharmacology 42:157-164. Rock DM, Macdonald RL (1995) Polyamine regulation of N-methyl-D-aspartate receptor channels. Annu Rev Pharmacol Toxicol 35:463-482. Rosahl TW, Wingrove PB, Hunt V, Fradley RL, Lawrence JMK, Heavens RP, Treacey P, Usala M, Macaulay A, Bonnert TP, Whiting PJ, Wafford KA (2006) A genetically modified mouse model probing the selective action of ifenprodil at the N-methyl-d-aspartate type 2B receptor. Molecular and Cellular Neuroscience 33:47-56. Rosenberg GA (2009) Matrix metalloproteinases and their multiple roles in neurodegenerative diseases. Lancet Neurol 8:205-216. Ryan T, Emes R, Grant S, Komiyama N (2008) Evolution of NMDA receptor cytoplasmic interaction domains: implications for organisation of synaptic signalling complexes. BMC Neuroscience 9:6. Salles FJ, Strickland S (2002) Localization and regulation of the tissue plasminogen activator-plasmin system in the hippocampus. J Neurosci 22:2125-2134. Salter MW, Kalia LV (2004) Src kinases: a hub for NMDA receptor regulation. Nat Rev Neurosci 5:317-328. Samson AL, Medcalf RL (2006) Tissue-type plasminogen activator: a multifaceted modulator of neurotransmission and synaptic plasticity. Neuron 50:673-678. Samson AL, Nevin ST, Croucher D, Niego B, Daniel PB, Weiss TW, Moreno E, Monard D, Lawrence DA, Medcalf RL (2008) Tissue-type plasminogen activator requires a co-receptor to enhance NMDA receptor function. J Neurochem 107:1091-1101. Sans N, Racca C, Petralia RS, Wang YX, McCallum J, Wenthold RJ (2001) Synapseassociated protein 97 selectively associates with a subset of AMPA receptors early in their biosynthetic pathway. J Neurosci 21:7506-7516. 144 Sappino A-P, Madani R, Huarte J, Belin D, Kiss JZ, Wohlwend A, Vassalli J-D (1993) Extracellular proteolysis in the adult murine brain. J Clin Invest 92:679-685. Schell MJ, Molliver ME, Snyder SH (1995) D-serine, an endogenous synaptic modulator: localization to astrocytes and glutamate-stimulated release. Proceedings of the National Academy of Sciences of the United States of America 92:3948-3952. Schmid S, Hollmann M (2008) To Gate or not to Gate: Are the Delta Subunits in the Glutamate Receptor Family Functional Ion Channels? Molecular Neurobiology 37:126-141. Schorge S, Colquhoun D (2003) Studies of NMDA Receptor Function and Stoichiometry with Truncated and Tandem Subunits. J Neurosci 23:1151-1158. Schüler T, Mesic I, Madry C, Bartholomäus I, Laube B (2008) Formation of NR1/NR2 and NR1/NR3 Heterodimers Constitutes the Initial Step in NMethyl-D-aspartate Receptor Assembly. Journal of Biological Chemistry 283:37-46. Scott DB, Blanpied TA, Swanson GT, Zhang C, Ehlers MD (2001) An NMDA Receptor ER Retention Signal Regulated by Phosphorylation and Alternative Splicing. J Neurosci 21:3063-3072. Seeds NW, Williams BL, Bickford PC (1995) Tissue plasminogen activator induction in Purkinje neurons after cerebellar motor learning. Science 270:1992-1994. Seeds NW, Basham ME, Haffke SP (1999) Neuronal migration is retarded in mice lacking the tissue plasminogen activator gene. Proc Natl Acad Sci U S A 96:14118-14123. Seeds NW, Basham ME, Ferguson JE (2003) Absence of tissue plasminogen activator gene or activity impairs mouse cerebellar motor learning. J Neurosci 23:73687375. Sheinin A, Shavit S, Benveniste M (2001) Subunit specificity and mechanism of action of NMDA partial agonist D-cycloserine. Neuropharmacology 41:151158. Sheng M (2001) Molecular organization of the postsynaptic specialization. Proceedings of the National Academy of Sciences of the United States of America 98:7058-7061. Sheng M, Pak DTS (2000) Ligand-Gated Ion Channel Interactions with Cytoskeletal and Signaling Proteins. Annual Review of Physiology 62:755-778. Sheng M, Hoogenraad CC (2007) The postsynaptic architecture of excitatory synapses: a more quantitative view. Annu Rev Biochem 76:823-847. Shimizu C, Yoshida S, Shibata M, Kato K, Momota Y, Matsumoto K, Shiosaka T, Midorikawa R, Kamachi T, Kawabe A, Shiosaka S (1998) Characterization of recombinant and brain neuropsin, a plasticity-related serine protease. J Biol Chem 273:11189-11196. Shleper M, Kartvelishvily E, Wolosker H (2005) D-Serine Is the Dominant Endogenous Coagonist for NMDA Receptor Neurotoxicity in Organotypic Hippocampal Slices. J Neurosci 25:9413-9417. Simpkins KL, Guttmann RP, Dong Y, Chen Z, Sokol S, Neumar RW, Lynch DR (2003) Selective Activation Induced Cleavage of the NR2B Subunit by Calpain. J Neurosci 23:11322-11331. Skeberdis VA, Chevaleyre V, Lau CG, Goldberg JH, Pettit DL, Suadicani SO, Lin Y, Bennett MVL, Yuste R, Castillo PE, Zukin RS (2006) Protein kinase A regulates calcium permeability of NMDA receptors. Nat Neurosci 9:501-510. 145 Skrzypiec AE, Buczko W, Pawlak R (2008) Tissue plasminogen activator in the amygdala: a new role for an old protease. J Physiol Pharmacol 59 Suppl 8:135-146. Smothers CT, Woodward JJ (2007) Pharmacological characterization of glycineactivated currents in HEK 293 cells expressing N-methyl-D-aspartate NR1 and NR3 subunits. J Pharmacol Exp Ther 322:739-748. Smothers CT, Woodward JJ (2009) Expression of glycine-activated diheteromeric NR1/NR3 receptors in human embryonic kidney 293 cells Is NR1 splice variant-dependent. J Pharmacol Exp Ther 331:975-984. Snyder EM, Nong Y, Almeida CG, Paul S, Moran T, Choi EY, Nairn AC, Salter MW, Lombroso PJ, Gouras GK, Greengard P (2005) Regulation of NMDA receptor trafficking by amyloid-beta. Nat Neurosci 8:1051-1058. Sobolevsky AI, Yelshansky MV, Wollmuth LP (2004) The Outer Pore of the Glutamate Receptor Channel Has 2-Fold Rotational Symmetry. Neuron 41:367-378. Sobolevsky AI, Rosconi MP, Gouaux E (2009) X-ray structure, symmetry and mechanism of an AMPA-subtype glutamate receptor. Nature 462:745-756. Standley S, Roche KW, McCallum J, Sans N, Wenthold RJ (2000) PDZ Domain Suppression of an ER Retention Signal in NMDA Receptor NR1 Splice Variants. Neuron 28:887-898. Stefansson S, Muhammad S, Cheng XF, Battey FD, Strickland DK, Lawrence DA (1998) Plasminogen activator inhibitor-1 contains a cryptic high affinity binding site for the low density lipoprotein receptor-related protein. J Biol Chem 273:6358-6366. Strickland DK, Ranganathan S (2003) Diverse role of LDL receptor-related protein in the clearance of proteases and in signaling. J Thromb Haemost 1:1663-1670. Su EJ, Fredriksson L, Geyer M, Folestad E, Cale J, Andrae J, Gao Y, Pietras K, Mann K, Yepes M, Strickland DK, Betsholtz C, Eriksson U, Lawrence DA (2008) Activation of PDGF-CC by tissue plasminogen activator impairs blood-brain barrier integrity during ischemic stroke. Nat Med 14:731-737. Suchanek B, Seeburg PH, Sprengel R (1995) Gene Structure of the Murine N-Methyl D-Aspartate Receptor Subunit NR2C. Journal of Biological Chemistry 270:41-44. Sucher N, Akbarian S, Chi C, Leclerc C, Awobuluyi M, Deitcher D, Wu M, Yuan J, Jones E, Lipton S (1995) Developmental and regional expression pattern of a novel NMDA receptor- like subunit (NMDAR-L) in the rodent brain. J Neurosci 15:6509-6520. Sun Y, Olson R, Horning M, Armstrong N, Mayer M, Gouaux E (2002) Mechanism of glutamate receptor desensitization. Nature 417:245-253. Sutcliffe MJ, Wo ZG, Oswald RE (1996) Three-dimensional models of non-NMDA glutamate receptors. Biophysical Journal 70:1575-1589. Szklarczyk A, Ewaleifoh O, Beique JC, Wang Y, Knorr D, Haughey N, Malpica T, Mattson MP, Huganir R, Conant K (2008) MMP-7 cleaves the NR1 NMDA receptor subunit and modifies NMDA receptor function. FASEB J 22:37573767. Tabish M, Ticku MK (2004) Alternate splice variants of mouse NR2B gene. Neurochemistry International 44:339-343. Tang CM, Dichter M, Morad M (1990) Modulation of the N-methyl-D-aspartate channel by extracellular H+. Proceedings of the National Academy of Sciences of the United States of America 87:6445-6449. 146 Taverna FA, Wang LY, MacDonald JF, Hampson DR (1994) A transmembrane model for an ionotropic glutamate receptor predicted on the basis of the location of asparagine-linked oligosaccharides. Journal of Biological Chemistry 269:14159-14164. The National Institute of Neurological Disorders and Stroke rt-PA Stroke Study Group (1995) Tissue plasminogen activator for acute ischemic stroke. . N Engl J Med 333:1581-1587. Tong G, Takahashi H, Tu S, Shin Y, Talantova M, Zago W, Xia P, Nie Z, Goetz T, Zhang D, Lipton SA, Nakanishi N (2008) Modulation of NMDA Receptor Properties and Synaptic Transmission by the NR3A Subunit in Mouse Hippocampal and Cerebrocortical Neurons. J Neurophysiol 99:122-132. Tong W, Chen W, Ostrowski RP, Ma Q, Souvenir R, Zhang L, Zhang JH, Tang J (2010) Maternal hypoxia increases the activity of MMPs and decreases the expression of TIMPs in the brain of neonatal rats. Dev Neurobiol 70:182-194. Tovar KR, Westbrook GL (2002) Mobile NMDA receptors at hippocampal synapses. Neuron 34:255-264. Traynelis S, Hartley M, Heinemann S (1995) Control of proton sensitivity of the NMDA receptor by RNA splicing and polyamines. Science 268:873-876. Traynelis SF, Cull-Candy SG (1990) Proton inhibition of N-methyl-D-aspartate receptors in cerebellar neurons. Nature 345:347-350. Traynelis SF, Wollmuth LP, McBain CJ, Menniti FS, Vance KM, Ogden KK, Hansen KB, Yuan H, Myers SJ, Dingledine R (2010) Glutamate receptor ion channels: structure, regulation, and function. Pharmacol Rev 62:405-496. Triller A, Choquet D (2005) Surface trafficking of receptors between synaptic and extrasynaptic membranes: and yet they move! Trends Neurosci 28:133-139. Tsirka SE, Rogove AD, Bugge TH, Degen JL, Strickland S (1997) An Extracellular Proteolytic Cascade Promotes Neuronal Degeneration in the Mouse Hippocampus. J Neurosci 17:543-552. Turk B (2006) Targeting proteases: successes, failures and future prospects. Nat Rev Drug Discov 5:785-799. Ulbrich MH, Isacoff EY (2008) Rules of engagement for NMDA receptor subunits. Proceedings of the National Academy of Sciences 105:14163-14168. van Gent D, Sharp P, Morgan K, Kalsheker N (2003) Serpins: structure, function and molecular evolution. Int J Biochem Cell Biol 35:1536-1547. van Zundert B, Yoshii A, Constantine-Paton M (2004) Receptor compartmentalization and trafficking at glutamate synapses: a developmental proposal. Trends Neurosci 27:428-437. Vanaman TC, Bradshaw RA (1999) Proteases in cellular regulation minireview series. J Biol Chem 274:20047. VanDongen AM (2009) Biology of the NMDA receptor. Boca Raton: CRC Press. Vanhoutte P, Bading H (2003) Opposing roles of synaptic and extrasynaptic NMDA receptors in neuronal calcium signalling and BDNF gene regulation. Curr Opin Neurobiol 13:366-371. Vassalli JD, Sappino AP, Belin D (1991) The plasminogen activator/plasmin system. J Clin Invest 88:1067-1072. Vicini S, Wang JF, Li JH, Zhu WJ, Wang YH, Luo JH, Wolfe BB, Grayson DR (1998) Functional and Pharmacological Differences Between Recombinant NMethyl-D-Aspartate Receptors. J Neurophysiol 79:555-566. Villmann C, Becker CM (2007) On the hypes and falls in neuroprotection: targeting the NMDA receptor. Neuroscientist 13:594-615. 147 Vyklicky L, Jr., Vlachova V, Krusek J (1990) The effect of external pH changes on responses to excitatory amino acids in mouse hippocampal neurones. J Physiol 430:497-517. Wang YF, Tsirka SE, Strickland S, Stieg PE, Soriano SG, Lipton SA (1998) Tissue plasminogen activator (tPA) increase neuronal damage after focal cerebral ischemia in wild-type and tPA-deficient mice. Nat Med 4:228-231. Wang YT, Salter MW (1994) Regulation of NMDA receptors by tyrosine kinases and phosphatases. Nature 369:233-235. Wang YT, Yu XM, Salter MW (1996) Ca(2+)-independent reduction of N-methyl-Daspartate channel activity by protein tyrosine phosphatase. Proceedings of the National Academy of Sciences of the United States of America 93:1721-1725. Wee KS-L, Zhang Y, Khanna S, Low C-M (2008) Immunolocalization of NMDA receptor subunit NR3B in selected structures in the rat forebrain, cerebellum, and lumbar spinal cord. The Journal of Comparative Neurology 509:118-135. Wee XK, Ng KS, Leung HW, Cheong YP, Kong KH, Ng FM, Soh W, Lam Y, Low CM (2010) Mapping the high-affinity binding domain of 5-substituted benzimidazoles to the proximal N-terminus of the GluN2B subunit of the NMDA receptor. Br J Pharmacol 159:449-461. Wenzel A, Villa M, Mohler H (1996) Developmental and Regional Expression of NMDA Receptor Subtypes Containing the NR2D Subunit in Rat Brain. Journal of Neurochemistry 66:1240-1248. Westbrook GL, Mayer ML (1987) Micromolar concentrations of Zn2+ antagonize NMDA and GABA responses of hippocampal neurons. Nature 328:640-643. Williams K (1993) Ifenprodil discriminates subtypes of the N-methyl-D-aspartate receptor: selectivity and mechanisms at recombinant heteromeric receptors. Mol Pharmacol 44:851-859. Williams K (1994) Mechanisms influencing stimulatory effects of spermine at recombinant N-methyl-D-aspartate receptors. Molecular Pharmacology 46:161-168. Williams K (1996) Separating dual effects of zinc at recombinant N-methyl--aspartate receptors. Neuroscience Letters 215:9-12. Williams K, Kashiwagi K, Fukuchi J, Igarashi K (1995) An acidic amino acid in the N-methyl-D-aspartate receptor that is important for spermine stimulation. Mol Pharmacol 48:1087-1098. Williams K, Zappia AM, Pritchett DB, Shen YM, Molinoff PB (1994) Sensitivity of the N-methyl-D-aspartate receptor to polyamines is controlled by NR2 subunits. Mol Pharmacol 45:803-809. Willnow TE, Orth K, Herz J (1994) Molecular dissection of ligand binding sites on the low density lipoprotein receptor-related protein. J Biol Chem 269:1582715832. Wong E, Ng FM, Yu CY, Lim P, Lim LH, Traynelis SF, Low CM (2005) Expression and characterization of soluble amino-terminal domain of NR2B subunit of Nmethyl-D-aspartate receptor. Protein Sci 14:2275-2283. Wong H-K, Liu X-B, Matos MF, Chan SF, Pérez-Otaño I, Boysen M, Cui J, Nakanishi N, Trimmer JS, Jones EG, Lipton SA, Sucher NJ (2002) Temporal and regional expression of NMDA receptor subunit NR3A in the mammalian brain. The Journal of Comparative Neurology 450:303-317. Wu H-Y, Hsu F-C, Gleichman AJ, Baconguis I, Coulter DA, Lynch DR (2007) Fynmediated Phosphorylation of NR2B Tyr-1336 Controls Calpain-mediated 148 NR2B Cleavage in Neurons and Heterologous Systems. J Biol Chem 282:20075-20087. Wu HY, Yuen EY, Lu YF, Matsushita M, Matsui H, Yan Z, Tomizawa K (2005) Regulation of N-methyl-D-aspartate receptors by calpain in cortical neurons. J Biol Chem 280:21588-21593. Wu Z, Jiang G, Xiang P, Yang D, Wang N (2008) Purification and characterization of trypsin-like enzymes from North Pacific krill (Euphausia pacifica). Biotechnol Lett 30:67-72. Wyllie DJA, Béhé P, Colquhoun D (1998) Single-channel activations and concentration jumps: comparison of recombinant NR1a/NR2A and NR1a/NR2D NMDA receptors. The Journal of Physiology 510:1-18. Xu J, Kurup P, Zhang Y, Goebel-Goody SM, Wu PH, Hawasli AH, Baum ML, Bibb JA, Lombroso PJ (2009) Extrasynaptic NMDA receptors couple preferentially to excitotoxicity via calpain-mediated cleavage of STEP. J Neurosci 29:93309343. Yaka R, Thornton C, Vagts AJ, Phamluong K, Bonci A, Ron D (2002) NMDA receptor function is regulated by the inhibitory scaffolding protein, RACK1. Proc Natl Acad Sci U S A 99:5710-5715. Yamakura T, Shimoji K (1999) Subunit- and site-specific pharmacology of the NMDA receptor channel. Progress in Neurobiology 59:279-298. Yang CR, Svensson KA (2008) Allosteric modulation of NMDA receptor via elevation of brain glycine and d-serine: The therapeutic potentials for schizophrenia. Pharmacology & Therapeutics 120:317-332. Yepes M, Lawrence DA (2004) Neuroserpin: a selective inhibitor of tissue-type plasminogen activator in the central nervous system. Thromb Haemost 91:457-464. Yepes M, Roussel BD, Ali C, Vivien D (2009) Tissue-type plasminogen activator in the ischemic brain: more than a thrombolytic. Trends Neurosci 32:48-55. Yepes M, Sandkvist M, Moore EG, Bugge TH, Strickland DK, Lawrence DA (2003) Tissue-type plasminogen activator induces opening of the blood-brain barrier via the LDL receptor-related protein. J Clin Invest 112:1533-1540. Yong VW (2005) Metalloproteinases: mediators of pathology and regeneration in the CNS. Nat Rev Neurosci 6:931-944. Yu X-M, Askalan R, Keil GJ, II, Salter MW (1997) NMDA Channel Regulation by Channel-Associated Protein Tyrosine Kinase Src. Science 275:674-678. Yuan H, Erreger K, Dravid SM, Traynelis SF (2005) Conserved Structural and Functional Control of N-Methyl-d-aspartate Receptor Gating by Transmembrane Domain M3. Journal of Biological Chemistry 280:2970829716. Yuan H, Hansen KB, Vance KM, Ogden KK, Traynelis SF (2009a) Control of NMDA Receptor Function by the NR2 Subunit Amino-Terminal Domain. J Neurosci 29:12045-12058. Yuan H, Vance KM, Junge CE, Geballe MT, Snyder JP, Hepler JR, Yepes M, Low CM, Traynelis SF (2009b) The serine protease plasmin cleaves the aminoterminal domain of the NR2A subunit to relieve zinc inhibition of the Nmethyl-D-aspartate receptors. J Biol Chem 284:12862-12873. Yuen EY, Gu Z, Yan Z (2007) Calpain regulation of AMPA receptor channels in cortical pyramidal neurons. J Physiol 580:241-254. 149 Yuen EY, Ren Y, Yan Z (2008) Postsynaptic density-95 (PSD-95) and calcineurin control the sensitivity of N-methyl-D-aspartate receptors to calpain cleavage in cortical neurons. Mol Pharmacol 74:360-370. Yuzaki M (2003) The [delta]2 glutamate receptor: 10 years later. Neuroscience Research 46:11-22. Zhang L, Zheng X, Paupard MC, Wang AP, Santchi L, Friedman LK, Zukin RS, Bennett MV (1994) Spermine potentiation of recombinant N-methyl-Daspartate receptors is affected by subunit composition. Proc Natl Acad Sci U S A 91:10883-10887. Zhang S, Edelmann L, Liu J, Crandall JE, Morabito MA (2008) Cdk5 Regulates the Phosphorylation of Tyrosine 1472 NR2B and the Surface Expression of NMDA Receptors. J Neurosci 28:415-424. Zhuo M (2009) Plasticity of NMDA receptor NR2B subunit in memory and chronic pain. Mol Brain 2:4. Zhuo M, Holtzman DM, Li Y, Osaka H, DeMaro J, Jacquin M, Bu G (2000) Role of tissue plasminogen activator receptor LRP in hippocampal long-term potentiation. J Neurosci 20:542-549. Zukin RS, Bennett MVL (1995) Alternatively spliced isoforms of the NMDARI receptor subunit. Trends in Neurosciences 18:306-313. 150 [...]... heteromeric structure of the NMDA receptor is elucidated 1.4 NMDA Receptor Channel Properties and Pharmacology Unlike the AMPA and kainate receptors, binding of glutamate alone will not activate the NMDA receptors Activation of NMDA receptors is unique, as it requires the simultaneous binding of the agonist glutamate to the LBD of NR2 subunits and the co-agonist glycine to the LBD of NR1 subunits, in... tetrameric NMDA receptor structure and the topology of a single subunit (Left) The tetrameric NMDA receptor, a membrane receptor, is an assembly of the obligatory NR1 subunit and NR2 and/ or NR3 subunits The activation of NMDA receptors requires the binding of glutamate to NR2 subunits and glycine to NR1 subunit, together with the removal of Mg2+ upon membrane depolarization Activated NMDA receptors. .. used drugs amantadine and memantine (Yamakura and Shimoji, 1999; Kew and Kemp, 2005; Paoletti and Neyton, 2007) Although most of the channel blockers, such as PCP and ketamine, do not show subtype selectivity, some of them, such as Mg2+ and MK801, have higher affinity towards NR2A and NR2B containing NMDA receptors (Yamakura and Shimoji, 1999; Paoletti and Neyton, 2007) 1.5 NMDA Receptors at the Glutamatergic... proteins and contains various posttranslational modification sites 9 Studies of the NMDA receptor function and pharmacology have led to the discovery that a variety of subunit-specific ligands can target the ATD of NMDA receptors and regulate the receptor function allosterically (Paoletti and Neyton, 2007; Mony et al., 2009a) Antagonistic ligands, such as Zn2+ and ifenprodil, have been found to inhibit NMDA. .. 1999) 1.3 NMDA Receptors in the Brain: Localization and Architecture Named after the original agonists used to activate them selectively, NMDA receptors are a class of ligand-gated ion channels that are expressed in many parts of the brain in both neonatal and adult brains Activation of NMDA receptors allow influx of calcium ions (Ca2+) into neurons which will trigger various downstream events and signalling...List of figures Figure 1.1 Classification of glutamate receptors 2 Figure 1.2 The different isoforms of the NR1 subunit 5 Figure 1.3 The tetrameric NMDA receptor structure and the topology of a single subunit 9 Figure 1.4 The glutamatergic synapse 20 Figure 1.5 Phosphorylation sites residing in the CTD of NMDA receptor subunits 28 Figure 1.6 Differential surface regulation of NR2B-containing NMDA receptors. .. C1 and C2 cassettes, NR1-2 lacks the C1 cassette, NR1-3 lacks the C2 cassette and NR1-4 lacks both the C1 and C2 cassette The letter ‘a’ and ‘b’ indicate the absence and presence of exon 5 respectively (Hollmann et al., 1993) 4 The expression of different splice variants of the NR1 subunit are temporally and spatially regulated (Laurie and Seeburg, 1994a) The expression of most of the NR1 splice isoforms... nature, the NMDA receptors play essential roles in neuronal development, synaptic transmission and synaptic plasticity (Ulbrich and Isacoff, 2008) However, over-activation of NMDA receptors can lead to excessive Ca2+ influx and neuronal death due to excitotoxicity (Dirnagl et al., 1999) In addition to excitotoxicity, NMDA receptors have been implicated in many pathophysiological conditions and neurological... 1.4.2.1 Agonists Activation of NMDA receptors requires binding of both agonist L-glutamate and co-agonist glycine (Johnson and Ascher, 1987; Kleckner and Dingledine, 1988) In addition, L-aspartate has also been found to activate NMDA receptors, albeit at a lower affinity (~5 fold) compared to L-glutamate (Patneau and Mayer, 1990) The 15 binding of glutamate to NR2 subunits and glycine to NR1 subunits... protein SEM Standard error of the mean TEVC Two-electrode voltage-clamp tPA Tissue- type plasminogen activator uPA Urokinase-type plasminogen activator UTR Untranslated region xiv CHAPTER 1 Introduction CHAPTER 1 Introduction 1 1.1 Glutamate Receptors in the Mammalian Central Nervous System Neural transmission is a critical component of the many processes regulating the normal functioning of the mammalian . Title Page REGULATION OF NMDA RECEPTORS BY SERINE PROTEASES TISSUE PLASMINOGEN ACTIVATOR (tPA) AND PLASMINOGEN/ PLASMIN NG KAY SIONG B. Appl. Sci 30 1.6.3.3 Thrombin 30 1.6.3.4 Tissue- type plasminogen activator (tPA) 32 1.7 Tissue- type Plasminogen Activator (tPA) 32 1.7.1 tPA and stroke 32 1.7.2 tPA /plasminogen system in the brain. neurotoxicity through the NMDA receptors 37 1.8.3.2 Plasmin cleavage of NMDA receptors 38 1.8.3.3 LRP and the NMDA receptor 39 1.8.3.4 tPA and NR2B-containing NMDA receptors 40 1.9 Thesis