Top

Molecular Pain

Published in:

Open Access 01-12-2011 | Research

Lentiviral gene transfer into the dorsal root ganglion of adult rats

Authors: Hongwei Yu, Greg Fischer, Guangfu Jia, Jakob Reiser, Frank Park, Quinn H Hogan

Published in: Molecular Pain | Issue 1/2011

Abstract

Background

Lentivector-mediated gene delivery into the dorsal root ganglion (DRG) is a promising method for exploring pain pathophysiology and for genetic treatment of chronic neuropathic pain. In this study, a series of modified lentivector particles with different cellular promoters, envelope glycoproteins, and viral accessory proteins were generated to evaluate the requirements for efficient transduction into neuronal cells in vitro and adult rat DRG in vivo.

Results

In vitro, lentivectors expressing enhanced green fluorescent protein (EGFP) under control of the human elongation factor 1α (EF1α) promoter and pseudotyped with the conventional vesicular stomatitis virus G protein (VSV-G) envelope exhibited the best performance in the transfer of EGFP into an immortalized DRG sensory neuron cell line at low multiplicities of infection (MOIs), and into primary cultured DRG neurons at higher MOIs. In vivo, injection of either first or second-generation EF1α-EGFP lentivectors directly into adult rat DRGs led to transduction rates of 19 ± 9% and 20 ± 8% EGFP-positive DRG neurons, respectively, detected at 4 weeks post injection. Transduced cells included a full range of neuronal phenotypes, including myelinated neurons as well as both non-peptidergic and peptidergic nociceptive unmyelinated neurons.

Conclusion

VSV-G pseudotyped lentivectors containing the human elongation factor 1α (EF1α)-EGFP expression cassette demonstrated relatively efficient transduction to sensory neurons following direct injection into the DRG. These results clearly show the potential of lentivectors as a viable system for delivering target genes into DRGs to explore basic mechanisms of neuropathic pain, with the potential for future clinical use in treating chronic pain.

Available only for authorised users

Backonja M, Woolf CJ: Future directions in neuropathic pain therapy: closing the translational loop. Oncologist 2010,15(Suppl 2):24–29.PubMedCrossRef

Devor M, Seltzer Z: Pathology of damaged nerves in relation to chronic pain. In Textbook of Pain. 4th edition. Edited by: Wall PD, Melzack R. Edinburgh: Churchill Livingstone; 1999:129–164.

Hermens WT, Verhaagen J: Viral vectors, tools for gene transfer in the nervous system. Prog Neurobiol 1998,55(4):399–432. 10.1016/S0301-0082(98)00007-0PubMedCrossRef

Beutler AS, Reinhardt M: AAV for pain: steps towards clinical translation. Gene therapy 2009,16(4):461–469. 10.1038/gt.2009.23PubMedCrossRef

Glorioso JC, Fink DJ: Herpes vector-mediated gene transfer in the treatment of chronic pain. Mol Ther 2009,17(1):13–18. 10.1038/mt.2008.213PubMedCentralPubMedCrossRef

Storek B, Reinhardt M, Wang C, Janssen WG, Harder NM, Banck MS, Morrison JH, Beutler AS: Sensory neuron targeting by self-complementary AAV8 via lumbar puncture for chronic pain. Proceedings of the National Academy of Sciences of the United States of America 2008,105(3):1055–1060. 10.1073/pnas.0708003105PubMedCentralPubMedCrossRef

Huang X, Yang Y: Innate Immune Recognition of Viruses and Viral Vectors. Human gene therapy 2009.

Naldini L, Blomer U, Gallay P, Ory D, Mulligan R, Gage FH, Verma IM, Trono D: In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 1996,272(5259):263–267. 10.1126/science.272.5259.263PubMedCrossRef

Georgievska B, Kirik D, Bjorklund A: Overexpression of glial cell line-derived neurotrophic factor using a lentiviral vector induces time- and dose-dependent downregulation of tyrosine hydroxylase in the intact nigrostriatal dopamine system. J Neurosci 2004,24(29):6437–6445. 10.1523/JNEUROSCI.1122-04.2004PubMedCrossRef

10.

Pezet S, Krzyzanowska A, Wong LF, Grist J, Mazarakis ND, Georgievska B, McMahon SB: Reversal of neurochemical changes and pain-related behavior in a model of neuropathic pain using modified lentiviral vectors expressing GDNF. Mol Ther 2006,13(6):1101–1109. 10.1016/j.ymthe.2005.11.026PubMedCrossRef

11.

Dittgen T, Nimmerjahn A, Komai S, Licznerski P, Waters J, Margrie TW, Helmchen F, Denk W, Brecht M, Osten P: Lentivirus-based genetic manipulations of cortical neurons and their optical and electrophysiological monitoring in vivo. Proceedings of the National Academy of Sciences of the United States of America 2004,101(52):18206–18211. 10.1073/pnas.0407976101PubMedCentralPubMedCrossRef

12.

Ehrengruber MU, Hennou S, Bueler H, Naim HY, Deglon N, Lundstrom K: Gene transfer into neurons from hippocampal slices: comparison of recombinant Semliki Forest Virus, adenovirus, adeno-associated virus, lentivirus, and measles virus. Molecular and cellular neurosciences 2001,17(5):855–871. 10.1006/mcne.2001.0982PubMedCrossRef

13.

Torashima T, Okoyama S, Nishizaki T, Hirai H: In vivo transduction of murine cerebellar Purkinje cells by HIV-derived lentiviral vectors. Brain research 2006,1082(1):11–22. 10.1016/j.brainres.2006.01.104PubMedCrossRef

14.

Eggers R, Hendriks WT, Tannemaat MR, van Heerikhuize JJ, Pool CW, Carlstedt TP, Zaldumbide A, Hoeben RC, Boer GJ, Verhaagen J: Neuroregenerative effects of lentiviral vector-mediated GDNF expression in reimplanted ventral roots. Molecular and cellular neurosciences 2008,39(1):105–117. 10.1016/j.mcn.2008.05.018PubMedCrossRef

15.

Fleming J, Ginn SL, Weinberger RP, Trahair TN, Smythe JA, Alexander IE: Adeno-associated virus and lentivirus vectors mediate efficient and sustained transduction of cultured mouse and human dorsal root ganglia sensory neurons. Human gene therapy 2001,12(1):77–86. 10.1089/104303401450997PubMedCrossRef

16.

Mikami M, Yang J: Short hairpin RNA-mediated selective knockdown of NaV1.8 tetrodotoxin-resistant voltage-gated sodium channel in dorsal root ganglion neurons. Anesthesiology 2005,103(4):828–836. 10.1097/00000542-200510000-00022PubMedCrossRef

17.

Park F, Ohashi K, Chiu W, Naldini L, Kay MA: Efficient lentiviral transduction of liver requires cell cycling in vivo. Nat Genet 2000,24(1):49–52. 10.1038/71673PubMedCrossRef

18.

Pfirrmann CW, Oberholzer PA, Zanetti M, Boos N, Trudell DJ, Resnick D, Hodler J: Selective nerve root blocks for the treatment of sciatica: evaluation of injection site and effectiveness--a study with patients and cadavers. Radiology 2001,221(3):704–711. 10.1148/radiol.2213001635PubMedCrossRef

19.

Fischer G, Kostic S, Nakai H, Park F, Sapunar D, Yu H, Hogan Q: Direct injection into the dorsal root ganglion: Technical, behavioral, and histological observations. Journal of neuroscience methods

20.

Mazarakis ND, Azzouz M, Rohll JB, Ellard FM, Wilkes FJ, Olsen AL, Carter EE, Barber RD, Baban DF, Kingsman SM, et al.: Rabies virus glycoprotein pseudotyping of lentiviral vectors enables retrograde axonal transport and access to the nervous system after peripheral delivery. Hum Mol Genet 2001,10(19):2109–2121. 10.1093/hmg/10.19.2109PubMedCrossRef

21.

Chen W, Mi R, Haughey N, Oz M, Hoke A: Immortalization and characterization of a nociceptive dorsal root ganglion sensory neuronal line. J Peripher Nerv Syst 2007,12(2):121–130. 10.1111/j.1529-8027.2007.00131.xPubMedCentralPubMedCrossRef

22.

Vetter I, Lewis RJ: Characterization of endogenous calcium responses in neuronal cell lines. Biochem Pharmacol 79(6):908–920.

23.

Kostic C, Chiodini F, Salmon P, Wiznerowicz M, Deglon N, Hornfeld D, Trono D, Aebischer P, Schorderet DF, Munier FL, et al.: Activity analysis of housekeeping promoters using self-inactivating lentiviral vector delivery into the mouse retina. Gene therapy 2003,10(9):818–821. 10.1038/sj.gt.3301948PubMedCrossRef

24.

Li M, Husic N, Lin Y, Christensen H, Malik I, McIver S, Daniels CM, Harris DA, Kotzbauer PT, Goldberg MP, et al.: Optimal promoter usage for lentiviral vector-mediated transduction of cultured central nervous system cells. Journal of neuroscience methods 2010,189(1):56–64. 10.1016/j.jneumeth.2010.03.019PubMedCentralPubMedCrossRef

25.

Papadakis ED, Nicklin SA, Baker AH, White SJ: Promoters and control elements: designing expression cassettes for gene therapy. Current gene therapy 2004,4(1):89–113. 10.2174/1566523044578077PubMedCrossRef

26.

Haastert K, Mauritz C, Chaturvedi S, Grothe C: Human and rat adult Schwann cell cultures: fast and efficient enrichment and highly effective non-viral transfection protocol. Nat Protoc 2007,2(1):99–104. 10.1038/nprot.2006.486PubMedCrossRef

27.

Hogan Q, Sapunar D, Modric-Jednacak K, McCallum JB: Detection of neuropathic pain in a rat model of peripheral nerve injury. Anesthesiology 2004,101(2):476–487. 10.1097/00000542-200408000-00030PubMedCrossRef

28.

Wu HE, Gemes G, Zoga V, Kawano T, Hogan QH: Learned avoidance from noxious mechanical simulation but not threshold semmes weinstein filament stimulation after nerve injury in rats. J Pain 2010,11(3):280–286. 10.1016/j.jpain.2009.07.011PubMedCentralPubMedCrossRef

29.

Finnerup NB, Otto M, McQuay HJ, Jensen TS, Sindrup SH: Algorithm for neuropathic pain treatment: an evidence based proposal. Pain 2005,118(3):289–305. 10.1016/j.pain.2005.08.013PubMedCrossRef

30.

Jimenez-Andrade JM, Herrera MB, Ghilardi JR, Vardanyan M, Melemedjian OK, Mantyh PW: Vascularization of the dorsal root ganglia and peripheral nerve of the mouse: implications for chemical-induced peripheral sensory neuropathies. Mol Pain 2008, 4: 10. 10.1186/1744-8069-4-10PubMedCentralPubMedCrossRef

31.

Costigan M, Scholz J, Woolf CJ: Neuropathic pain: a maladaptive response of the nervous system to damage. Annu Rev Neurosci 2009, 32: 1–32. 10.1146/annurev.neuro.051508.135531PubMedCentralPubMedCrossRef

32.

Iwamoto N, Watanabe A, Yamamoto M, Miyake N, Kurai T, Teramoto A, Shimada T: Global diffuse distribution in the brain and efficient gene delivery to the dorsal root ganglia by intrathecal injection of adeno-associated viral vector serotype 1. J Gene Med 2009,11(6):498–505. 10.1002/jgm.1325PubMedCrossRef

33.

Towne C, Pertin M, Beggah AT, Aebischer P, Decosterd I: Recombinant adeno-associated virus serotype 6 (rAAV2/6)-mediated gene transfer to nociceptive neurons through different routes of delivery. Mol Pain 2009, 5: 52. 10.1186/1744-8069-5-52PubMedCentralPubMedCrossRef

34.

Xu Y, Gu Y, Wu P, Li GW, Huang LY: Efficiencies of transgene expression in nociceptive neurons through different routes of delivery of adeno-associated viral vectors. Human gene therapy 2003,14(9):897–906. 10.1089/104303403765701187PubMedCrossRef

35.

Mason MR, Ehlert EM, Eggers R, Pool CW, Hermening S, Huseinovic A, Timmermans E, Blits B, Verhaagen J: Comparison of AAV serotypes for gene delivery to dorsal root ganglion neurons. Molecular therapy: the journal of the American Society of Gene Therapy 2010,18(4):715–724.CrossRef

36.

Glatzel M, Flechsig E, Navarro B, Klein MA, Paterna JC, Bueler H, Aguzzi A: Adenoviral and adeno-associated viral transfer of genes to the peripheral nervous system. Proceedings of the National Academy of Sciences of the United States of America 2000,97(1):442–447. 10.1073/pnas.97.1.442PubMedCentralPubMedCrossRef

37.

Nayak S, Herzog RW: Progress and prospects: immune responses to viral vectors. Gene Ther 2010,17(3):295–304. 10.1038/gt.2009.148PubMedCentralPubMedCrossRef

38.

Ehrhardt A, Kay MA: A new adenoviral helper-dependent vector results in long-term therapeutic levels of human coagulation factor IX at low doses in vivo. Blood 2002,99(11):3923–3930. 10.1182/blood.V99.11.3923PubMedCrossRef

39.

Ehrhardt A, Xu H, Dillow AM, Bellinger DA, Nichols TC, Kay MA: A gene-deleted adenoviral vector results in phenotypic correction of canine hemophilia B without liver toxicity or thrombocytopenia. Blood 2003,102(7):2403–2411. 10.1182/blood-2003-01-0314PubMedCrossRef

40.

Kochanek S, Schiedner G, Volpers C: High-capacity 'gutless' adenoviral vectors. Curr Opin Mol Ther 2001,3(5):454–463.PubMed

41.

Terashima T, Oka K, Kritz AB, Kojima H, Baker AH, Chan L: DRG-targeted helper-dependent adenoviruses mediate selective gene delivery for therapeutic rescue of sensory neuronopathies in mice. The Journal of clinical investigation 2009,119(7):2100–2112.PubMedCentralPubMed

42.

Glorioso JC, Fink DJ: Gene therapy for pain: introduction to the special issue. Gene Ther 2009,16(4):453–454. 10.1038/gt.2009.18PubMedCrossRef

43.

Monville C, Torres E, Thomas E, Scarpini CG, Muhith J, Lewis J, Finn J, Smith C, Cai S, Efstathiou S, et al.: HSV vector-delivery of GDNF in a rat model of PD: partial efficacy obscured by vector toxicity. Brain research 2004,1024(12):1–15.PubMedCrossRef

44.

Mason MR, Ehlert EM, Eggers R, Pool CW, Hermening S, Huseinovic A, Timmermans E, Blits B, Verhaagen J: Comparison of AAV Serotypes for Gene Delivery to Dorsal Root Ganglion Neurons. Mol Ther 2010.

45.

Park F, Ohashi K, Kay MA: Therapeutic levels of human factor VIII and IX using HIV-1-based lentiviral vectors in mouse liver. Blood 2000,96(3):1173–1176.PubMed

46.

Escors D, Breckpot K: Lentiviral Vectors in Gene Therapy: Their Current Status and Future Potential. Arch Immunol Ther Exp (Warsz) 2010.

47.

Meunier A, Pohl M: Lentiviral vectors for gene transfer into the spinal cord glial cells. Gene therapy 2009,16(4):476–482. 10.1038/gt.2009.22PubMedCrossRef

48.

Kumar M, Keller B, Makalou N, Sutton RE: Systematic determination of the packaging limit of lentiviral vectors. Hum Gene Ther 2001,12(15):1893–1905. 10.1089/104303401753153947PubMedCrossRef

49.

Oh T, Peister A, Ohashi K, Park F: Transplantation of murine bone marrow stromal cells under the kidney capsule to secrete coagulation factor VIII. Cell Transplant 2006,15(7):637–645. 10.3727/000000006783981620PubMedCrossRef

50.

Dettenhofer M, Yu XF: Highly purified human immunodeficiency virus type 1 reveals a virtual absence of Vif in virions. J Virol 1999,73(2):1460–1467.PubMedCentralPubMed

51.

Hacein-Bey-Abina S, Garrigue A, Wang GP, Soulier J, Lim A, Morillon E, Clappier E, Caccavelli L, Delabesse E, Beldjord K, et al.: Insertional oncogenesis in 4 patients after retrovirus-mediated gene therapy of SCID-X1. J Clin Invest 2008,118(9):3132–3142. 10.1172/JCI35700PubMedCentralPubMedCrossRef

52.

Hacein-Bey-Abina S, von Kalle C, Schmidt M, Le Deist F, Wulffraat N, McIntyre E, Radford I, Villeval JL, Fraser CC, Cavazzana-Calvo M, et al.: A serious adverse event after successful gene therapy for X-linked severe combined immunodeficiency. N Engl J Med 2003,348(3):255–256. 10.1056/NEJM200301163480314PubMedCrossRef

53.

Hacein-Bey-Abina S, Von Kalle C, Schmidt M, McCormack MP, Wulffraat N, Leboulch P, Lim A, Osborne CS, Pawliuk R, Morillon E, et al.: LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science 2003,302(5644):415–419. 10.1126/science.1088547PubMedCrossRef

54.

Beard BC, Dickerson D, Beebe K, Gooch C, Fletcher J, Okbinoglu T, Miller DG, Jacobs MA, Kaul R, Kiem HP, et al.: Comparison of HIV-derived lentiviral and MLV-based gammaretroviral vector integration sites in primate repopulating cells. Mol Ther 2007,15(7):1356–1365. 10.1038/sj.mt.6300159PubMedCrossRef

55.

Nakai H, Montini E, Fuess S, Storm TA, Grompe M, Kay MA: AAV serotype 2 vectors preferentially integrate into active genes in mice. Nat Genet 2003,34(3):297–302. 10.1038/ng1179PubMedCrossRef

56.

Donsante A, Miller DG, Li Y, Vogler C, Brunt EM, Russell DW, Sands MS: AAV vector integration sites in mouse hepatocellular carcinoma. Science 2007,317(5837):477. 10.1126/science.1142658PubMedCrossRef

57.

Scherr M, Battmer K, Eder M, Schule S, Hohenberg H, Ganser A, Grez M, Blomer U: Efficient gene transfer into the CNS by lentiviral vectors purified by anion exchange chromatography. Gene therapy 2002,9(24):1708–1714. 10.1038/sj.gt.3301848PubMedCrossRef

58.

Hogan Q: Injection for diagnosis and therapy of back disease, Principles and Techniques of Spine Surgery. Edited by: An H. Baltimore (MD): Williams and Willkins; 1998:707–729.

59.

Pfeifer A, Ikawa M, Dayn Y, Verma IM: Transgenesis by lentiviral vectors: lack of gene silencing in mammalian embryonic stem cells and preimplantation embryos. Proc Natl Acad Sci USA 2002,99(4):2140–2145. 10.1073/pnas.251682798PubMedCentralPubMedCrossRef

60.

Park F, Sweeney WE, Jia G, Roman RJ, Avner ED: 20-HETE mediates proliferation of renal epithelial cells in polycystic kidney disease. J Am Soc Nephrol 2008,19(10):1929–1939. 10.1681/ASN.2007070771PubMedCentralPubMedCrossRef

61.

Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, Naldini L: A third-generation lentivirus vector with a conditional packaging system. J Virol 1998,72(11):8463–8471.PubMedCentralPubMed

62.

Beyer WR, Westphal M, Ostertag W, von Laer D: Oncoretrovirus and lentivirus vectors pseudotyped with lymphocytic choriomeningitis virus glycoprotein: generation, concentration, and broad host range. J Virol 2002,76(3):1488–1495. 10.1128/JVI.76.3.1488-1495.2002PubMedCentralPubMedCrossRef

63.

Federici T, Kutner R, Zhang XY, Kuroda H, Tordo N, Boulis NM, Reiser J: Comparative analysis of HIV-1-based lentiviral vectors bearing lyssavirus glycoproteins for neuronal gene transfer. Genetic vaccines and therapy 2009, 7: 1. 10.1186/1479-0556-7-1PubMedCentralPubMedCrossRef

64.

Conzelmann KK, Cox JH, Schneider LG, Thiel HJ: Molecular cloning and complete nucleotide sequence of the attenuated rabies virus SAD B19. Virology 1990,175(2):485–499. 10.1016/0042-6822(90)90433-RPubMedCrossRef

65.

Park F, Kay MA: Modified HIV-1 based lentiviral vectors have an effect on viral transduction efficiency and gene expression in vitro and in vivo. Mol Ther 2001,4(3):164–173. 10.1006/mthe.2001.0450PubMedCrossRef

66.

Jorgensen NP, Nossen JO, Borch KW, Kristiansen AB, Kristoffersen DT, Lundby B, Theodorsen L: Safety and tolerability of iodixanol in healthy volunteers with reference to two monomeric X-ray contrast media. Eur J Radiol 1992,15(3):252–257. 10.1016/0720-048X(92)90118-SPubMedCrossRef

67.

Malin SA, Davis BM, Molliver DC: Production of dissociated sensory neuron cultures and considerations for their use in studying neuronal function and plasticity. Nat Protoc 2007,2(1):152–160. 10.1038/nprot.2006.461PubMedCrossRef

68.

Baekelandt V, Claeys A, Eggermont K, Lauwers E, De Strooper B, Nuttin B, Debyser Z: Characterization of lentiviral vector-mediated gene transfer in adult mouse brain. Human gene therapy 2002,13(7):841–853. 10.1089/10430340252899019PubMedCrossRef

69.

Carbonaro DA, Jin X, Petersen D, Wang X, Dorey F, Kil KS, Aldrich M, Blackburn MR, Kellems RE, Kohn DB: In vivo transduction by intravenous injection of a lentiviral vector expressing human ADA into neonatal ADA gene knockout mice: a novel form of enzyme replacement therapy for ADA deficiency. Mol Ther 2006,13(6):1110–1120. 10.1016/j.ymthe.2006.02.013PubMedCrossRef

70.

Lobbestael E, Reumers V, Ibrahimi A, Paesen K, Thiry I, Gijsbers R, Van den Haute C, Debyser Z, Baekelandt V, Taymans JM: Immunohistochemical detection of transgene expression in the brain using small epitope tags. BMC biotechnology 10: 16.

71.

Yu H, Wessels A, Chen J, Phelps AL, Oatis J, Tint GS, Patel SB: Late gestational lung hypoplasia in a mouse model of the Smith-Lemli-Opitz syndrome. BMC Dev Biol 2004, 4: 1. 10.1186/1471-213X-4-1PubMedCentralPubMedCrossRef

72.

Yu H, Wessels A, Tint GS, Patel SB: Partial rescue of neonatal lethality of Dhcr7 null mice by a nestin promoter-driven DHCR7 transgene expression. Brain Res Dev Brain Res 2005,156(1):46–60.PubMedCrossRef

73.

Jin SX, Zhuang ZY, Woolf CJ, Ji RR: p38 mitogen-activated protein kinase is activated after a spinal nerve ligation in spinal cord microglia and dorsal root ganglion neurons and contributes to the generation of neuropathic pain. J Neurosci 2003,23(10):4017–4022.PubMed

74.

Molliver DC, Wright DE, Leitner ML, Parsadanian AS, Doster K, Wen D, Yan Q, Snider WD: IB4-binding DRG neurons switch from NGF to GDNF dependence in early postnatal life. Neuron 1997,19(4):849–861. 10.1016/S0896-6273(00)80966-6PubMedCrossRef

75.

White FA, Sun J, Waters SM, Ma C, Ren D, Ripsch M, Steflik J, Cortright DN, Lamotte RH, Miller RJ: Excitatory monocyte chemoattractant protein-1 signaling is up-regulated in sensory neurons after chronic compression of the dorsal root ganglion. Proceedings of the National Academy of Sciences of the United States of America 2005,102(39):14092–14097. 10.1073/pnas.0503496102PubMedCentralPubMedCrossRef

Title: Lentiviral gene transfer into the dorsal root ganglion of adult rats
Authors: Hongwei Yu
Greg Fischer
Guangfu Jia
Jakob Reiser
Frank Park
Quinn H Hogan
Publication date: 01-12-2011
Publisher: BioMed Central
Published in: Molecular Pain / Issue 1/2011
Electronic ISSN: 1744-8069
DOI: https://doi.org/10.1186/1744-8069-7-63

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Lentiviral gene transfer into the dorsal root ganglion of adult rats

Abstract

Background

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2011

Rapid S-nitrosylation of actin by NO-generating donors and in inflammatory pain model mice

Involvement of the Cav3.2 T-type calcium channel in thalamic neuron discharge patterns

Enhanced inhibitory synaptic transmission in the spinal dorsal horn mediates antinociceptive effects of TC-2559

Resveratrol inhibits Cdk5 activity through regulation of p35 expression

Specific involvement of atypical PKCζ/PKMζ in spinal persistent nociceptive processing following peripheral inflammation in rat

The analgesic effect of electroacupuncture on acute thermal pain perception-a central neural correlate study with fMRI