title: Slice culture modeling of central nervous system (CNS) viral infection.
abstract: The complexity of the central nervous system (CNS) is not recapitulated in cell culture models. Thin slicing and subsequent culture of CNS tissue has become a valued means to study neuronal and glial biology within the context of the physiologically relevant tissue milieu. Modern membrane-interface slice culturing methodology allows straightforward access to both CNS tissue and feeding medium, enabling experimental manipulations and analyses that would otherwise be impossible in vivo. CNS slices can be successfully maintained in culture for up to several weeks for investigation of evolving pathology and long-term intervention in models of chronic neurologic disease.Herein, membrane-interface slice culture models for studying viral encephalitis and myelitis are detailed, with emphasis on the use of these models for investigation of pathogenesis and evaluation of novel treatment strategies. We describe techniques to (1) generate brain and spinal cord slices from rodent donors, (2) virally infect slices, (3) assess virally induced injury/apoptosis, (4) characterize "CNS-specific" cytokine production, and (5) treat slices with cytokines/pharmaceuticals. Although our focus is on CNS viral infection, we anticipate that the described methods can be adapted to address a wide range of investigations within the fields of neuropathology, neuroimmunology, and neuropharmacology.
authors: Department of Neurology, Denver Veterans Affairs Medical Center, Aurora, CO, USA.
authors: Kalen R
authors: KR
authors: Dionne
authors: Kenneth L
authors: KL
authors: Tyler
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism / Q000473:pathology*; D002021:Buffers; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D016616:Cryoultramicrotomy; D016207:Cytokines / Q000096:biosynthesis / Q000494:pharmacology; D004660:Encephalitis / Q000378:metabolism / Q000473:pathology*; D005455:Fluorescent Antibody Technique; D006836:Hydro-Lyases / Q000378:metabolism; D051379:Mice; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D011419:Propidium / Q000378:metabolism; D013116:Spinal Cord / Q000378:metabolism / Q000473:pathology*; D046509:Tissue Culture Techniques
chemical_list: D002021:Buffers; D016207:Cytokines; D011419:Propidium; D020169:Caspases; D006836:Hydro-Lyases; C045581:lactate dehydratase
references: Neuroscience. 1999;94(3):755-65
references: 10579566
references: J Neurotrauma. 2004 Aug;21(8):1103-12
references: 15319009
references: J Gen Virol. 2006 Oct;87(Pt 10):2827-2837
references: 16963740
references: Behav Brain Res. 2008 Sep 1;192(1):106-13
references: 18359102
references: J Neurosci Methods. 1981 Dec;4(4):329-42
references: 7033675
references: J Virol. 2002 Jul;76(14):7247-54
references: 12072524
references: Neuroscience. 2000;96(1):131-9
references: 10683418
references: Curr Neuropharmacol. 2007 Mar;5(1):19-33
references: 18615151
references: Brain Pathol. 1993 Oct;3(4):413-9
references: 8293197
references: Neurochem Res. 2005 Dec;30(12):1521-8
references: 16362771
references: Drug Discov Today. 2005 Jul 15;10(14):993-1000
references: 16023058
references: Brain Res. 1976 May 28;108(2):423-30
references: 6116
references: Brain Res Brain Res Protoc. 2000 Apr;5(2):153-8
references: 10775835
references: Am J Anat. 1951 Nov;89(3):405-67
references: 14894446
references: J Neurophysiol. 1997 Mar;77(3):1614-23
references: 9084624
references: Brain Struct Funct. 2009 Sep;213(4-5):365-6
references: 19198876
references: Int Rev Neurobiol. 1966;9:1-43
references: 5337726
references: Am J Vet Res. 1968 Dec;29(12):2351-64
references: 5749118
references: Brain Res. 2004 Mar 19;1001(1-2):125-32
references: 14972661
references: Science. 1989 Jul 14;245(4914):192-4
references: 2749258
references: J Physiol. 2003 Jul 1;550(Pt 1):135-47
references: 12879864
references: Mol Cell Neurosci. 2004 Jun;26(2):241-50
references: 15207849
references: J Neurosci Methods. 1991 Apr;37(2):173-82
references: 1715499
references: Nat Protoc. 2006;1(3):1439-45
references: 17406432
references: Neurosci Res. 2003 Aug;46(4):523-32
references: 12871774
references: Nature. 2006 Mar 16;440(7082):352-7
references: 16541076
references: Curr Drug Targets CNS Neurol Disord. 2005 Aug;4(4):435-52
references: 16101559
references: Neurochem Int. 2004 Jul;45(1):117-27
references: 15082229
references: J Virol. 2004 Feb;78(3):1212-8
references: 14722276
references: Parasitology. 2005 Dec;131(Pt 6):797-804
references: 16336733
references: J Parasitol. 2005 Apr;91(2):307-15
references: 15986605
references: J Exp Zool. 1947 Oct;106(1):85-107
references: 20268087
references: J Neurosci. 2004 Mar 31;24(13):3370-8
references: 15056716
references: J Virol. 2005 Sep;79(18):11716-23
references: 16140749
references: Epilepsia. 2004 Apr;45(4):322-32
references: 15030494
references: J Neurochem. 2003 Apr;85(1):214-23
references: 12641743
references: J Neuropathol Exp Neurol. 2004 Jun;63(6):610-7
references: 15217089
references: Neurosci Lett. 1996 Jun 28;211(3):203-6
references: 8817576
references: Exp Neurol. 2011 Apr;228(2):222-31
references: 21241693
references: J Comp Neurol. 1984 Sep 20;228(3):432-46
references: 6148364
references: Brain Res. 2000 Jun 9;867(1-2):62-9
references: 10837798
references: Neuroscience. 1984 Apr;11(4):751-60
references: 6330608
references: Neurobiol Dis. 2002 Dec;11(3):355-68
references: 12586546