Name: screening assay for oxidative stress in a feline astrocyte cell line, g355-5
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1. Colture cellulari di astrociti felino e il trattamento con H 2 O 2 <ol><li> Linea cellulare cultura G355-5 in un centimetro 75 2 pallone con i media DMEM più nutrienti a 37 ° C e 5% di CO 2 fino al 60% confluenti (~ 5 giorni). Sostituire i media ogni 1-2 giorni o secondo necessità per mantenere una sana cultura. </li><li> Rimuovere i supporti e aggiungere 2 ml di tripsina 0,25%. Sollevare le cellule pipettando per 45 s - 1 minuti a temperatura ambiente (RT). </li><li> ...

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	<li>Bukrinsky, M. I., Nottet, H. S., Schmidtmayerova, H. L., Dubrovsky, H., Flanagan, C. R., Mullins, M. E., Lipton, S. A., Gendelman, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+nitric+oxide+synthase+activity+in+human+immunodeficiency+virus+type+1+(HIV-1)-infected+monocytes:+implications+for+HIV-associated+neurological+disease.">Regulation of nitric oxide synthase activity in human immunodeficiency virus type 1 (HIV-1)-infected monocytes: implications for HIV-associated neurological disease.</a> J Exp Med. 181, 735-745 (1995).</li><li>Cadet, J. L., Ali, S., Epstein, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Involvement+of+oxygen-based+radicals+in+methamphetamine-induced+neurotoxicity:+evidence+from+the+use+of+CuZnSOD+transgenic+mice.">Involvement of oxygen-based radicals in methamphetamine-induced neurotoxicity: evidence from the use of CuZnSOD transgenic mice.</a> Ann N Y Acad Sci. 738, 388-3891 (1994).</li><li>Cathcart, R., Schwiers, E., Ames, B. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+picomole+levels+of+hydroperoxides+using+a+fluorescent+dichlorofluorescein+assay.">Detection of picomole levels of hydroperoxides using a fluorescent dichlorofluorescein assay.</a> Anal Biochem. 134, 111-116 (1983).</li><li>Chao, C. C., Hu, S., Peterson, P. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glia:+the+not+so+innocent+bystanders.">Glia: the not so innocent bystanders.</a> J Neurovirol. 2, 234-239 (1996).</li><li>Cooper, A. J., Kristal, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+roles+of+glutathione+in+the+central+nervous+system.">Multiple roles of glutathione in the central nervous system.</a> Biol Chem. 378, 793-802 (1997).</li><li>Cubells, J. F., Rayport, S., Rajendran, G., Sulzer, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methamphetamine+neurotoxicity+involves+vacuolation+of+endocytic+organelles+and+dopamine-dependent+intracellular+oxidative+stress.">Methamphetamine neurotoxicity involves vacuolation of endocytic organelles and dopamine-dependent intracellular oxidative stress.</a> J Neurosci. 14, 2260-2271 (1994).</li><li>Dawson, V. L., Dawson, T. M., Uhl, G. R., Snyder, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+immunodeficiency+virus+type+1+coat+protein+neurotoxicity+mediated+by+nitric+oxide+in+primary+cortical+cultures.">Human immunodeficiency virus type 1 coat protein neurotoxicity mediated by nitric oxide in primary cortical cultures.</a> Proc Natl Acad Sci U S A. 90, 3256-329 (1993).</li><li>Desagher, S., Glowinski, J., Premont, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Astrocytes+protect+neurons+from+hydrogen+peroxide+toxicity.">Astrocytes protect neurons from hydrogen peroxide toxicity.</a> J Neurosci. 16, 2553-2562 (1996).</li><li>Dewhurst, S., Gelbard, H. A., Fine, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuropathogenesis+of+AIDS.">Neuropathogenesis of AIDS.</a> Mol Med Today. 2, 16-23 (1996).</li><li>Dringen, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolism+and+functions+of+glutathione+in+brain.">Metabolism and functions of glutathione in brain.</a> Prog Neurobiol. 62, 649-671 (2000).</li><li>Dringen, R., Hamprecht, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glutathione+restoration+as+indicator+for+cellular+metabolism+of+astroglial+cells.">Glutathione restoration as indicator for cellular metabolism of astroglial cells.</a> Dev Neurosci. 20, 401-407 (1998).</li><li>Dringen, R., Pfeiffer, B., Hamprecht, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+the+Antioxidant+Glutathione+in+Neurons:+Supply+by+Astrocytes+of+CysGly+as+Precursor+for+Neuronal+Glutathione.">Synthesis of the Antioxidant Glutathione in Neurons: Supply by Astrocytes of CysGly as Precursor for Neuronal Glutathione.</a> J Neurosci. 19, 562-569 (1999).</li><li>Everall, I. P. H. u. d. s. o. n., L, R. W. K. e. r. w. i. n. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+absolute+levels+of+ascorbic+acid+and+unaltered+vasoactive+intestinal+polypeptide+receptor+binding+in+the+frontal+cortex+in+acquired+immunodeficiency+syndrome.">Decreased absolute levels of ascorbic acid and unaltered vasoactive intestinal polypeptide receptor binding in the frontal cortex in acquired immunodeficiency syndrome.</a> Neurosci Lett. 224, 119-1122 (1997).</li><li>Gabriel, C., Camins, A., Sureda, F. X., Aquirre, L., Escubedo, E., Pallas, M., Camarasa, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+nitric+oxide+generation+in+mammalian+neurons+using+dichlorofluorescin+diacetate+and+flow+cytometry.">Determination of nitric oxide generation in mammalian neurons using dichlorofluorescin diacetate and flow cytometry.</a> J Pharmacol Toxicol Methods. 38, 93-938 (1997).</li><li>Gendelman, H. E., Genis, P., Jett, M., Zhai, Q. H., Nottet, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+experimental+model+system+for+HIV-1-induced+brain+injury.">An experimental model system for HIV-1-induced brain injury.</a> Adv Neuroimmunol. 4, 189-1893 (1994).</li><li>Hayman, M., Arbuthnott, G., Harkiss, G., Brace, H., Filippi, P., Philippon, V., Thomson, D., Vigne, R., Wright, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurotoxicity+of+peptide+analogues+of+the+transactivating+protein+tat+from+Maedi-Visna+virus+and+human+immunodeficiency+virus.">Neurotoxicity of peptide analogues of the transactivating protein tat from Maedi-Visna virus and human immunodeficiency virus.</a> Neuroscience. 53, 1-6 (1993).</li><li>Hirata, H., Ladenheim, B., Rothman, R. B., Epstein, C., Cadet, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methamphetamine-induced+serotonin+neurotoxicity+is+mediated+by+superoxide+radicals.">Methamphetamine-induced serotonin neurotoxicity is mediated by superoxide radicals.</a> Brain Res. , 677-6345 (1995).</li><li>Koka, P., He, K., Zack, J. A., Kitchen, S., Peacock, W., Fried, I., Tran, T., Yashar, S. S., Merrill, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+immunodeficiency+virus+1+envelope+proteins+induce+interleukin+1,+tumor+necrosis+factor+alpha,+and+nitric+oxide+in+glial+cultures+derived+from+fetal,+neonatal,+and+adult+human+brain.">Human immunodeficiency virus 1 envelope proteins induce interleukin 1, tumor necrosis factor alpha, and nitric oxide in glial cultures derived from fetal, neonatal, and adult human brain.</a> J Exp Med. 182, 941-951 (1995).</li><li>Kong, L. Y., Wilson, B. C., McMillian, M. K., Bing, G., Hudson, P. M., Hong, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+the+HIV-1+envelope+protein+gp120+on+the+production+of+nitric+oxide+and+proinflammatory+cytokines+in+mixed+glial+cell+cultures.">The effects of the HIV-1 envelope protein gp120 on the production of nitric oxide and proinflammatory cytokines in mixed glial cell cultures.</a> Cell Immunol. 172, 77-83 (1996).</li><li>Koutsilieri, E., Gotz, M. E., Sopper, S., Sauer, U., Demuth, M., ter Meulen, V., Riederer, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+glutathione+and+cell+toxicity+following+exposure+to+neurotropic+substances+and+human+immunodeficiency+virus-1+in+vitro.">Regulation of glutathione and cell toxicity following exposure to neurotropic substances and human immunodeficiency virus-1 in vitro.</a> J Neurovirol. 3, 342-349 (1997).</li><li>Lipton, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Similarity+of+neuronal+cell+injury+and+death+in+AIDS+dementia+and+focal+cerebral+ischemia:+potential+treatment+with+NMDA+open-channel+blockers+and+nitric+oxide-related+species.">Similarity of neuronal cell injury and death in AIDS dementia and focal cerebral ischemia: potential treatment with NMDA open-channel blockers and nitric oxide-related species.</a> Brain Pathol. 6, 507-517 (1996).</li><li>Lipton, S. A., Yeh, M., Dreyer, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Update+on+current+models+of+HIV-related+neuronal+injury:+platelet-activating+factor,+arachidonic+acid+and+nitric+oxide.">Update on current models of HIV-related neuronal injury: platelet-activating factor, arachidonic acid and nitric oxide.</a> Adv Neuroimmunol. 4, 181-188 (1994).</li><li>Mills, E. M., Takeda, K., Yu, Z. X., Ferrans, V., Katagiri, Y., Jiang, H., Lavigne, M. C., Leto, T. L., Guroff, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nerve+growth+factor+treatment+prevents+the+increase+in+superoxide+produced+by+epidermal+growth+factor+in+PC12+cells.">Nerve growth factor treatment prevents the increase in superoxide produced by epidermal growth factor in PC12 cells.</a> J Biol Chem. 273, 22165-22168 (1998).</li><li>Peuchen, S., Duchen, M. R., Clark, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+the+glutathione+redox+state+in+adult+astrocytes.">Modulation of the glutathione redox state in adult astrocytes.</a> Biochem Soc Trans. 24, 449S-449S (1996).</li><li>Possel, H., Noack, H., Augustin, W., Keilhoff, G., Wolf, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=2,7-Dihydrodichlorofluorescein+diacetate+as+a+fluorescent+marker+for+peroxynitrite+formation.">2,7-Dihydrodichlorofluorescein diacetate as a fluorescent marker for peroxynitrite formation.</a> FEBS Lett. 416, 175-178 (1997).</li><li>Sagara, J., Makino, N., Bannai, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glutathione+efflux+from+cultured+astrocytes.">Glutathione efflux from cultured astrocytes.</a> J Neurochem. 66, 1876-1881 (1996).</li><li>Stefano, G. B., Smith, E. M., Paemen, L. R., Hughes, T. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HIV+gp120+associated+neurological+deficits:+a+potential+role+for+nitric+oxide+and+other+signal+molecules.">HIV gp120 associated neurological deficits: a potential role for nitric oxide and other signal molecules.</a> Advances in Neuroimmunology. 3, 47-57 (1993).</li><li>Sundaresan, M., Yu, Z. X., Ferrans, V. J., Irani, K., Finkel, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Requirement+for+generation+of+H2O2+for+platelet-derived+growth+factor+signal+transduction.">Requirement for generation of H2O2 for platelet-derived growth factor signal transduction.</a> Science. 270, 296-299 (1995).</li><li>Wang, X. F., Cynader, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Astrocytes+provide+cysteine+to+neurons+by+releasing+glutathione.">Astrocytes provide cysteine to neurons by releasing glutathione.</a> J Neurochem. 74, 1434-1442 (2000).</li><li>Wilson, J. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antioxidant+defense+of+the+brain:+a+role+for+astrocytes.">Antioxidant defense of the brain: a role for astrocytes.</a> Can J Physiol Pharmacol. 75, 1149-1163 (1997).</li><li>Zenger, E., Collisson, E. W., Barhoumi, R., Burghardt, R. C., Danave, I. R., Tiffany-Castiglioni, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser+cytometric+analysis+of+FIV-induced+injury+in+astroglia.">Laser cytometric analysis of FIV-induced injury in astroglia.</a> Glia. 13, 92-100 (1995).</li></ol>

Un meccanismo spesso suggerito di virus danno neuronale indotto è lo stress ossidativo 1, 4, 7, 9, 13, 15, 16, 18-22, 27, 31. In sostanza, si propone che attraverso l&#39;esposizione virale della glia (astrociti e microglia) radicali rilasciano ossigeno reattive come, idrossido, anione superossido, ossido nitrico, e / o perossido di idrogeno, che sono tossici per i neuroni. Allo stesso modo, lo stress ossidativo è stato anche suggerito come un importante meccanismo patologico in metanfetamine neurotossicit?...

<table border="1"><tbody><tr><th> Nome del reagente </th><th> Azienda </th><th> Numero di catalogo </th><th> Commenti (opzionale) </th></tr><tr><td> DMEM </td><td> Cellgro </td><td> 15-013-CV </td><td></td></tr><tr><td> MEM Vitamine </td><td> Cellgro </td><td> 25-020-cl </td><td> 1% (v / v) </td></tr><tr><td> L-glutammina </td><td> Hyclone </td><td> 17-605E </td><td> 1% (v / v) </td></tr><tr><td> Non amminoacidi essenziali </td><td> Hyclone </td><td> 25-025-cl </td><td> 1% (v / v) </td></tr><tr><td> FBS </td><td> Hyclone </td><td> SH30071.03 </td><td> 20% (v / v) </td></tr><tr><td> Aminoacidi essenziali </td><td> Cellgro </td><td> 25-030-cl </td><td> 0,2% (v / v) </td></tr><tr><td> 500ml vuoto sistema di filtrazione </td><td> VWR </td><td> 87006-076 </td><td></td></tr><tr><td> Provette da 15 ml </td><td> Falco </td><td> 352097 </td><td></td></tr><tr><td> 75 centimetri due colture di tessuti fiaschi </td><td> Falco </td><td> 353136 </td><td></td></tr><tr><td> 6 e piastra di coltura dei tessuti </td><td> Falco </td><td> 353224 </td><td></td></tr><tr><td> CM-H 2 DCFDA </td><td> Invitrogen </td><td> C6827 </td><td></td></tr><tr><td> Propidio ioduro (PI) </td><td> Sigma </td><td> P4170-10mg </td><td></td></tr><tr><td> Tripsina </td><td> Lonza </td><td> 17-160F </td><td></td></tr><tr><td> H 2 O 2 </td><td> Sigma </td><td> H6520 </td><td></td></tr><tr><td> HyQ antibiotici </td><td> Hyclone </td><td> SV30079.01 </td><td> 0,1% (v / v) </td></tr><tr><td> G355-5 cellule </td><td> ATCC </td><td> CRL-2033 </td><td> Normale cervello felino </td></tr></tbody></table>

screening assay for oxidative stress in a feline astrocyte cell line, g355-5

Un meccanismo spesso suggerito di virus danno neuronale indotto è lo stress ossidativo. Astrociti svolgono un ruolo importante nel controllo dello stress ossidativo del Sistema Nervoso Centrale (SNC). Astrociti aiutare a mantenere un ambiente omeostatico per i neuroni così come proteggere i neuroni da specie reattive dell&#39;ossigeno (ROS). CM-H2DCFDA è una cellula-permeabile indicatore per la presenza di ROS. CM-H 2 DCFDA entra nella cellula come un non-fluorescenti composte, e diventa fluorescente dopo esterasi cellulari rimuovere i gruppi di acetato, e il composto viene ossidato. Il numero di cellule, misurato mediante citometria di flusso, che si trovano ad essere fluorescente verde è l&#39;indicazione del numero di cellule che sono in uno stato ossidativo. CM-H 2 DCFDA è suscettibile di ossidazione da un gran numero di differenti ROS. Questa mancanza di specificità, per i quali ROS possono ossidare CM-H 2 DCFDA, rende questo composto un reggente prezioso per l&#39;utilizzo nelle prime fasi di un&#39;indagine patogenesi, in quanto questo test può essere usato per individuare un ambiente ossidativo cellulare, indipendentemente da quale ossigeno radicale o la combinazione di ROS sono responsabili per le condizioni di cellulare. Una volta stabilito che i ROS sono presenti dall&#39;ossidazione di CM-H 2 DCFDA, poi ulteriori esperimenti possono essere eseguiti per determinare quale ROS o combinazione di Ross sono coinvolti nel processo di patogenesi particolare. I risultati di questo studio dimostrano che con l&#39;aggiunta di perossido di idrogeno, un aumento della CM-H 2 DCFDA fluorescenza è stato rilevato rispetto ai controlli salina, indicando che questo test è un test importante per la rilevazione di un ambiente ossidativo all&#39;interno G355-5 celle, un felina linea cellulare astrociti.

Watch this Scientific Journal Video about Screening Assay for Oxidative Stress in a Feline Astrocyte Cell Line, G355-5 at JoVE.com

Screening Assay for Oxidative Stress in a Feline Astrocyte Cell Line, G355-5

Un metodo di screening per individuare gli ambienti ossidativo cellulare è quello di misurare l&#39;ossidazione delle CM-H2DCFDA. Una volta ossidato all&#39;interno di una cella di CM-H2DCFDA cambiamenti da non fluorescente in un composto fluorescente. Questo cambiamento di fluorescenza viene misurata mediante citometria di flusso e indica il numero di cellule in un ambiente ossidativo.

Test di screening per Stress Ossidativo in una linea felina cella astrociti, G355-5

An often-suggested mechanism of virus induced neuronal damage is oxidative stress. Astrocytes have an important role in controlling oxidative stress of ...

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