Transcriptional analysis of rat piriform cortex following exposure to the organophosphonate anticholinesterase sarin and induction of seizures

Journal of Neuroinflammation

James F Dillman²  James L Meyerhoff³  Christopher L Robison¹  Lucille A Lumley¹  Kimberly D Spradling² 

[1] Neurobehavioral Toxicology Branch, US Army Medical Research Institute of Chemical Defense (USAMRICD), 3100 Ricketts Point Road, Aberdeen Proving Ground, MD 21010-5400, USA;Cell and Molecular Biology Branch, US Army Medical Research Institute of Chemical Defense (USAMRICD), 3100 Ricketts Point Road, Aberdeen Proving Ground, MD 21010-5400, USA;US Army Center for Environmental Health Research, 568 Doughten Drive, Fort Detrick, MD 21702-5010, USA

关键词: Transcriptomics;    Microarray;    Cytokine;    Neuroinflammation;    Seizure;    Piriform Cortex;    Sarin;    Organophosphate;    Chemical Warfare;    Nerve Agent;   

Others  :  1220801
DOI  :  10.1186/1742-2094-8-83

 received in 2011-04-27, accepted in 2011-07-21,  发布年份 2011

【 摘 要 】

Background

Organophosphorus nerve agents irreversibly inhibit acetylcholinesterase, causing a toxic buildup of acetylcholine at muscarinic and nicotinic receptors. Current medical countermeasures to nerve agent intoxication increase survival if administered within a short period of time following exposure but may not fully prevent neurological damage. Therefore, there is a need to discover drug treatments that are effective when administered after the onset of seizures and secondary responses that lead to brain injury.

Methods

To determine potential therapeutic targets for such treatments, we analyzed gene expression changes in the rat piriform cortex following sarin (O-isopropyl methylphosphonofluoridate)-induced seizure. Male Sprague-Dawley rats were challenged with 1 × LD₅₀ sarin and subsequently treated with atropine sulfate, 2-pyridine aldoxime methylchloride (2-PAM), and the anticonvulsant diazepam. Control animals received an equivalent volume of vehicle and drug treatments. The piriform cortex, a brain region particularly sensitive to neural damage from sarin-induced seizures, was extracted at 0.25, 1, 3, 6, and 24 h after seizure onset, and total RNA was processed for microarray analysis. Principal component analysis identified sarin-induced seizure occurrence and time point following seizure onset as major sources of variability within the dataset. Based on these variables, the dataset was filtered and analysis of variance was used to determine genes significantly changed in seizing animals at each time point. The calculated p-value and geometric fold change for each probeset identifier were subsequently used for gene ontology analysis to identify canonical pathways, biological functions, and networks of genes significantly affected by sarin-induced seizure over the 24-h time course.

Results

A multitude of biological functions and pathways were identified as being significantly altered following sarin-induced seizure. Inflammatory response and signaling pathways associated with inflammation were among the most significantly altered across the five time points examined.

Conclusions

This analysis of gene expression changes in the rat brain following sarin-induced seizure and the molecular pathways involved in sarin-induced neurodegeneration will facilitate the identification of potential therapeutic targets for the development of effective neuroprotectants to treat nerve agent exposure.

【 授权许可】

   
2011 Spradling et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

Figure 8.

Figure 9.

Figure 10.

Figure 11.

Figure 12.

【 参考文献 】

• [1]Schmaltz F: Neurosciences and research on chemical weapons of mass destruction in Nazi Germany. J Hist Neurosci 2006, 15:186-209.
• [2]Brown MA, Brix KA: Review of health consequences from high-, intermediate and low-level exposure to organophosphorus nerve agents. J Appl Toxicol 1998, 18:393-408.
• [3]Hoffman A, Eisenkraft A, Finkelstein A, Schein O, Rotman E, Dushnitsky T: A decade after the Tokyo sarin attack: a review of neurological follow-up of the victims. Mil Med 2007, 172(6):607-610.
• [4]Aroniadou-Anderjaska V, Figueiredo TH, Apland JP, Qashu F, Braga MFM: Primary brain targets of nerve agents: the role of the amygdala in comparison to the hippocampus. Neurotoxicology 2009, 30:772-776.
• [5]Loh Y, Swanberg MM, Ingram MV, Newmark J: Case report: long-term cognitive sequelae of sarin exposure. Neurotoxicology 2010, 31:244-246.
• [6]McDonough JH Jr, Shih TM: Neuropharmacological mechanisms of nerve agent-induced seizure and neuropathology. Neurosci Biobehav Rev 1997, 21:559-579.
• [7]Bajgar J: Complex view on poisoning with nerve agents and organophosphates. Acta Medica (Hradec Králové) 2005, 48(1):3-21.
• [8]Filbert M, Levine E, Ballough G: Neuroprotection for nerve agent-induced brain damage by blocking delayed calcium overload: a review. J Med Chem Biol Radiol Def 2005, 3:1-21.
• [9]Myhrer T, Enger S, Aas P: Efficacy of immediate and subsequent therapies against soman-induced seizures and lethality in rats. Basic Clin Pharmacol Toxicol 2006, 98:184-191.
• [10]Myhrer T: Neuronal structures involved in the induction and propagation of seizures caused by nerve agents: implications for medical treatment. Toxicology 2007, 239(1-2):1-14.
• [11]Angoa-Pérez M, Kreipke CW, Thomas DM, Van Shura KE, Lyman M, McDonough JH, Kuhn DM: Soman increases neuronal COX-2 levels: possible link between seizures and protracted neuronal damage. Neurotoxicology 2010, 31:738-746.
• [12]Bajgar J, Ševelová L, Krejčová G, Fusek J, Vachek J, Kassa J, Herink J: Biochemical and behavioral effects of soman vapors in low concentrations. Inhal Toxicol 2004, 16:497-507.
• [13]Myhrer T, Enger S, Aas P: Anticonvulsant effects of damage to structures involved in seizure induction in rats exposed to soman. Neurotoxicology 2007, 28:819-828.
• [14]Joosen MJA, Jousma E, van den Boom TM, Kuijpers WC, Smit AB, Lucassen PJ, van Helden HPM: Long-term cognitive deficits accompanied by reduced neurogenesis after soman poisoning. Neurotoxicology 2009, 30:72-80.
• [15]Baille V, Clarke PGH, Brochier G, Dorandeu F, Verna JM, Four E, Lallement G, Carpentier P: Soman-induced convulsions: the neuropathology revisited. Toxicology 2005, 215:1-24.
• [16]Scremin OU, Shih TM, Huynh L, Roch M, Booth R, Jenden DJ: Delayed neurologic and behavioral effects of subtoxic doses of cholinesterase inhibitors. J Pharmacol Exp Ther 2003, 304(3):1111-1119.
• [17]Afshari CA: Perspective: microarray technology, seeing more than spots. Endocrinology 2002, 143(6):1983-1989.
• [18]Barlow T, Battershill J, Jeffery BR, Pollitt FD, Tahourdin CSM: Report of a symposium on the use of genomics and proteomics in toxicology. Mutagenesis 2003, 18(3):311-317.
• [19]Blanton JL, D'Ambrozio JA, Sistrunk JE, Midboe EG: Global changes in the expression patterns of RNA isolated from the hippocampus and cortex of VX exposed mice. J Biochem Mol Toxicol 2004, 18(3):115-123.
• [20]Dillman JF III, Phillips CS, Dorsch LM, Croxton MD, Hege AI, Sylvester AJ, Moran TS, Sciuto AM: Genomic analysis of rodent pulmonary tissue following bis-(2-chloroethyl) sulfide exposure. Chem Res Toxicol 2005, 18:28-34.
• [21]Dillman JF III, Hege AI, Phillips CS, Orzolek LD, Sylvester AJ, Bossone C, Henemyre-Harris C, Kiser RC, Choi YW, Schlager JJ, Sabourin CL: Microarray analysis of mouse ear tissue exposed to bis-(2-chloroethyl) sulfide: gene expression profiles correlate with treatment efficacy and an established clinical endpoint. J Pharmacol Exp Ther 2006, 317:76-87.
• [22]Damodaran TV, Greenfield ST, Patel AG, Dressman HK, Lin SK, Abou-Donia MB: Toxicogenomic studies of the rat brain at an early time point following acute sarin exposure. Neurochem Res 2006, 31:367-381.
• [23]Damodaran TV, Patel AG, Greenfield ST, Dressman HK, Lin SM, Abou-Donia MB: Gene expression profiles of the rat brain both immediately and 3 months following acute sarin exposure. Biochem Pharmacol 2006, 71:497-520.
• [24]Dillman JF III, Phillips CS, Kniffin DM, Tompkins CP, Hamilton TA, Kan RK: Gene expression profiling of rat hippocampus following exposure to the acetylcholinesterase inhibitor soman. Chem Res Toxicol 2009, 22(4):633-638.
• [25]Zimmer LA, Ennis M, Shipley MT: Soman-induced seizures rapidly activate astrocytes and microglia in discrete brain regions. J Comp Neurol 1997, 378:482-492.
• [26]Svensson I, Waara L, Johansson L, Bucht A, Cassel G: Soman-induced interleukin-1β mRNA and protein in rat brain. Neurotoxicology 2001, 22:355-362.
• [27]Williams AJ, Berti R, Yao C, Price RA, Velarde LC, Koplovitz I, Schultz SM, Tortella FC, Dave JR: Central neuro-inflammatory gene response following soman exposure in the rat. Neurosci Lett 2003, 349:147-150.
• [28]Svensson I, Waara L, Cassel G: Effects of HI 6, diazepam and atropine on soman-induced IL-1β protein in rat brain. Neurotoxicology 2005, 26:173-181.
• [29]Chapman S, Kadar T, Gilat E: Seizure duration following sarin exposure affects neuro-inflammatory markers in the rat brain. Neurotoxicology 2006, 27:277-283.
• [30]Dhote F, Peinnequin A, Carpentier P, Baille V, Delacour C, Foquin A, Lallement G, Dorandeu F: Prolonged inflammatory gene response following soman-induced seizures in mice. Toxicology 2007, 238:166-176.
• [31]Johnson EA, Kan RK: The acute phase response and soman-induced status epilepticus: temporal and regional changes in rat brain cytokine concentrations. J Neuroinflammation 2010, 7(40):1-9.
• [32]Racine RJ: Modification of seizure activity by electrical stimulation. II. Motor seizure. Electroencephalogr Clin Neurophysiol 1972, 32(3):281-294.
• [33]Ellman GL, Courtney DK, Andres V, Featherstone RM: A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 1961, 7:88-95.
• [34]Irizarry RA, Bolstad BM, Collin F, Cope LM, Hobbs B, Speed TP: Summaries of Affymetrix GeneChip probe level data. Nucleic Acids Res 2003, 31(4):1-8.
• [35]Pearson K: On lines and planes of closest fit to systems of points in space. Philos Mag 1901, 2:559-572.
• [36]Carpentier P, Foquin A, Rondouin G, LernerNatoli M, deGroot DMG, Lallement G: Effects of atropine sulphate on seizure activity and brain damage produced by soman in guinea pigs: ECoG correlates of neuropathology. Neurotoxicology 2000, 4:521-540.
• [37]Damodaran T, Jones KH, Patel AG, Abou-Donia MB: Sarin (nerve agent GB)-induced differential expression of mRNA coding for the acetylcholinesterase gene in the rat central nervous system. Biochem Pharmacol 2003, 65:2041-2047.
• [38]Bansal I, Waghmare CK, Anand T, Gupta AK, Bhattacharya BK: Differential mRNA expression of acetylcholinesterase in the central nervous system of rats with acute and chronic exposure of sarin & physostigmine. J Appl Toxicol 2009, 29:386-394.
• [39]Shih TM: Time course effects of soman on acetylcholine and choline levels in six discrete areas of the rat brain. Psychopharmacology (Berl) 1982, 78(2):170-5.
• [40]Flynn CJ, Wecker L: Elevated choline levels in brain: a non-cholinergic component of organophosphate toxicity. Biochem Pharmacol 1986, 35(18):3115-3121.
• [41]Berridge MJ: Unlocking the secrets of cell signaling. Annu Rev Physiol 2005, 67:1-21.
• [42]Fauvelle F, Dorandeu F, Carpentier P, Foquin A, Rabeson H, Graveron-Demilly D, Arvers P, Testylier G: Changes in mouse brain metabolism following a convulsive dose of soman: a proton HRMAS NMR study. Toxicology 2010, 267(1-3):99-111.
• [43]Vezzani A: Inflammation and epilepsy. Curr Rev Basic Sci 2005, 5(1):1-6.
• [44]Choi J, Koh S: Role of brain inflammation in epileptogenesis. Yonsei Med J 2008, 49(1):1-18.
• [45]Minami M, Kuraishi T, Satoh M: Effects of kainic acid on messenger RNA levels of IL-1 beta, IL-6, TNF alpha and LIF in the rat brain. Biochem Biophys Res Comm 1991, 176:593-598.
• [46]De Simoni MG, Perego C, Ravizza T, Moneta D, Conti M, Marchesi F, De Luigi A, Garattini S, Vezzani A: Inflammatory cytokines and related genes are induced in the rat hippocampus by limbic status epilepticus. Eur J Neurosci 2000, 12:2623-2633.
• [47]Oprica M, Riksson C, Schultzberg M: Inflammatory mechanisms associated with brain damage induced by kainic acid with special reference to the interleukin-1 system. J Cell Mol Med 2003, 7:127-140.
• [48]Duysen EG, Li B, Xie W, Schopfer LM, Anderson RS, Broomfield CA, Lockridge O: Evidence for nonacetylcholinesterase targets of organophosphorus nerve agent: supersensitivity of acetylcholinesterase knockout mouse to VX lethality. J Pharmacol Exp Ther 2001, 299:528-535.