title: Indomethacin treatment prior to pentylenetetrazole-induced seizures downregulates the expression of il1b and cox2 and decreases seizure-like behavior in zebrafish larvae.
abstract: BACKGROUND
  It has been demonstrated that the zebrafish model of pentylenetetrazole (PTZ)-evoked seizures and the well-established rodent models of epilepsy are similar pertaining to behavior, electrographic features, and c-fos expression. Although this zebrafish model is suitable for studying seizures, to date, inflammatory response after seizures has not been investigated using this model. Because a relationship between epilepsy and inflammation has been established, in the present study we investigated the transcript levels of the proinflammatory cytokines interleukin-1 beta (il1b) and cyclooxygenase-2 (cox2a and cox2b) after PTZ-induced seizures in the brain of zebrafish 7 days post fertilization. Furthermore, we exposed the fish to the nonsteroidal anti-inflammatory drug indomethacin prior to PTZ, and we measured its effect on seizure latency, number of seizure behaviors, and mRNA expression of il1b, cox2b, and c-fos. We used quantitative real-time PCR to assess the mRNA expression of il1b, cox2a, cox2b, and c-fos, and visual inspection was used to monitor seizure latency and the number of seizure-like behaviors.
  
  
  RESULTS
  We found a short-term upregulation of il1b, and we revealed that cox2b, but not cox2a, was induced after seizures. Indomethacin treatment prior to PTZ-induced seizures downregulated the mRNA expression of il1b, cox2b, and c-fos. Moreover, we observed that in larvae exposed to indomethacin, seizure latency increased and the number of seizure-like behaviors decreased.
  
  
  CONCLUSIONS
  This is the first study showing that il1b and cox-2 transcripts are upregulated following PTZ-induced seizures in zebrafish. In addition, we demonstrated the anticonvulsant effect of indomethacin based on (1) the inhibition of PTZ-induced c-fos transcription, (2) increase in seizure latency, and (3) decrease in the number of seizure-like behaviors. Furthermore, anti-inflammatory effect of indomethacin is clearly demonstrated by the downregulation of the mRNA expression of il1b and cox2b. Our results are supported by previous evidences suggesting that zebrafish is a suitable alternative for studying inflammation, seizures, and the effect of anti-inflammatory compounds on seizure suppression.
authors.affiliation: Laboratory of Zebrafish, Department of Medical Genetics - School of Medical Sciences, University of Campinas (UNICAMP), Tessália Vieira de Camargo, 126 Cidade Universitaria "Zeferino Vaz", Campinas, SP, 13083-887, Brazil. patriciagbarbalho@yahoo.com.br.
authors.forename: Patrícia Gonçalves
authors.initials: PG
authors.lastname: Barbalho
authors.affiliation: Laboratory of Molecular Genetics, Department of Medical Genetics - School of Medical Sciences, University of Campinas (UNICAMP), Tessália Vieira de Camargo, 126 Cidade Universitaria "Zeferino Vaz", Campinas, SP, 13083-887, Brazil. icendes@unicamp.br.
authors.forename: Iscia
authors.initials: I
authors.lastname: Lopes-Cendes
authors.affiliation: Laboratory of Zebrafish, Department of Medical Genetics - School of Medical Sciences, University of Campinas (UNICAMP), Tessália Vieira de Camargo, 126 Cidade Universitaria "Zeferino Vaz", Campinas, SP, 13083-887, Brazil. cmaurer@fcm.unicamp.br.
authors.forename: Claudia Vianna
authors.initials: CV
authors.lastname: Maurer-Morelli
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000008:administration & dosage*; D001921:Brain / Q000187:drug effects / Q000378:metabolism* / Q000503:physiopathology*; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D004195:Disease Models, Animal; D015536:Down-Regulation; D029941:Fish Proteins / Q000378:metabolism; D007213:Indomethacin / Q000008:administration & dosage*; D007249:Inflammation / Q000378:metabolism*; D018836:Inflammation Mediators / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism*; D010433:Pentylenetetrazole; D012333:RNA, Messenger / Q000378:metabolism; D012640:Seizures / Q000139:chemically induced / Q000378:metabolism*; D015027:Zebrafish
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D029941:Fish Proteins; D018836:Inflammation Mediators; D053583:Interleukin-1beta; D012333:RNA, Messenger; D051546:Cyclooxygenase 2; D010433:Pentylenetetrazole; D007213:Indomethacin