Involvement of purinergic system in inflammation and toxicity induced by copper in zebrafish larvae.
The use of zebrafish (Danio rerio) is increasing as an intermediate preclinical model, to prioritize drug candidates for mammalian testing. As the immune system of the zebrafish is quite similar to that of mammals, models of inflammation are being developed for the screening of new drugs. The characterization of these models is crucial for studies that seek for mechanisms of action and specific pharmacological targets. It is well known that copper is a metal that induces damage and cell migration to hair cells of lateral line of zebrafish. Extracellular nucleotides/nucleosides, as ATP and adenosine (ADO), act as endogenous signaling molecules during tissue damage by exerting effects on inflammatory and immune responses. The present study aimed to characterize the inflammatory status, and to investigate the involvement of the purinergic system in copper-induced inflammation in zebrafish larvae. Fishes of 7 days post-fertilization were exposed to 10 μM of copper for a period of 24 h. The grade of oxidative stress, inflammatory status, copper uptake, the activity and the gene expression of the enzymes responsible for controlling the levels of nucleotides and adenosine were evaluated. Due to the copper accumulation in zebrafish larvae tissues, the damage and oxidative stress were exacerbated over time, resulting in an inflammatory process involving IL-1β, TNF-α, COX-2 and PGE2. Within the purinergic system, the mechanisms that control the ADO levels were the most involved, mainly the reactions performed by the isoenzyme ADA 2. In conclusion, our data shed new lights on the mechanisms related to copper-induced inflammation in zebrafish larvae.
Instituto de Toxicologia e Farmacologia, Pontifícia Universidade Católica do Rio Grande do Sul, Porto Alegre, CEP 90619-900, Brazil; Programa de Pós-Graduação em Medicina: Ciências Médicas, Universidade Federal do Rio Grande do Sul, Porto Alegre, CEP 90035-003, Brazil. Electronic address: carlos.leite@pucrs.br.
Carlos Eduardo
CE
Leite
Lucas de Oliveira
Lde O
Maboni
Fernanda Fernandes
FF
Cruz
Denis Broock
DB
Rosemberg
Fernanda Francine
FF
Zimmermann
Talita Carneiro Brandão
TC
Pereira
Maurício Reis
MR
Bogo
Carla Denise
CD
Bonan
Maria Martha
MM
Campos
Fernanda Bueno
FB
Morrone
Ana Maria Oliveira
AM
Battastini
D000818:Animals; D003300:Copper / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D007249:Inflammation / Q000139:chemically induced / Q000378:metabolism / Q000503:physiopathology; D007814:Larva / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects* / Q000502:physiology; D011684:Purine Nucleosides / Q000502:physiology*; D011685:Purine Nucleotides / Q000502:physiology*; D015027:Zebrafish / Q000196:embryology
D011684:Purine Nucleosides; D011685:Purine Nucleotides; D003300:Copper
ADA; ADO; ADP; AMP; ATP; Adenosine; CAT; COX-2; Copper; DAD; Diode array detector; E-NTPDase; Ecto-5′-NT; GSH; HPLC-FLD; ICP-MS; IL-10; IL-1β; INO; Inductively coupled plasma mass spectrometry; Inflammation; Inosine; Interleukin 10; Interleukin 1β; MPO; Myeloperoxidase; NO; Nitric oxide; PGE(2); Prostaglandin E(2); Purinergic system; SOD; Superoxide dismutase; TNF-α; Tumor necrosis factor α; UHPLC; Ultra high performance liquid chromatography coupled with mass spectrometry; Zebrafish; adenosine; adenosine deaminase; adenosine diphosphate; adenosine monophosphate; adenosine triphosphate; catalase; cyclooxygenase 2; ecto-5′-nucleotidase; ecto-nucleoside-trifosfo difosfoidrolases; high performance liquid chromatography with fluorescence detector; reduced glutathione