Immunohistochemical Approach to Study Cylindrospermopsin Distribution in Tilapia (Oreochromis niloticus) under Different Exposure Conditions

Toxins

Remedios Guzmán-Guillén¹  Daniel Gutiérrez-Praena¹  Mar໚ de los Ángeles Risalde⁴  Rosario Moyano²  Ana Isabel Prieto¹  Silvia Pichardo¹  Ángeles Jos¹  Vitor Vasconcelos³ 

[1] Area of Toxicology, Faculty of Pharmacy, University of Seville, Profesor García González 2, Seville 41012, Spain; E-Mails:;Department of Pharmacology, Toxicology and Legal and Forensic Medicine, University of Córdoba, Campus de Rabanales, Carretera Madrid-Cádiz s/n, Córdoba 14071, Spain; E-Mail:;Laboratory of Ecotoxicology, Genomics and Evolution, Interdisciplinary Center of Marine and Environmental Research—CIIMAR/CIMAR, University of Porto, Rua dos Bragas 289, Porto 4050-123, Portugal; E-Mail:;Department of Anatomy and Comparative Pathology and Anatomy, University of Córdoba, Campus de Rabanales, Carretera Madrid-Cádiz s/n, Córdoba 14071, Spain; E-Mail:

关键词: Cylindrospermopsin;    Aphanizomenon ovalisporum;    Oreochromis niloticus;    immunohistochemistry;    distribution;    cyanobacteria;    cyanotoxin;   

DOI  :  10.3390/toxins6010283

来源: mdpi

【 摘 要 】

Cylindrospermopsin (CYN) is a cytotoxic cyanotoxin produced by several species of freshwater cyanobacteria (i.e., Aphanizomenon ovalisporum). CYN is a tricyclic alkaloid combined with a guanidine moiety. It is well known that CYN inhibits both protein and glutathione synthesis, and also induces genotoxicity and the alteration of different oxidative stress biomarkers. Although the liver and kidney appear to be the main target organs for this toxin based on previous studies, CYN also affects other organs. In the present study, we studied the distribution of CYN in fish (Oreochromis niloticus) under two different exposure scenarios using immunohistochemical (IHC) techniques. In the first method, fish were exposed acutely by intraperitoneal injection or by gavage to 200 µg pure CYN/Kg body weight (bw), and euthanized after 24 h or five days of exposure. In the second method, fish were exposed by immersion to lyophilized A. ovalisporum CYN-producing cells using two concentration levels (10 or 100 µg/L) for two different exposure times (7 or 14 days). The IHC was carried out in liver, kidney, intestine, and gills of fish. Results demonstrated a similar pattern of CYN distribution in both experimental methods. The organ that presented the most immunopositive results was the liver, followed by the kidney, intestine, and gills. Moreover, the immunolabeling signal intensified with increasing time in both assays, confirming the delayed toxicity of CYN, and also with the increment of the dose, as it is shown in the sub-chronic assay. Thus, IHC is shown to be a valuable technique to study CYN distribution in these organisms.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.