Biphasic modulation of neuro- and interrenal steroidogenesis in juvenile African sharptooth catfish (Clarias gariepinus) exposed to waterborne di-(2-ethylhexyl) phthalate.

Receptor (i.e. genomic) and non-receptor (or non-genomic) effects of endocrine toxicology have received limited or almost non-existent attention for tropical species and regions. In the present study, we have evaluated the effects of di-(2-ethylhexyl) phthalate (DEHP) on neuro- and interrenal steroidogenesis of the African catfish (Clarias gariepinus) using molecular, immunochemical and physiological approaches. Juvenile fish (mean weight and length: 5.6±0.6g and 8.2±1.2cm, respectively), were randomly distributed into ten 120L rectangular glass tanks containing 60L of dechlorinated tap water, at 50 fish per exposure group. The fish were exposed to environmentally relevant concentrations of DEHP, consisting of 0 (ethanol solvent control), 10, 100, 200, and 400μg DEHP/L water and performed in two replicates. Brain, liver and head kidney samples were collected at day 3, 7 and 14 after exposure, and analysed for star, p450scc, cyp19a1, cyp17, cyp11β-, 3β-, 17β- and 20β-hsd, and 17β-ohase mRNA expression using real-time PCR. The StAR, P450scc and CYP19 proteins were measured using immunoblotting method, while estradiol-17β (E2) and testosterone (T) were measured in liver homogenate using enzyme immunoassay (EIA). Our data showed a consistent and unique pattern of biphasic effect on star and steroidogenic enzyme genes with increases at low concentration (10μg/L) and thereafter, a concentration-dependent decrease in both the brain and head kidney, that paralleled the expression of StAR, P450scc and CYP19 proteins. Cellular E2 and T levels showed an apparent DEHP concentration-dependent increase at day 14 of exposure. The observed consistency in the current findings and in view of previous reports on contaminants-induced alterations in neuro- and interrenal steroidogenesis, the broader toxicological and endocrine disruptor implication of our data indicate potentials for overt reproductive, metabolic, physiological and general health consequences for the exposed organisms.