Dibutyl phthalate rapidly alters calcium homeostasis in the gills of Danio rerio

Abstract

This study investigates the impacts of exposure to an environment Ca²⁺ challenge and the mechanism of action of dibutyl phthalate (DBP) on Ca²⁺ influx in the gills of Danio rerio. In vitro profile of ⁴⁵Ca²⁺ influx in gills was verified through the basal time-course. Fish were exposed to low, normal and high Ca²⁺ concentrations (0.02, 0.7 and 2 mM) for 12 h. So, gills were morphologically analysed and ex vivo⁴⁵Ca²⁺ influx at 30 and 60 min was determined. For the in vitro studies, gills were treated for 60 min with DBP (1 pM, 1 nM and 1 μM) with/without blockers/activators of ionic channels, Ca²⁺ chelator, inhibitors of ATPases, ionic exchangers and protein kinase C to study the mechanism of DBP-induced ⁴⁵Ca²⁺ influx. Exposure to high environmental Ca²⁺ augmented ⁴⁵Ca²⁺ influx when compared to fish exposed to normal and low Ca²⁺ concentrations. Additionally, histopathological changes were observed in the gills of fish maintained for 12 h in low and high Ca²⁺. In vitro exposure of gills to DBP (1 pM) disturbed Ca²⁺ homeostasis. DBP stimulated ⁴⁵Ca²⁺ influx in gills through the transitory receptor potential vanilloid 1 (TRPV1), and reverse-mode Na⁺/Ca²⁺ exchanger (NCX) activation, protein kinase C and K⁺ channels and sarco/endoplasmic reticulum Ca²⁺-ATPase (SERCA). These data suggest that in vivo short-term exposure of gills to low and high Ca²⁺ leads to ⁴⁵Ca²⁺ influx and histopathological changes. Additionally, the DBP-induced rapid ⁴⁵Ca²⁺ influx is mediated by TRPV1, NCX activation with the involvement of PKC, K⁺-channels and SERCA, thereby altering Ca²⁺ homeostasis.

Keywords: Calcium; Fish; Gills; Histology; Phthalate; Transient receptor potential.