Gill membrane remodeling with soft-water acclimation in zebrafish (Danio rerio)

Little is known regarding the ionoregulatory abilities of zebrafish exposed to soft water despite the popularity of this model organism for physiology and aquatic toxicology. We examined genomic and nongenomic changes to gills of zebrafish as they were progressively acclimated from moderately hard freshwater to typical soft water over 7 days and held in soft water for another 7 days. Gills were sampled daily and mRNA expression levels of gill Na(+)-K(+)-ATPase (NKA) alpha1a subunit, epithelium calcium channel (ECaC), carbonic anhydrase-1 and 2 (CA-1, CA-2), Na(+)/H(+) exchanger (NHE-2), V-type proton (H(+))-ATPase, and copper transport protein (CTR-1) were quantified by real-time PCR. Changes in enzyme activities of gill NKA were determined and protein levels of NKA and ECaC were quantified by Western blotting. Levels of mRNA for ECaC increased fourfold after day 6, with an associated increase in ECaC protein levels after 1 wk in soft water. CA-1 and CA-2 exhibited a 1.5- and 6-fold increase in gene expression on days 6 and 5, respectively. Likewise, there was a fivefold increase in NHE-2 expression after day 6. Surprisingly, CTR-1 mRNA showed a large transient increase (over threefold) on day 6, while H(+)-ATPase mRNA did not change. These data demonstrate a high degree of phenotypic plasticity in zebrafish gills exposed to an ion-poor environment. This not only enhances our understanding of ionoregulatory processes in fish but also highlights the need for proper experimental design for studies involving preacclimation to soft water (e.g., metal toxicity).