Carbonic anhydrase expression in the branchial ionocytes of rainbow trout

Rainbow trout (Oncorhynchus mykiss) exposed to acid-base challenges activate branchial mechanisms for the excretion of acid-base equivalents. Current models of branchial acid-base excretion in freshwater rainbow trout propose two main ionocyte types; the peanut lectin agglutinin-positive (PNA+) mitochondrion-rich cell or ionocyte is believed to secrete HCO3− in exchange for Cl−, whereas H+ secretion is thought to occur across PNA− ionocytes in exchange for Na+. Both HCO3− and H+ are supplied by intracellular hydration of CO2 catalyzed by cytosolic carbonic anhydrase (CAc). Immunohistochemical approaches revealed that under control conditions, CAc was detectable in 92.3±1.0% (N=11) of PNA− ionocytes, and the abundance of PNA− ionocytes increased in response to systemic acidosis elicited by 72 h exposure to water of low pH (nominally pH 4.5), hypercapnia (1% CO2, nominally 7.6 Torr) or hyperoxia (achieved by gassing water with pure O2), as did the abundance of PNA− ionocytes that exhibited immunofluorescence for CAc. However, just 4.3 ± 0.6% (N=11) of PNA+ ionocytes expressed detectable CAc under control conditions. Marked increases in the abundance of CAc-positive PNA+ ionocytes were detected following exposure of trout to a base load via recovery from hypercapnia, or base infusion (72 h infusion with 140 mmol L−1 NaHCO3). The percentage of CAc-positive PNA+ ionocytes also was increased in trout treated with cortisol (10 mg kg−1 hydrocortisone 21-hemisuccinate daily for 7 d). These results suggest that regulation of CA within PNA+ ionocytes and/or the abundance of CAc-positive PNA+ ionocytes plays a role in activating base secretion in response to systemic alkalosis.