Blood and gill responses to HCl infusions in the Pacific hagfish (Eptatretus stoutii)

Abstract

Pacific hagfish ( Eptatretus stoutii (Lockington, 1878)) were serially infused with HCl (6000 µmol·kg^–1) every 6 h over a 24 h period to explore branchial acid secretion mechanisms. Blood pH recovered from a value ~2 pH units below that of resting pH. This pH regulatory mechanism was activated by 6 h and persisted over the 24 h time course. Western-blot analysis revealed an increased abundance (~2.3×) of NHE2-like immunoreactivity (NHE2 L-IR) in whole gill fractions from HCl-infused fish compared with control NaCl-infused fish. Membrane fractions isolated from the gills of HCl-infused fish also showed an increased abundance (~1.8×). Immunohistochemistry results demonstrated a significant redistribution of NHE2 L-IR to the apical region in HCl-infused fish. V-H⁺-ATPase abundance did not significantly increase in HCl-infused fish, while levels of Na⁺/K⁺-ATPase expression were unchanged. Immunohistochemistry of these transporters did not show any changes in cellular localization. We propose that the Pacific hagfish overcomes a metabolic acidosis via an increased synthesis of an NHE-like protein and an increased density of the transporter at the apical region of the mitochondria-rich cells of the gill.