Intracellular pH regulation in mantle epithelial cells of the Pacific oyster, Crassostrea gigas

J Comp Physiol B. 2020 Nov;190(6):691-700. doi: 10.1007/s00360-020-01303-3. Epub 2020 Aug 20.

Abstract

Shell formation and repair occurs under the control of mantle epithelial cells in bivalve molluscs. However, limited information is available on the precise acid-base regulatory machinery present within these cells, which are fundamental to calcification. Here, we isolate mantle epithelial cells from the Pacific oyster, Crassostrea gigas and utilise live cell imaging in combination with the fluorescent dye, BCECF-AM to study intracellular pH (pHi) regulation. To elucidate the involvement of various ion transport mechanisms, modified seawater solutions (low sodium, low bicarbonate) and specific inhibitors for acid-base proteins were used. Diminished pH recovery in the absence of Na+ and under inhibition of sodium/hydrogen exchangers (NHEs) implicate the involvement of a sodium dependent cellular proton extrusion mechanism. In addition, pH recovery was reduced under inhibition of carbonic anhydrases. These data provide the foundation for a better understanding of acid-base regulation underlying the physiology of calcification in bivalves.

Keywords: Acid–base; Calcification; Ion transport proteins; Mollusc.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetazolamide / pharmacology
  • Amiloride / analogs & derivatives
  • Amiloride / pharmacology
  • Animals
  • Calcification, Physiologic
  • Carbonic Anhydrase Inhibitors / pharmacology
  • Crassostrea*
  • Cytophotometry
  • Epithelial Cells / chemistry*
  • Epithelial Cells / drug effects
  • Hydrogen-Ion Concentration
  • Ion Transport
  • Sodium Channel Blockers / pharmacology
  • Sodium-Hydrogen Exchangers / antagonists & inhibitors

Substances

  • Carbonic Anhydrase Inhibitors
  • Sodium Channel Blockers
  • Sodium-Hydrogen Exchangers
  • Amiloride
  • Acetazolamide
  • ethylisopropylamiloride