Assessing intracellular pH regulation in H+-ATPase-rich ionocytes in zebrafish larvae using in vivo ratiometric imaging.

The Journal of Experimental Biology
H M YewSteve F Perry

Abstract

The H+-ATPase-rich (HR) cells of zebrafish larvae are a sub-type of ion-transporting cell located on the yolk sac epithelium that are responsible for Na+ uptake and H+ extrusion. Current models of HR cell ion transport mechanisms in zebrafish larvae are well established, but little is known about the involvement of the various ion transport pathways in regulating intracellular acid-base status. Here, a ratiometric imaging technique was developed and validated to monitor intracellular pH (pHi) continuously in larval zebrafish HR cells in vivo Gene knockdown or CRISPR/Cas9 knockout approaches were used to evaluate the roles of the two principal apical membrane acid excretory pathways, the Na+/H+ exchanger (NHE3b; slc9a3.2) and the H+-ATPase (atpv1aa). Additionally, the role of HR cell cytosolic carbonic anhydrase (CAc) was investigated because of its presumed role in providing H+ for Na+/H+ exchange and H+-ATPase. The temporal pattern and extent of intracellular acidification during exposure of fish to 1% CO2 and the extent of post-CO2 alkalisation were altered markedly in fish experiencing knockdown/knockout of CAc, NHE3b or H+-ATPase. Although there were slight differences among the three knockdown/knockout experiments, the typ...Continue Reading

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Citations

Oct 22, 2020·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Alex M ZimmerSteve F Perry
Mar 31, 2021·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Alex M ZimmerChris N Glover

Related Concepts

Metazoa
Proton-Translocating ATPase, F0 Sector
Hydrogen-Ion Concentration
Tadpole
Zebrafish
Antiporter Activity
Carbon Dioxide
Carbonate Dehydratase
Diagnostic Imaging
Proton-Translocating ATPases

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