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Effects of adaptation to sea water, 170% sea water and to fresh water on activities and subcellular distribution of branchial Na+−K+-ATPase, low- and high affinity Ca++-ATPase, and ouabain-insensitive ATPase inGillichthys mirabilis

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Summary

  1. 1.

    Branchial activities of Na+−K+-ATPase, ouabain-insensitive ATPase, (Mg++-ATPase) and Ca++-ATPase were measured inGillichthys mirabilis after adaptation to salinities ranging from 170% SW to FW. Stabilities of these activities against freezing and deoxycholate solubilization and the temperature-dependence of activity rates were also investigated. Subcellular distribution and some kinetic properties of these activities, and of SDH were compared in branchial tissues of fish adapted to 170% SW and to FW.

  2. 2.

    Na+−K+-ATPase was maximally active at a Na+ concentration of 180 mM and K+ concentration of 60 mM. This enzyme was least active in 100% SW-adapted animals, but showed elevated activity after adaptation to 170% SW and to FW.Gillichthys is unusual (but not unique) among euryhaline teleosts by displaying higher Na+−K+-ATPase in FW- than in SW-adapted animals. Greatest activity, however, was observed in the heavy microsomal fraction (34,000xg) of the 170% SW-adapted group. Maximum SDH activity was also observed in the mitochondrial fraction (25,000xg) of 170% SW-adapted fish.

  3. 3.

    Activity of Ca++-ATPase displayed a complex Ca++-dependence. Two kinetic forms of this activity could be resolved, one with a high Ca++-affinity (K m=2.9 μM), the other having low Ca++-affinity (K m=0.88 mM). The low-affinity activity was reduced in branchial homogenates of 170% SW-compared with FW-adapted fish; the heavy microsomal fraction (34,000xg) derived from gills was most enriched in this Ca++-dependent ATPase in FW-adaptedGillichthys. In contrast, activities of the high-affinity form were equally enriched in heavy microsomal fractions of 170% SW- and FW-adapted animals. The high-affinity Ca++-ATPase was far more sensitive to inhibition by deoxycholate treatment than the low-affinity component. The possible roles of heterogeneous Ca++-dependent activities in branchial Ca++-transport are discussed.

  4. 4.

    Gill Mg++-ATPase activity was significantly elevated in FW and in 5% SW-adapted animals compared with the marine (170% SW- and 100% SW-adapted fish). The Mg++-dependent activity was most concentrated in the heavy microsomal fraction and to a lesser extent in the light microsomal and mitochondrial fractions in both 170% SW- and FW-adaptedGillichthys.

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Abbreviations

Ca++-ATPase:

calcium-activated adenosinetriphosphatase(EC 3.6.1.3.)

DMSO:

dimethylsulfoxide

EDTA:

ethylenediamine tetracetic acid

EGTA:

ethyleneglycol-bis- (aminoethyl ether) N,N′-tetracetic acid

ELON:

β-methylaminophenol sulfate

FW:

freshwater

HEPES:

N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid

Mg++-ATPase:

magnesium-activated adenosinetriphosphatase (EC 3.6.1.3)

Na+−K+-ATPase:

sodium, potassium-activated adenosinetriphosphatase (EC 3.6.1.3)

SDH:

succinate dehydrogenase (EC 1.3.99.1)

SW:

seawater

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  1. Division of Biological Sciences, University of Michigan, 48109, Ann Arbor, Michigan, USA

    Byron A. Doneen

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Supported by University of Michigan Phoenix Project and by NSF grant PCM-7922985

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Doneen, B.A. Effects of adaptation to sea water, 170% sea water and to fresh water on activities and subcellular distribution of branchial Na+−K+-ATPase, low- and high affinity Ca++-ATPase, and ouabain-insensitive ATPase inGillichthys mirabilis . J Comp Physiol B 145, 51–61 (1981). https://doi.org/10.1007/BF00782593

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