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Cytoplasmic Ca2+-H+-exchange buffers in green algae

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Summary

Fluorescence ratio imaging was used for simultaneous measurement of cytosolic pHc and pCac inChara corallina, Nitella flexilis, andEremosphaera viridis. In some experiments the electrical membrane potential was also recorded. The first hint of coupling between changes in pHc and pCac was found in characean cells when the influence of butyrate on cytosolic streaming was studied by laser-Doppler-anemometry (LDA). The observed butyrate-induced cessation of cytosolic streaming supports the assumption that changes in pHc cause changes in pCac. This hypothesis was tested by simultaneously loading cells with Fura-2-dextran and BCECF-dextran. The addition of butyrate revealed strong coupling between pCac and pHc although this only occurred when the difference between pHc and pCac was less than 0.4 units (± 0.24, n=7). The measured relationship between the changes in pCac and pHc could be fitted by a cytoplasmic buffer exchange process. Protons imported by butyrate into the cytoplasm are able to displace Ca++ ions from cytoplasmic buffer sites. Three dissociation constants of the cytoplasmic buffer were pK 1=6.2, pK 2=7.1 for proton buffering, and pKca=6.7 for Ca++ ion buffering. Other possible mechanisms, such as butyrate-induced Ca++ influx through the plasmalemma and Ca++ release from internal stores are discussed. They are not necessary to explain the observed coupling but cannot be excluded from the process. Using the butyrate technique, the cytosolic in vivo proton buffer capacities ofN. flexilis, C. corallina, andE. viridis were determined as βi=30 mM · H+/pH-unit, βi=46 mM · H+/pH-unit, and βi=90 mM · H+/pH-unit, respectively. The values obtained in vivo are greater than those found previously using extraction methods. This can be explained in terms of pump activity and exchange with cell organelles, i.e., the vacuole. The high value of βi found inEremosphaera reflects adaptation to its habitat.

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Abbreviations

PPS:

protoplasmic streaming

HBA:

butyric acid

BCECF:

bis-carboxyethyl-carboxy-fluorescein

LDA:

laser-Doppler-anemometry

APW:

artificial pond water

CDPK:

calcium-dependent protein kinase

CICR:

calcium-induced calcium release

AP:

action potential

MES:

2-[N-morpholino]-ethane-sulfonic acid

PIPES:

piperazine-N,N′-bis[2-ethane-sulfonic acid]

EGTA:

ethylene-gly-col-tetra-acetic acid

HEPES:

N-[2-hydro-ethyl]-piperazine-N′-2-[ethane-sulfonic acid]

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  1. C. Plieth

    Present address: Department of Plant Sciences, University of Oxford, Oxford, UK

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  1. Institut für Angewandte Physik, Universität zu Kiel, Olshausenstrasse 40, D-24098, Kiel, Federal Republic of Germany

    C. Plieth & U. -P. Hansen

  2. Institutes for Plant Nutrition and Soil Sciences, University of Kiel, Kiel

    B. Sattelmacher

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Plieth, C., Sattelmacher, B. & Hansen, U.P. Cytoplasmic Ca2+-H+-exchange buffers in green algae. Protoplasma 198, 107–124 (1997). https://doi.org/10.1007/BF01282136

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