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Intracellular pH regulation by the plasma membrane V-ATPase in Malpighian tubules of Drosophila larvae

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Abstract

The functional significance of the apical vacuolar-type proton pump (V-ATPase) in Drosophila Malpighian tubules was studied by measuring the intracellular pH (pHi) and luminal pH (pHlu) with double-barrelled pH-microelectrodes in proximal segments of the larval anterior tubule immersed in nominally bicarbonate-free solutions (pHo 6.9). In proximal segments both pHi (7.43±0.20) and pHlu (7.10±0.24) were significantly lower than in distal segments (pHi 7.70±0.29, pHlu 8.09±0.15). Steady-state pHi of proximal segments was much less sensitive to changes in pHo than pH of the luminal fluid (ΔpHlu/ΔpHo was 0.49 while ΔpHi/ΔpHo was 0.18; pHo 6.50–7.20). Re-alkaliniziation from an NH4Cl-induced intracellular acid load (initial pHi recovery rate 0.55±0.34 pH·min-1) was nearly totally inhibited by 1 mmol·l-1 KCN (96% inhibition) and to a large degree (79%) by 1 μmol·l-1 bafilomycin A1. In contrast, both vanadate (1 mmol·l-1) and amiloride (1 mmol·l-1) inhibited pHi recovery by 38% and 33%, respectively. Unlike amiloride, removal of Na+ from the bathing saline had no effect on pHi recovery, indicating that a Na+/H+ exchange is not significantly involved in pHi regulation. Instead pHi regulation apparently depended largely on the availability of ATP and on the activity of the bafilomycin-sensitive proton pump.

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Abbreviations

DMSO:

dimethylsulphoxide

DNP:

2,4-dinitrophenol

NMDG:

N-methyl-D-glucamine

pHi :

intracellular pH

pHlu :

pH of the luminal fluid

pHo :

pH of the superfusion medium

βI :

intrinsic intracellular buffer capacity

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Authors and Affiliations

  1. Institut für Allgemeine und Spezielle Zoologie, Justus-Liebig-Universität, Stephanstrasse 24, D-35390, Giessen, Germany

    G. Bertram & A. Wessing

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  1. G. Bertram
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  2. A. Wessing
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Bertram, G., Wessing, A. Intracellular pH regulation by the plasma membrane V-ATPase in Malpighian tubules of Drosophila larvae. J Comp Physiol B 164, 238–246 (1994). https://doi.org/10.1007/BF00354085

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