Laboratory Investigation

Kidney International (1995) 48, 1714–1720; doi:10.1038/ki.1995.469

Osmotic regulation of the betaine metabolism in immortalized renal cells

Rolf Willi Grunewald1 and Angela Eckstein1

1Abteilung Nephrologie, Universitätsklinik Göttingen, Göttingen, Germany

Correspondence: Dr R Willi Grunewald, Universitätsklinik Göttingen, Zentrum für Innere Medizin, Abt. für Nephrologie und Rheumatologie, Robert-Koch-Strabetae 40, D-37075 Göttingen, Germany.

Received 3 March 1994; Revised 23 June 1995; Accepted 25 June 1995.

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Abstract

Osmotic regulation of the betaine metabolism in immortalized renal cells. Betaine plays an important role in the osmoregulation of various renal cells. In the kidney betaine synthesis seems to be highest in the cortex, whereas osmotically regulated accumulation seems to play a crucial role in the inner medulla. Therefore, the influence of betaine synthesis on the long-term osmotic regulation of betaine content was investigated in epithelial SV40 transfected cell culture, derived from the outer medullary thick ascending limb of the loop of Henle (TALH) of rabbit kidney. Under hyperosmotic conditions the betaine content of TALH was significantly increased from 218 plusminus 35 micromol/g protein (300 mOsm/liter; control) to 334 plusminus 27 micromol/g (600 mOsm/liter; P < 0.0005). In addition the intracellular accumulation of 14C-betaine from 14C-choline was significantly elevated from 4.3 plusminus 1.0 micromol/g protein times hr) to 8.2 plusminus 1.0 micromol/g protein times hr; P 0.001) under hyperosmotic conditions. Synthesis of betaine was also influenced by the extracellular betaine content. In a betaine free medium the synthesis of betaine was increased by 7% (300 mOsm/liter; NS) or 40% (600 mOsm/liter; P < 0.0001) when compared to betaine containing medium. The alteration of betaine synthesis is presumably caused by osmotic regulation of the betaine aldehyde dehydrogenase. Activity of this enzyme was significantly higher under hyperosmotic conditions compared to isoosmotic control conditions (Vmax 4.1 plusminus 0.8 U/g protein; 600 mOsm/liter) versus 1.4 plusminus 0.1 U/g (300 mOsm/liter; P < 0.0001), while the affinity to betaine aldehyde remained unaltered. These results demonstrate that during long-term adaptation, betaine synthesis in TALH cells of the outer medulla of rabbit kidney can be regulated by extracellular osmolarity.

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