Kinetics and osmoregulation of Na +- And Cl -dependent betaine transporter in rat renal medulla

Gilbert W. Moeckel, Li-Wen Lai, Walter G. Guder, H. Moo Kwon, Yeong Hau H Lien

Research output: Contribution to journalArticle

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Abstract

-Betaine is one of the maior organic osmolytes that accumulate in the renal medulla in response to high extracellular tonicity. Recent studies in MDCK cells have shown that betaine is taken up by an Na +- and Cl~-dependent transporter located on the basolateral membrane. We demonstrate here the presence of Na +-Cl"dependent betaine transporter(s) in tubule suspensions prepared from the rat outer and inner medulla. The betaine transport activity was two to three times higher in the inner medulla compared with the outer medulla. The removal of Na + and Cl~ reduced betaine uptake in the outer medullary tubules by 86% and 82%, respectively. The betaine uptake was decreased by 39% in hypotonie buffer (189 mosmol/ kgH2O) and increased by 82% in hypertonic buffer (545 mosmol/kgHO), compared with isotonic buffer (308 mosmol/ kgH2O). Kinetic studies of Na +-dependent betaine uptake in the outer medullary tubules revealed both a low- and a high-affinity component as follows: low-affinity and highvolume component with Michaelis constant (Kmi) of 8.6 mM and maximal uptake rate (Vmaxi) of 112 pmol -ug protein"1 -h"1; and a low-volume and high-affinity component with Km% of 0.141 mM and Vmax2 of 10 pmol-pg protein"'-h"1. To investigate whether the Na +-Cl"-dependent betaine transporter is regulated by tonicity in vivo, we quantitated its mRNA in rat renal cortex and outer and inner medulla using both canine and rat Na +-Cl~-dependent betaine transporter cDNAprobes. A single band of 3.0 kb was seen in the Northern blots prepared from both outer and inner medulla, but not in the cortex. Water deprivation for 3 days increased the abundance of this mRNA in the outer and inner medulla by 140% and 170%, respectively, but did not affect its expression in the cortex. In conclusion, Na +-Cl~-dependent betaine transporter(s) is present in rat outer and inner medullary tubules, and betaine transporter mRNA abundance is regulated by the hydration state in vivo.

Original languageEnglish (US)
JournalThe American journal of physiology
Volume272
Issue number1 PART 2
StatePublished - 1997

Fingerprint

Osmoregulation
Betaine
Kidney
Buffers
Messenger RNA
Water Deprivation
Madin Darby Canine Kidney Cells
Northern Blotting
betaine plasma membrane transport proteins
Canidae
Suspensions
Proteins
Membranes

Keywords

  • Betaine
  • Betaine transport
  • Dehydration
  • Gene regulation
  • Osmolytes
  • Y-aminobutyric acid transporter

ASJC Scopus subject areas

  • Physiology (medical)

Cite this

Kinetics and osmoregulation of Na +- And Cl -dependent betaine transporter in rat renal medulla. / Moeckel, Gilbert W.; Lai, Li-Wen; Guder, Walter G.; Moo Kwon, H.; Lien, Yeong Hau H.

In: The American journal of physiology, Vol. 272, No. 1 PART 2, 1997.

Research output: Contribution to journalArticle

Moeckel, Gilbert W. ; Lai, Li-Wen ; Guder, Walter G. ; Moo Kwon, H. ; Lien, Yeong Hau H. / Kinetics and osmoregulation of Na +- And Cl -dependent betaine transporter in rat renal medulla. In: The American journal of physiology. 1997 ; Vol. 272, No. 1 PART 2.
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T1 - Kinetics and osmoregulation of Na +- And Cl -dependent betaine transporter in rat renal medulla

AU - Moeckel, Gilbert W.

AU - Lai, Li-Wen

AU - Guder, Walter G.

AU - Moo Kwon, H.

AU - Lien, Yeong Hau H

PY - 1997

Y1 - 1997

N2 - -Betaine is one of the maior organic osmolytes that accumulate in the renal medulla in response to high extracellular tonicity. Recent studies in MDCK cells have shown that betaine is taken up by an Na +- and Cl~-dependent transporter located on the basolateral membrane. We demonstrate here the presence of Na +-Cl"dependent betaine transporter(s) in tubule suspensions prepared from the rat outer and inner medulla. The betaine transport activity was two to three times higher in the inner medulla compared with the outer medulla. The removal of Na + and Cl~ reduced betaine uptake in the outer medullary tubules by 86% and 82%, respectively. The betaine uptake was decreased by 39% in hypotonie buffer (189 mosmol/ kgH2O) and increased by 82% in hypertonic buffer (545 mosmol/kgHO), compared with isotonic buffer (308 mosmol/ kgH2O). Kinetic studies of Na +-dependent betaine uptake in the outer medullary tubules revealed both a low- and a high-affinity component as follows: low-affinity and highvolume component with Michaelis constant (Kmi) of 8.6 mM and maximal uptake rate (Vmaxi) of 112 pmol -ug protein"1 -h"1; and a low-volume and high-affinity component with Km% of 0.141 mM and Vmax2 of 10 pmol-pg protein"'-h"1. To investigate whether the Na +-Cl"-dependent betaine transporter is regulated by tonicity in vivo, we quantitated its mRNA in rat renal cortex and outer and inner medulla using both canine and rat Na +-Cl~-dependent betaine transporter cDNAprobes. A single band of 3.0 kb was seen in the Northern blots prepared from both outer and inner medulla, but not in the cortex. Water deprivation for 3 days increased the abundance of this mRNA in the outer and inner medulla by 140% and 170%, respectively, but did not affect its expression in the cortex. In conclusion, Na +-Cl~-dependent betaine transporter(s) is present in rat outer and inner medullary tubules, and betaine transporter mRNA abundance is regulated by the hydration state in vivo.

AB - -Betaine is one of the maior organic osmolytes that accumulate in the renal medulla in response to high extracellular tonicity. Recent studies in MDCK cells have shown that betaine is taken up by an Na +- and Cl~-dependent transporter located on the basolateral membrane. We demonstrate here the presence of Na +-Cl"dependent betaine transporter(s) in tubule suspensions prepared from the rat outer and inner medulla. The betaine transport activity was two to three times higher in the inner medulla compared with the outer medulla. The removal of Na + and Cl~ reduced betaine uptake in the outer medullary tubules by 86% and 82%, respectively. The betaine uptake was decreased by 39% in hypotonie buffer (189 mosmol/ kgH2O) and increased by 82% in hypertonic buffer (545 mosmol/kgHO), compared with isotonic buffer (308 mosmol/ kgH2O). Kinetic studies of Na +-dependent betaine uptake in the outer medullary tubules revealed both a low- and a high-affinity component as follows: low-affinity and highvolume component with Michaelis constant (Kmi) of 8.6 mM and maximal uptake rate (Vmaxi) of 112 pmol -ug protein"1 -h"1; and a low-volume and high-affinity component with Km% of 0.141 mM and Vmax2 of 10 pmol-pg protein"'-h"1. To investigate whether the Na +-Cl"-dependent betaine transporter is regulated by tonicity in vivo, we quantitated its mRNA in rat renal cortex and outer and inner medulla using both canine and rat Na +-Cl~-dependent betaine transporter cDNAprobes. A single band of 3.0 kb was seen in the Northern blots prepared from both outer and inner medulla, but not in the cortex. Water deprivation for 3 days increased the abundance of this mRNA in the outer and inner medulla by 140% and 170%, respectively, but did not affect its expression in the cortex. In conclusion, Na +-Cl~-dependent betaine transporter(s) is present in rat outer and inner medullary tubules, and betaine transporter mRNA abundance is regulated by the hydration state in vivo.

KW - Betaine

KW - Betaine transport

KW - Dehydration

KW - Gene regulation

KW - Osmolytes

KW - Y-aminobutyric acid transporter

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