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J. Biol. Chem., Vol. 268, Issue 6, 3821-3824, 02, 1993
S Uchida, S Sasaki, T Furukawa, M Hiraoka, T Imai, Y Hirata and F Marumo
Complementary DNA encoding a rat kidney chloride channel (CIC-K1) was
isolated by a polymerase chain reaction (PCR) cloning strategy. We designed
degenerate primers, based on the regions where previously cloned chloride
channels (CIC-0, -1, and -2) possess significant amino acid identity, and
performed reverse transcription PCR with whole kidney mRNA. The 686-amino
acid protein encoded by CIC-K1 is about 40% identical to the previously
cloned chloride channels and has a similar hydropathy profile. Expression
of CIC-K1 in Xenopus oocytes induced Cl- currents that activate
instantaneously upon hyperpolarization and depolarization, and displayed a
slightly outwardly rectifying current- voltage relationship. The message
for CIC-K1 was 2.4 kilobases and was found predominantly in kidney,
especially in the inner medulla. Reverse transcription PCR technique using
micro-dissected nephron segments revealed that the main site of expression
in kidney was the thin ascending limb of Henle's loop, which has the
highest Cl- permeability among the nephron segments and is thought to be
involved in a counter- current system for urine concentration in the inner
medulla. The abundance of CIC-K1 mRNA in kidney increased about 4-fold as
rats became dehydrated by deprivation of water for 5 days. The site of
expression and the regulation by dehydration suggest that CIC-K1 function
may be important in urinary concentrating mechanisms.
Molecular cloning of a chloride channel that is regulated by dehydration and expressed predominantly in kidney medulla [published erratum appears in J Biol Chem 1994 Jul 22;269(29):19192]
Second Department of Internal Medicine, Tokyo Medical and Dental University, Japan.
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