WNK-OSR1/SPAK-NCC signal cascade has circadian rhythm dependent on aldosterone

Oct 10, 2012Biochemical and biophysical research communications

Daily rhythm of the WNK-OSR1/SPAK-NCC signaling pathway depends on aldosterone

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Abstract

Phosphorylation levels of OSR1, SPAK, and NCC in mouse kidneys show diurnal rhythms.

The expression levels of WNK4, OSR1, SPAK, and NCC proteins did not display diurnal rhythms.
Phosphorylation of OSR1, SPAK, and NCC increased at the beginning of the active period and decreased at the start of the rest period.
Administration of eplerenone (10 mg/day) reduced the phosphorylation levels of these proteins.
Eplerenone also diminished the diurnal rhythm of NCC phosphorylation.
The diurnal rhythm in NCC phosphorylation may be influenced by aldosterone.

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Blood pressure and renal salt excretion show circadian rhythms. Recently, it has been clarified that clock genes regulate circadian rhythms of renal transporter expression in the kidney. Since we discovered the WNK-OSR1/SPAK-NaCl cotransporter (NCC) signal cascade, which is important for regulating salt balance and blood pressure, we have sought to determine whether NCC protein expression or phosphorylation shows diurnal rhythms in the mouse kidneys. Male C57BL/6J mice were sacrificed every 4h (at 20:00, 0:00, 4:00, 8:00, 12:00, and 16:00), and the expression and phosphorylation of WNK4, OSR1, SPAK, and NCC were determined by immunoblot. (Lights were turned on at 8:00, which was the start of the rest period, and turned off at 20:00, which was the start of the active period, since mice are nocturnal.) Although expression levels of each protein did not show diurnal rhythm, the phosphorylation levels of OSR1, SPAK, and NCC were increased around the start of the active period and decreased around the start of the rest period. Oral administration of eplerenone (10mg/day) attenuated the phosphorylation levels of these proteins and also diminished the diurnal rhythm of NCC phosphorylation. Thus, the activity of the WNK4-OSR1/SPAK-NCC cascade was shown to have a diurnal rhythm in the kidney that may be governed by aldosterone.

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WNK-OSR1/SPAK-NCC signal cascade has circadian rhythm dependent on aldosterone

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