Group VIA phospholipase A2 is a target for vasopressin signaling in the thick ascending limb.
Na(+)-K(+)-2Cl(-) cotransporter (NKCC2)-mediated NaCl reabsorption in the thick ascending limb (TAL) is stimulated by AVP via V2 receptor/PKA/cAMP signaling. This process is antagonized by locally produced eicosanoids such as 20-HETE or prostaglandin E(2), which are synthesized in a phospholipase A(2)-dependent reaction cascade. Using microarray-based gene expression analysis, we found evidence for an AVP-dependent downregulation of the calcium-independent isoform of PLA(2), iPLA(2)β, in the outer medulla of rats. In the present study, we therefore examined the contribution of iPLA(2)β to NKCC2 regulation. Immunoreactive iPLA(2)β protein was detected in cultured mTAL cells as well as in the entire TAL of rodents and humans with the exception of the macula densa. Administration of the V2 receptor-selective agonist desmopressin (5 ng/h; 3 days) to AVP-deficient diabetes insipidus rats increased outer medullary phosphorylated NKCC2 (pNKCC2) levels more than twofold in association with a marked reduction in iPLA(2)β abundance (-65%; P < 0.05), thus confirming microarray results. Inhibition of iPLA(2)β in Sprague-Dawley rats with FKGK 11 (0.5 μM) or in mTAL cells with FKGK 11 (10 μM) or (S)-bromoenol lactone (5 μM) for 1 h markedly increased pNKCC2 levels without affecting total NKCC2 expression. Collectively, these data indicate that iPLA(2)β acts as an inhibitory modulator of NKCC2 activity and suggest that downregulation of iPLA(2)β may be a relevant step in AVP-mediated urine concentration.
Dept. of Anatomy, Charité Universitätsmedizin-Berlin, Philippstrasse 12, Berlin, Germany. alexander.paliege@charite.de
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A
Paliege
T
T
Roeschel
H
H
Neymeyer
S
S
Seidel
T
T
Kahl
A L
AL
Daigeler
K
K
Mutig
R
R
Mrowka
N R
NR
Ferreri
B S
BS
Wilson
N
N
Himmerkus
M
M
Bleich
S
S
Bachmann
D000818:Animals; D000906:Antibodies; D001095:Arachidonic Acids; D002478:Cells, Cultured; D003894:Deamino Arginine Vasopressin; D015536:Down-Regulation; D005466:Fluorocarbons; D015870:Gene Expression; D054522:Group VI Phospholipases A2 / Q000276:immunology / Q000378:metabolism*; D006168:Guinea Pigs; D006801:Humans; D007527:Isoenzymes; D007659:Ketones; D007679:Kidney Medulla / Q000378:metabolism; D008138:Loop of Henle / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009281:Naphthalenes; D063065:Organophosphonates; D010766:Phosphorylation; D011753:Pyrones; D051381:Rats; D011910:Rats, Brattleboro; D017207:Rats, Sprague-Dawley; D028021:Sodium-Potassium-Chloride Symporters / Q000378:metabolism*; D064466:Solute Carrier Family 12, Member 1; D014667:Vasopressins / Q000378:metabolism*
C533806:1,1,1,2,2-pentafluoro-7-phenylheptan-3-one; D000906:Antibodies; D001095:Arachidonic Acids; D005466:Fluorocarbons; D007527:Isoenzymes; D007659:Ketones; D009281:Naphthalenes; D063065:Organophosphonates; D011753:Pyrones; C578714:SLC12A1 protein, human; C578715:Slc12a1 protein, mouse; C578716:Slc12a1 protein, rat; D028021:Sodium-Potassium-Chloride Symporters; D064466:Solute Carrier Family 12, Member 1; C104810:methyl arachidonylfluorophosphonate; D014667:Vasopressins; C068314:6-(bromomethylene)tetrahydro-3-(1-naphthaleneyl)-2H-pyran-2-one; D054522:Group VI Phospholipases A2; D003894:Deamino Arginine Vasopressin