Skip to Content
MilliporeSigma
  • Altered signaling pathways linked to angiotensin II underpin the upregulation of renal Na(+)-ATPase in chronically undernourished rats.

Altered signaling pathways linked to angiotensin II underpin the upregulation of renal Na(+)-ATPase in chronically undernourished rats.

Biochimica et biophysica acta (2014-10-07)
Paulo A Silva, Humberto Muzi-Filho, Amaury Pereira-Acácio, Juliana Dias, João F S Martins, Maicon Landim-Vieira, Karine S Verdoorn, Lucienne S Lara, Leucio D Vieira-Filho, Edjair V Cabral, Ana D O Paixão, Adalberto Vieyra
ABSTRACT

This study has investigated the participation of altered signaling linked to angiotensin II (Ang II) that could be associated with increased Na(+) reabsorption in renal proximal tubules during chronic undernutrition. A multideficient chow for rats (basic regional diet, BRD) was used, which mimics several human diets widely taken in developing countries. The Vmax of the ouabain-resistant Na(+)-ATPase resident in the basolateral membranes increased >3-fold (P<0.001) accompanied by an increase in Na(+) affinity from 4.0 to 0.2mM (P<0.001). BRD rats had a >3-fold acceleration of the formation of phosphorylated intermediates in the early stage of the catalytic cycle (in the E1 conformation) (P<0.001). Immunostaining showed a huge increase in Ang II-positive cells in the cortical tubulointerstitium neighboring the basolateral membranes (>6-fold, P<0.001). PKC isoforms (α, ε, λ, ζ), Ang II type 1 receptors and PP2A were upregulated in BRD rats (in %): 55 (P<0.001); 35 (P<0.01); 125, 55, 11 and 30 (P<0.001). PKA was downregulated by 55% (P<0.001). With NetPhosK 1.0 and NetPhos 2.0, we detected 4 high-score (>0.70) regulatory phosphorylation sites for PKC and 1 for PKA in the primary sequence of the Na(+)-ATPase α-subunit, which are located in domains that are key for Na(+) binding and catalysis. Therefore, chronic undernutrition stimulates tubulointerstitial activity of Ang II and impairs PKC- and PKA-mediated regulatory phosphorylation, which culminates in an exaggerated Na(+) reabsorption across the proximal tubular epithelium.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-PP2A, C subunit antibody, Mouse monoclonal, clone 7A6, purified from hybridoma cell culture
Sigma-Aldrich
Methyl Green, zinc chloride salt, for microscopy (Bact., Bot., Hist.)
Sigma-Aldrich
Methyl Green, zinc chloride salt, <0.5% crystal violet, Dye content 80 %
Sigma-Aldrich
Methyl Green, zinc chloride salt, certified by the Biological Stain Commission, Dye content 85 %
Sigma-Aldrich
Methyl Green, zinc chloride salt, ~85%
Supelco
o-Xylene, analytical standard
Sigma-Aldrich
o-Xylene, anhydrous, 97%
Supelco
o-Xylene, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
o-Xylene, reagent grade, ≥98.0%
Sigma-Aldrich
o-Xylene, puriss. p.a., ≥99.0% (GC)
Supelco
o-Xylene, suitable for HPLC, 98%