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This Article Full Text Full Text (PDF) All Versions of this Article: 293/3/F846    most recent 00139.2007v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Andratsch, M. Articles by Gstraunthaler, G. Search for Related Content PubMed PubMed Citation Articles by Andratsch, M. Articles by Gstraunthaler, G.

TGF- signaling and its effect on glutaminase expression in LLC-PK₁-FBPase⁺ cells

¹Department of Physiology and Medical Physics, Innsbruck Medical University, Innsbruck, Austria, and ²Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, Colorado

Submitted 27 March 2007 ; accepted in final form 21 June 2007

During systemic acidosis, renal proximal tubular cells exhibit enhanced rates of bicarbonate and ammonium ion synthesis and undergo extensive hypertrophy. The former adaptations are accomplished, in part, by increased expression of glutaminase (GA). LLC-PK₁-FBPase⁺ cells, a gluconeogenic line of porcine kidney cells, exhibit a rapid activation of the ERK1/2 and p38 MAPK pathways and a two- to threefold increase in GA mRNA when transferred to acidic medium (pH 6.9). Transforming growth factor- (TGF-), a potent activator of MAPK and Smad signaling cascades, also causes extensive renal hypertrophy. Thus the potential role of TGF- in the renal response to metabolic acidosis was investigated. Western blot analyses established that in LLC-PK₁-FBPase⁺ cells, TGF- activated the ERK1/2, p38 MAPK, and Smad1/5/8 pathways, but not the JNK and Smad2/3 pathways. Addition of TGF- to cells cultured in normal medium (pH 7.4) produced a steady increase in GA mRNA, resulting in a twofold induction after 18 h. Western blot analysis indicated that treatment with either TGF- or acidic medium resulted in an increased level of fibronectin. However, the effects of the two treatments on both GA mRNA and fibronectin levels occurred with different time courses and were additive. In addition, the rates of ammonia production were decreased slightly by addition of TGF-. Finally, a GA-luciferase reporter construct, which is activated 3.5-fold by treatment with acidic medium, is not affected by TGF-. Therefore, TGF- and metabolic acidosis activate some of the same signaling pathways in LLC-PK₁-FBPase⁺ cells, but produce separate effects on GA expression.

metabolic acidosis; renal hypertrophy; ERK1/2; p38 MAPK; Smad