Janet D. Klein, Robert B. Gunn, Brian R. Roberts, Jeff M. Sands

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Down-regulation of urea transporters in the renal inner medulla of lithium-fed rats Janet D. Klein, Robert B. Gunn, Brian R. Roberts, Jeff M. Sands Kidney International Volume 61, Issue 3, Pages 995-1002 (March 2002) DOI: 10.1046/j.1523-1755.2002.00210.x Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 1 Aquaporin-2 (AQP2) protein abundance in kidney inner medullary (IM) tip (A) and base (B) from control rats and rats fed LiCl for 25 days. Bottom panels are representative Western blots showing AQP2 protein bands in IM tip (A) and base (B) from control (C) and LiCl-fed (Li) rats. Each lane represents a sample from a separate rat. Top panels are a densitometric summary of the sum of the 35-45 kD (glycosylated) and 29 kD (non-glycosylated) AQP2 bands on the gels. AQP2 protein abundance was significantly reduced in LiCl-fed rats in both the IM tip and base, compared to control rats. Data are mean ± SE, N = 4 rats/group; *P < 0.05. Kidney International 2002 61, 995-1002DOI: (10.1046/j.1523-1755.2002.00210.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 2 UT-A1 urea transporter protein abundance in kidney inner medullary (IM) tip from control rats and rats fed LiCl for 25 days. (A) Representative Western blot showing UT-A1 protein bands in IM tip from control (C) and LiCl-fed (Li) rats. Each lane represents a sample from a separate rat. (B) Densitometric summary of the sum of the 117 and 97 kD bands on the gels. There was a significant decrease in the density of the 117 and 97 kD bands in LiCl-fed rats compared to control rats. Data are mean ± SE, N = 8 rats/group; *P < 0.05. Kidney International 2002 61, 995-1002DOI: (10.1046/j.1523-1755.2002.00210.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 3 UT-A1 urea transporter protein abundance in kidney inner medullary (IM) base from control rats and rats fed LiCl for 25 days. (A) Representative Western blot showing UT-A1 protein bands in IM base from control (C) and LiCl-fed (Li) rats. Each lane represents a sample from a separate rat. (B) Densitometric summary of the 97 kD band on the gels. There was a significant decrease in the density of the 97 kD band in LiCl-fed rats compared to control rats. Data are mean ± SE, N = 8 rats/group; *P < 0.05. Kidney International 2002 61, 995-1002DOI: (10.1046/j.1523-1755.2002.00210.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 4 UT-B urea transporter protein abundance in kidney inner medullary (IM) base from control rats and rats fed LiCl for 25 days. (A) Representative Western blot showing UT-B protein bands in IM base from control (C) and LiCl-fed (Li) rats. Each lane represents a sample from a separate rat. (B) Densitometric summary of the 40 to 50 kD band on the gels. There was a significant decrease in the density of the 40 to 50 kD band in LiCl-fed rats compared to control rats. Data are mean ± SE, N = 6 rats/group; *P < 0.05. Kidney International 2002 61, 995-1002DOI: (10.1046/j.1523-1755.2002.00210.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 5 Phosphorylation of UT-A1 in inner medullary collecting duct (IMCD) suspensions from control rats (A) and rats fed LiCl for 10 days (B) in response to 100 pmol/L vasopressin. A representative autoradiogram is presented showing that 100 pmol/L vasopressin increases 32PO4-incorporation into the 117 and 97 kD UT-A1 proteins in IMCDs from control rats but not from rats fed lithium. This result was repeated in two additional pairs of lithium-fed rats (data not shown). Kidney International 2002 61, 995-1002DOI: (10.1046/j.1523-1755.2002.00210.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 6 Phosphorylation of UT-A1 in IMCD suspensions from control rats (B) and rats fed LiCl for 10 days (A) in response to 100 nmol/L vasopressin. The top panels show representative autoradiograms showing that 100 nmol/L vasopressin increases 32PO4-incorporation into the 117 and 97 kD UT-A1 proteins in IMCDs from rats fed lithium, similar to the response in IMCDs from control rats. Each lane represents a sample from a separate rat. Bottom panel A is a densitometric summary of the 117 and 97 kD bands on the autoradiograms. There was a significant increase in the 32PO4-incorporation of the 117 and 97 kD band in LiCl-fed rats. Data are mean ± SE, N = 3 lithium-fed rats/group; *P < 0.05. Kidney International 2002 61, 995-1002DOI: (10.1046/j.1523-1755.2002.00210.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 7 UT-A urea transporter protein abundance in liver from control rats and rats fed LiCl for 10 days. (A) Representative Western blot showing UT-A protein bands in liver from control (C) and LiCl-fed (Li) rats. Each lane represents a sample from a separate rat. (B) Densitometric summary of the 49 kD band on the gels. There was a significant increase in the density of the 49 kD band in LiCl-fed rats compared to control rats. Data are mean ± SE, N = 6 rats/group; *P < 0.05. Kidney International 2002 61, 995-1002DOI: (10.1046/j.1523-1755.2002.00210.x) Copyright © 2002 International Society of Nephrology Terms and Conditions