Volume 58, Issue 5, Pages 1893-1901 (November 2000) Axial distribution and characterization of basolateral P2Y receptors along the rat renal tubule  Matthew A. Bailey, Martine Imbert-Teboul, Clare Turner, Sophie Marsy, Kaila Srai, Geoffrey Burnstock, Robert J. Unwin  Kidney International  Volume 58, Issue 5, Pages 1893-1901 (November 2000) DOI: 10.1111/j.1523-1755.2000.00361.x Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 1 (A) Sample trace showing the effect of removal of extracellular calcium on the response to adenosine 5′-triphosphate (ATP, 100 μmol/L) in isolated thin ascending limbs (ATL) and (B) peak ((?)) and plateau (□) responses [presented with basal [Ca2+]i subtracted (δ[Ca2+]i)] to superfusion of microdissected nephron segments with ATP (100 μmol/L). Data are means ± SE of 10 to 20 segments tested in 10 to 15 rats. Kidney International 2000 58, 1893-1901DOI: (10.1111/j.1523-1755.2000.00361.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 2 Representative traces showing the effect of ATP (100 μmol/L) and the phospholipase C-stimulating agonists noradrenaline (NAd; 10 μmol/L) and AVP (0.1 μmol/L) in the thin limbs of Henle. (A) In the descending thin limb (DTL), noradrenaline was typically less potent than ATP. (B) AVP and ATP were equipotent in the ascending thin limb (ATL). Kidney International 2000 58, 1893-1901DOI: (10.1111/j.1523-1755.2000.00361.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 3 (A) A representative trace of the ATP concentration-response curve in DTL and (B) ATP concentration-response curves for DTL (•), PCT (○), and OMCD (▵). (B) The data are presented with basal [Ca2+]i subtracted (δ[Ca2+]i) and are means ± SE of 5 to 15 segments tested in 5 to 10 rats. Kidney International 2000 58, 1893-1901DOI: (10.1111/j.1523-1755.2000.00361.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 4 The peak increase in cytosolic free calcium concentration elicited by 2MeSATP, ADP, ATP, ATPγS and UTP tested at 30 μmol/L. Up to three compounds only were used in any given segment. (A) In the PCT, 2MeSATP, ADP, and ATP were equipotent, whereas the effects of UTP and ATPγS were significantly lower (P < 0.05, analysis of variance), and (B) in the OMCD, all agents were equipotent (analysis of variance). Data shown in the histograms are presented with basal [Ca2+]i subtracted (δ[Ca2+]i) and are means ± SE of 5 to 19 segments tested in 5 to 12 rats. Kidney International 2000 58, 1893-1901DOI: (10.1111/j.1523-1755.2000.00361.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 5 The peak responses elicited by 2MeSATP, ADP, ATP, ATPγS, and UTP tested at 30 μmol/L in the (A) thin descending and (B) thin ascending limbs of Henle (DTL and ATL). Up to three agents were tested in the same segment. In both cases, ATP, UTP, and ATPγS were equipotent (analysis of variance), whereas 2MeSATP and ADP were without significant effect. These data are presented with basal [Ca2+]i subtracted (δ[Ca2+]i) and are means ± SE of four to eight segments tested in four to eight rats. Kidney International 2000 58, 1893-1901DOI: (10.1111/j.1523-1755.2000.00361.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 6 Sample gel from the RT-PCR experiments showing P2Y2 receptor expression in duplicate runs from each segment in the presence (RTase+) and absence (RTase-) of reverse transcriptase.[α-32P]dCTP was added to the PCR medium allowing relative quantitation of the PCR product in the different segments Table 4. Kidney International 2000 58, 1893-1901DOI: (10.1111/j.1523-1755.2000.00361.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 7 Sample gels from RT-PCR experiments showing the distribution of mRNA in isolated segments of the same rat. (A) P2Y1 receptor. (B) P2Y4 receptor. Similar data were obtained from three other rats. Kidney International 2000 58, 1893-1901DOI: (10.1111/j.1523-1755.2000.00361.x) Copyright © 2000 International Society of Nephrology Terms and Conditions