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Volume 66, Issue 2, Pages 633-640 (August 2004) Dopamine D2-like receptor agonist bromocriptine protects against ischemia/reperfusion injury in rat kidney Vihang Narkar, Oksana Kunduzova, Tahir Hussain, Claudie Cambon, Angelo Parini, Mustafa Lokhandwala Kidney International Volume 66, Issue 2, Pages 633-640 (August 2004) DOI: 10.1111/j.1523-1755.2004.00783.x Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 1 Schematic representation of induction of renal ischemic/reperfusion (I/R) injury. Kidney International 2004 66, 633-640DOI: (10.1111/j.1523-1755.2004.00783.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 2 Histology in cortical region of kidney slices of unilateral I/R rat model. Representative light micrographs (magnification, 40×) of hematoxylin and eosin stained sections showing morphologic features of cortical region 24 hours post I/R injury. In vehicle-treated rats, the ischemic cortical slices (B) showed extensive necrosis (indicated by arrows) compared to the contralateral kidney (A). The histomorphology of bromocriptine-treated ischemic kidneys (D) was normal and indistinguishable from contralateral control kidneys (C). Similar results were obtained from 5 animals. Kidney International 2004 66, 633-640DOI: (10.1111/j.1523-1755.2004.00783.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 3 Apoptosis in cortical region of kidney slices of unilateral ischemic/reperfusion (I/R) injury rat model. Apoptosis was determined in 24 hours post-I/R cortical kidney slices. Compared to the contralateral cortical slices (A), in vehicle-treated rats, the ischemic cortical slices (B) showed extensive TUNEL-positive staining. (C) The contralateral cortical slices of bromocriptine-treated rats are represented. In bromocriptine-treated rats low TUNEL-positive staining was detected in the ischemic cortical slices (D). (E) Data represented as % TUNEL-positive cells (N = 3). * and # represent statistically significant difference between groups compared (P < 0.05) [one-way analysis of variance (ANOVA); post hoc, Tukey's multiple comparision test]. Kidney International 2004 66, 633-640DOI: (10.1111/j.1523-1755.2004.00783.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 4 Detection of NKA activity and α1 subunits in renal cortical homogenates of unilateral ischemic/reperfusion (I/R) rat model. NKA activity and α1 subunit levels were determined in cortical homogenates obtained from the vehicle-treated and bromocriptine-treated rats 24 hours post-I/R. (A) In vehicle-treated rats, the NKA activity in ischemic cortical homogenate (IR) was significantly lower than in the contralateral cortical homogenate (Co). (B) In bromocriptine-treated rats, NKA activity was similar between contralateral and ischemic cortical homogenates. Data are represented as inorganic phosphate (Pi) released in μmol/mg/min. *Represents significant difference between Co and IR in vehicle-treated rats (P < 0.05) (unpaired Student t test). (C) Representative Western blot of NKA α1 subunit expression in cortical homogenates of vehicle-treated and bromocriptine-treated rats 24 hours post I/R. Similar results were obtained in N = 4 animals. Kidney International 2004 66, 633-640DOI: (10.1111/j.1523-1755.2004.00783.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 5 Determination of p44/42 mitogen-activated protein kinases (MAPKs) phosphorylation in renal cortical homogenates of unilateral ischemic/reperfusion (I/R) rat model. (A) In vehicle-treated rats, low phosphorylation was detected in contralateral (1) and ischemic (2) cortical homogenate. On the other hand, bromocriptine-treated rats showed extensive phosphorylation in contralateral (3) and ischemic (4) cortical homogenate (upper panel). The total-p42 MAPKs was similar in all the groups compared (1, 2, 3, and 4) (lower panel). The above are representative blots. Similar results were obtained from N = 4 sets of animals. (B) Data represented as average (mean ± SEM) ratio of phospho-p42 to total p42 MAPKs from N = 4 animals. (P < 0.05) [one-way analysis of variance (ANOVA); post hoc, Newman-Keuls multiple comparison test). Kidney International 2004 66, 633-640DOI: (10.1111/j.1523-1755.2004.00783.x) Copyright © 2004 International Society of Nephrology Terms and Conditions