Renal rescue of dopamine D2 receptor function reverses renal injury and high blood pressure
Prasad R. Konkalmatt, Laureano D. Asico, Yanrong Zhang, Yu Yang, Cinthia Drachenberg, Xiaoxu Zheng, Fei Han, Pedro A. Jose, Ines Armando
Prasad R. Konkalmatt, Laureano D. Asico, Yanrong Zhang, Yu Yang, Cinthia Drachenberg, Xiaoxu Zheng, Fei Han, Pedro A. Jose, Ines Armando
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Research Article Inflammation Nephrology

Renal rescue of dopamine D2 receptor function reverses renal injury and high blood pressure

Abstract

Dopamine D2 receptor (DRD2) deficiency increases renal inflammation and blood pressure in mice. We show here that long-term renal-selective silencing of Drd2 using siRNA increases renal expression of proinflammatory and profibrotic factors and blood pressure in mice. To determine the effects of renal-selective rescue of Drd2 expression in mice, the renal expression of DRD2 was first silenced using siRNA and 14 days later rescued by retrograde renal infusion of adeno-associated virus (AAV) vector with DRD2. Renal Drd2 siRNA treatment decreased the renal expression of DRD2 protein by 55%, and DRD2 AAV treatment increased the renal expression of DRD2 protein by 7.5- to 10-fold. Renal-selective DRD2 rescue reduced the expression of proinflammatory factors and kidney injury, preserved renal function, and normalized systolic and diastolic blood pressure. These results demonstrate that the deleterious effects of renal-selective Drd2 silencing on renal function and blood pressure were rescued by renal-selective overexpression of DRD2. Moreover, the deleterious effects of 45-minute bilateral ischemia/reperfusion on renal function and blood pressure in mice were ameliorated by a renal-selective increase in DRD2 expression by the retrograde ureteral infusion of DRD2 AAV immediately after the induction of ischemia/reperfusion injury. Thus, 14 days after ischemia/reperfusion injury, the renal expression of profibrotic factors, serum creatinine, and blood pressure were lower in mice infused with DRD2 AAV than in those infused with control AAV. These results indicate an important role of renal DRD2 in limiting renal injury and preserving normal renal function and blood pressure.

Authors

Prasad R. Konkalmatt, Laureano D. Asico, Yanrong Zhang, Yu Yang, Cinthia Drachenberg, Xiaoxu Zheng, Fei Han, Pedro A. Jose, Ines Armando

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Figure 1

Retrograde ureteral infusion of adeno-associated virus vectors provides renal-specific gene expression.

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Retrograde ureteral infusion of adeno-associated virus vectors provides ...
(A) Schematic representation of adeno-associated virus (AAV) vectors AAVLuc, AAVEGFP, and DRD2 AAV carrying the firefly luciferase cDNA, EGFP, or human dopamine receptor D2 (DRD2) cDNA driven by the CMV promoter. AAV-inverted terminal repeats (ITR) and SV40 polyadenylation signal (SV-pA) are also indicated. Mice were infused (B) systemically via the jugular vein or (C) retrogradely via the left ureter with ACMVLuc vector (1 × 1011 viral genome particles) carrying firefly luciferase under the control of CMV. Shown are representative bioluminescence images of the mouse in ventral and dorsal positions acquired on day 14 following vector administration. (D) Maximum and stable bioluminescence was achieved 14 days after injection. (E) Fourteen days following vector administration, the mice were euthanized and a panel of tissues (KR, right kidney; KL, left kidney; Sp, spleen; Lv, liver; Pa, pancreas; St, stomach; Sk, skeletal muscle; H, heart; Ln, lung; Br, brain) was collected. Luciferase activity in the indicated organs was determined using the in vitro luciferase assay kit (Promega Corporation) on protein extracts prepared as per the manufacturer’s guidelines. Luciferase activities are reported as RLU per mg protein. (F) AAV vector genome copy number in the indicated organs was determined by real-time quantitative PCR. n = 3/group.