Afferent Renal Nerve Activity is Proportionately Increased with Cystic Progression in a Preclinical Model of Autosomal Recessive Polycystic Kidney Disease
Authors: Gauthier Madeline , Dennis Melissa , Morales Mark , Banek Christopher T. 
Published: May 2022

Polycystic kidney disease (PKD) is the most common genetic cause of kidney failure, though the mechanisms underlying cyst formation and disease progression remain unclear. Innervation of the kidney contributes to the pathology of other cardiorenal dysfunction, including hypertension and chronic kidney disease ‐ frequent complications of PKD. In previous studies, we demonstrated renal denervation (RDNX), either total or afferent‐targeted (sensory) nerve ablation, reduced cystogenesis in a rat model of autosomal recessive PKD, thus highlighting a role for renal afferent nerves in PKD renal cystogenesis. To further elucidate this novel relationship, we aimed to quantify the renal afferent nerve activity (ARNA) in the adult PCK rat compared to non‐cystic controls. We hypothesized that ARNA would be elevated in the PCK rat compared to controls, and ARNA would directly correlate with renal cystic severity.

To address our hypothesis, we conducted direct afferent renal nerve recordings in 10‐week‐old PCK (n=15; 11M/4F) and age‐matched Sprague‐Dawley controls (SD; n=14; 6M/8F). Multiunit nerve recording was performed under isoflurane anesthesia by isolating and placing a renal nerve bundle on an iridium bipolar electrode, which was then encased in silicone. The nerve bundle was severed proximal to the electrode to isolate afferent renal nerve activity (ARNA). Following a 10‐minute stabilization period, resting ARNA was recorded for 10 minutes. To permit comparison between animals, resting ARNA was normalized to a maximal stimulus, achieved by renal pelvic perfusion of 50 µM capsaicin. Finally, electrical noise was isolated by sectioning the distal end of the nerve bundle. Recordings were rectified and integrated over 50 ms. ARNA was expressed as a percentage of maximal ARNA. Following nerve recording, kidneys were collected, formalin fixed, and embedded in paraffin for histologic analysis of renal cystic index (CI). Kidneys were stained with Masson’s Trichrome and analyzed using ImageJ, and CI was quantified by normalizing cystic area to the total kidney area. Data presented as mean ±SEM.

Resting ARNA was higher in PCK (22.7±4.2%) rats compared to non‐cystic SD controls (7.4±1.3%; Figure 1). Overall, PCK rats had an average CI of 8.7±0.7%. No sex differences in ARNA between male and female PCK rats were detected; however, CI was higher in male PCK rats (9.5±2.3%) compared to female PCK (6.3±1.9%). Further analysis of ARNA versus cystic index revealed a direct correlation between ARNA and renal cystic severity (F=5.9; R=0.61; p<0.05; Figure 2).

These findings support our hypothesis that ARNA is elevated in the PCK rat. Importantly, ARNA is directly correlated with cystic progression in this model. To our knowledge, this study is the first to directly measure ARNA in this model of ARPKD, as well as to compare ARNA to cyst severity. These findings, combined with our previous studies, lead us to conclude that increased ARNA may directly contribute to the progression of PKD in this model. Presently, these data are limited to single endpoint observations. We are currently conducting additional studies to further dissect the temporal relationship between ARNA and renal cyst development. Our future studies will investigate the stimuli amplifying ARNA in a continued effort to parse the role of this novel neural‐renal axis in the pathogenesis of PKD.