Adenosine A₁receptor-operated calcium entry in renal afferent arterioles is dependent on postnatal maturation of TRPC3 channels

Abstract

Adenosine, a regulator of cardiovascular development and renal function, constricts renal afferent arterioles by inducing intracellular Ca concentration ([Ca]) elevation in smooth muscle cells (SMCs) via activation of its cognate A receptors (ARs). Mechanisms that underlie AR-dependent [Ca] elevation in renal vascular SMCs are not fully resolved. Whether AR expression and function in preglomerular microvessels are dependent on postnatal kidney maturation is also unclear. In this study, we show that selective activation of ARs by 2-chloro--cyclopentyladenosine (CCPA) does not stimulate store-operated Ca entry in afferent arterioles isolated from neonatal pigs. However, CCPA-induced [Ca] elevation is dependent on phospholipase C and transient receptor potential cation channel, subfamily C, member 3 (TRPC3). Basal [Ca] was unchanged in afferent arterioles isolated from newborn (0-day-old) pigs compared with their 20-day-old counterparts. By contrast, CCPA treatment resulted in significantly larger [Ca] in afferent arterioles from 20-day-old pigs. AR protein expression levels in the kidneys and afferent arterioles were unaltered in 0- vs. 20-day-old pigs. However, the TRPC3 channel protein expression level was ~92 and 78% higher in 20-day-old pig kidneys and afferent arterioles, respectively. These data suggest that activation of ARs elicits receptor-operated Ca entry in porcine afferent arterioles, the level of which is dependent on postnatal maturation of TRPC3 channels. We propose that TRPC3 channels may contribute to the physiology and pathophysiology of ARs.

γ‐secretase inhibitor DAPT mitigates cisplatin‐induced acute kidney injury by suppressing Notch1 signaling

“…Kidney tissue and HK‐2 cell samples were homogenized in ice‐cold RIPA buffer. Proteins were separated by SDS‐polyacrylamide gel (4%‐20%) electrophoresis as we have previously described . Immunoreactive proteins were visualized and documented using a gel documentation system (Bio‐Rad, Hercules, CA, USA).…”

Renal vascular TRP channels

“…SMC TRPC1 has been immunostained in rat afferent and efferent arterioles, although the study did not distinguish the level of TRPC1 expression between the arterioles ( Takenaka et al, 2002 ). TRPC3 channels are expressed in the plasma membrane of neonatal pig afferent arteriolar SMCs ( Soni et al, 2017a ). TRPC3 protein expression levels in whole kidneys and afferent arterioles were higher in 20-days-old than newborn pigs, suggesting postnatal changes in the expression of the channels ( Soni et al, 2017a ).…”

“…TRPC3 channels are expressed in the plasma membrane of neonatal pig afferent arteriolar SMCs ( Soni et al, 2017a ). TRPC3 protein expression levels in whole kidneys and afferent arterioles were higher in 20-days-old than newborn pigs, suggesting postnatal changes in the expression of the channels ( Soni et al, 2017a ). Whereas TRPC4 protein was detected in the rat descending vasa recta (DVR), TRPC5 was notably absent ( Lee-Kwon et al, 2005 ).…”

“…Selective adenosine A 1 -receptor (A 1 R) activator 2-chloro-N6 cyclopentyladenosine (CCPA) stimulated receptor-operated calcium entry (ROCE) in neonatal pig afferent arterioles via TRPC3 channels ( Soni et al, 2017a ). Evidence indicated that induction of ROCE by CCPA is dependent on kidney maturation, as shown by a more significant increase in intracellular Ca 2+ in 20 days-old piglets than in newborns, paralleling the increased protein expression of TRPC3 in the kidneys and afferent arterioles of the older pigs ( Soni et al, 2017a ). Postnatal kidney maturation did not alter A 1 R expression ( Soni et al, 2017a ).…”

Polymodal roles of TRPC3 channel in the kidney

“…TRPC3 is the most abundant TRPC channel in the pre-glomerular resistance vessels aka afferent arterioles, where it contributes to the Ca 2+ signaling and receptor-operated Ca 2+ entry in the vascular smooth muscle cells [ 58 ]. The magnitude of the adenosine receptor type 1 (A1R)-stimulated [Ca 2+ ] i elevations positively correlates with TRPC3 expression in afferent arterioles of postnatal and 20 days old piglets [ 59 ]. Moreover, activation of the calcium-sensing receptor (CaSR) by high extracellular Ca 2+ augmented proliferation of glomerular contractile mesangial cells via stimulation of TRPC3-mediated Ca 2+ influx [ 60 ].…”