Pkd2 haploinsufficiency alters intracellular calcium regulation in vascular smooth muscle cells

Autosomal-dominant polycystic kidney disease is a multiorgan disease and its vascular manifestations are common and life-threatening. Despite this, little is known about their pathogenesis. Somatic mutations to the normal PKD allele in cystic epithelia and cyst development associated with the unstable PKD2(WS25) allele suggest a two-hit model of cystogenesis. However, it is unclear if this model can account for the cardiovascular pathology or if haploinsufficiency alone is disease-associated. In the present study, we found a decreased polycystin-2 (PC2, protein encoded by PKD2 gene) expression in PKD2( +/-) vessels, roughly half the wild-type level, and an enhanced level of intracranial vascular abnormalities in PKD2 (+/-) mice when induced to develop hypertension. Consistent with these observations, freshly dissociated PKD2 (+/-) vascular smooth muscle cells have significantly altered intracellular CA(2+) homeostasis. The resting [CA(2+)](i) is 17.1% lower in PKD2 (+/-) compared with wild-type cells (P=0.0003) and the total sarcoplasmic reticulum CA(2+) store (emptied by caffeine plus thapsigargin) is decreased (P<0.0001). The store operated CA(2+) (SOC) channel activity is also decreased in PKD2 (+/-) cells (P=0.008). These results indicate that inactivation of just one PKD2 allele is sufficient to significantly alter intracellular CA(2+) homeostasis, and that PC2 is necessary to maintain normal SOC activity and the SR CA(2+) store in VSMCs. Based on these findings, and the fact that [CA(2+)](i) signaling is essential to the regulation of contraction, production and secretion of extracellular matrix, cellular proliferation and Apoptosis, we propose that the abnormal intracellular CA(2+) regulation associated with PKD2 haploinsufficiency is directly related to the vascular phenotype.