Mechanism of Pdia3-dependent 1α,25-dihydroxy vitamin D3 signaling in musculoskeletal cells

1 alpha,25-Dihydroxy vitamin D3 [1,25(OH)2D3] acts on cells through traditional steroid hormone receptor-mediated gene transcription and by initiating rapid membrane-associated signaling pathways. Two receptors have been implicated in rapid signaling by 1,25(OH)2D3, the classical nuclear vitamin D receptor (VDR) and the more recently identified protein disulfide isomerase, family A, member 3 (Pdia3). Our lab along with other groups has established various tools to investigate the role of these two receptors, including gene knock-out, conditional knock-out, silencing, and over-expression in various model systems (growth plate chondrocytes, osteoblastic cells, chick intestinal epithelial cells, mouse embryoid bodies, extracellular matrix vesicles and isolated cell membranes). The data demonstrate the requirement for Pdia3 in 1,25(OH)2D3 induced phospholipase A2 (PLA2) and protein kinase C (PKC) activation and downstream responses. Pdia3+/- heterozygote mice also exhibit both cartilage and bone defects. VDR is present on the plasma membrane and one VDR-/- mouse strain lacks transcaltachia, although 1,25(OH)2D3 induced PKC activation and transcaltachia are not affected in another VDR-/- mouse strain. In the context of osteoblast differentiation, both receptors are expressed during osteogenic commitment of embryoid bodies and silencing of each causes a more mature osteoblast phenotype in MC3T3-E1 pre-osteoblasts. Pdia3 exists in caveolae, where it interacts with PLA2 activating protein (PLAA) and caveolin-1 to initiate rapid signaling via PLA2, phospholipase C (PLC), PKC, and ultimately the ERK1/2 family of mitogen activated protein kinases (MAPK). Using the growth plate chondrocyte and matrix vesicle models, we have demonstrated that Pdia3-dependent signaling in response to 1,25(OH)2D3 regulates growth plate physiology. (c) 2012 Elsevier Inc. All rights reserved.