AbstractVoltage‐sensitive calcium channels (VSCCs) are key regulators of osteoblast plasma membrane Ca2+ permeability and are under control of calcitropic hormones. Subtype specific antibodies were used to probe L‐type Cav1.2 (α1C) and T‐type Cav3.2 (α1H) subunit expression during mouse skeletal development. Commencing from E14.5 and continuing through skeletal maturity, immunoreactivity of Cav1.2 (α1C) subunits was evident in regions of rapid long bone growth, including the perichondrium, periosteum, chondro‐osseous junction and trabecular bones. Cav3.2 (α1H) subunits appeared simultaneously and followed a similar distribution pattern. Both subunits were observed in osteoblasts and chondrocytes under high magnification. Interestingly, Cav3.2 (α1H) subunits were present, but Cav1.2 (α1C) subunits were absent from osteocytes. Western Blot and immunohistochemical assessment of in vitro cell culture models of osteogenesis and chondrogenesis confirmed the in vivo observations. We conclude that both L‐type Cav1.2 (α1C) and T‐type Cav3.2 (α1H) VSCCs are dynamically regulated in bones and cartilages during endochondral bone development. Developmental Dynamics 234:54–62, 2005. © 2005 Wiley‐Liss, Inc.