Protein Kinase C α Modulates the Ca²⁺ Influx Phase of the Ca²⁺ Response to 1α,25-Dihydroxy-Vitamin-D₃ in Skeletal Muscle Cells

Treatment of chick skeletal muscle cells with 1α,25-dihydroxy-vitamin D₃ [1α,25(OH)₂D₃] triggers a rapid and sustained increase in cytosolic Ca²⁺ ([Ca²⁺]_i), which depends on Ca²⁺ mobilization from inner stores and extracellular Ca²⁺ entry. Fluorimetric analysis of changes in [Ca²⁺]_i in Fura-2-loaded cells revealed that the hormone significantly stimulates the Ca²⁺ influx phase within the concentration range of 10^-12 - 10^-6 M, with maximal effects (3.5-fold increase) at 10^-9 M 1α,25(OH)₂D₃. The effects of the sterol on the Ca²⁺ entry pathway were abolished by the PKC inhibitors bisindolylmaleimide and calphostin. We have recently shown that, in these cells, 1α,25(OH)₂D₃ activates and translocates PKC α to the membrane, suggesting that this isozyme accounts for PKC-dependent 1α,25(OH)₂D₃ modulation of Ca²⁺ entry. The role of PKC α was specifically addressed here using antisense technology. When the expression of PKC α was selectively knocked out by intranuclear microinjection of an antisense oligonucleotide against PKC α mRNA, the Ca²⁺ influx component of the response to 1α,25(OH)₂D₃ was markedly reduced (- 60 %). These results demonstrate that 1α,25(OH)₂D₃-induced activation of PKC α enhances extracellular Ca²⁺ entry partially contributing to maintainance of the sustained phase of the Ca²⁺ response to the sterol.

Ca²⁺ Regulation (Skeletal Muscle) - 1α,25-Dihydroxy-Vitamin D₃ - PKC α Isozyme - Antisense Oligonucleotide

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