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Direct Effects of 1,25-Dihydroxyvitamin D₃ and 24,25-Dihydroxyvitamin D3 on Growth Zone and Resting Zone Chondrocyte Membrane Alkaline Phosphatase and Phospholipase-A2 Specific Activities^*

ZVI SCHWARTZ, DIXIE L. SCHLADER, LARRY D. SWAIN and BARBARA D. BOYAN

Departments of Orthopedics (Z.S., D.L.S., L.D.S., B.D.B.), Periodontics (Z.S.), and Biochemistry (B.D.B.), University of Texas Health Science Center San Antonio, Texas 78284

Address all correspondence and requests for reprints to: Barbara D. Boyan, Ph.D., University Industry Cooperative Research Center, University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, Texas 78284.

Abstract

1,25-Dihydroxyvitamin D₃ [1,25-(OH)₂D₃] and 24,25-(OH)₂D₃ differentially affect the specific activity of alkaline phosphatase (ALPase) and phospholipase-A₂ (PLA₂) of plasma membranes and extracellular matrix vesicles produced by costochondral reserve zone and growth zone cartilage chondrocytes in culture. In the present study, growth zone and cartilage and reserve zone matrix vesicles and plasma membranes were isolated from confluent chondrocyte cultures and incubated with hormone for 3 and 24 h in vitro. Addition of 1,25-(OH)₂D₃ to GC matrix vesicles and plasma membranes resulted in dose-dependent increases in ALPase and PLA₂ specific activities in both membrane fractions. Addition of 24,25- (OH)₂D₃ to RC membrane fractions stimulated matrix vesicle ALPase at 10^-7 and 10^-8 M and plasma membrane ALPase at 10^-8 M only. However, 24,25-(OH)₂D₃ inhibited matrix vesicle and plasma membrane PLA₂ activity. The effects of the vitamin D metabolites were noticed after both 3 and 24 h. Neither hormone metabolite had any effect on these enzymes in membrane fractions from cultures of neonatal rat muscle mesenchymal cells, which do not calcify their matrix in vivo. These data suggest that 1,25-(OH)₂D₃ and 24,25-(OH)₂D₃ can directly affect chondrocyte membrane enzymes without genomic influence or protein synthesis and that membrane response depends on the stage of chondrocyte differentiation. Changes in PLA₂ activity may change membrane fluidity and may be a mechanism by which the hormones affect cell membranes. (Endocrinology 123: 2878–2884 1988)

Footnotes

^* This work was supported by USPHS Grants DE-05932 and DE-05937.

NIH Minority Biomedical Research Support fellow at Incarnate Word College.

Received June 10, 1988.

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