Retinoic acid induces a shift in the energetic state of hypertrophic chondrocytes.
In the epiphyseal growth plate, chondrocyte maturation is accompanied by dramatic alterations in energy metabolism. To explore the relationship between these two events, we used retinoic acid (RA) to promote chondrocyte maturation in culture. The specific question that was addressed was, does RA treatment of cultured chondrocytes in vitro induce a change in energy status similar to that seen in hypertrophic chondrocytes in vivo. Maturing chondrocytes isolated from the cephalic region of day 18 chick embryo sterna were allowed to grow for 7-14 days in monolayer until confluent and then treated with 10-300 nM RA. Immature chondrocytes from the caudal region of sternum were grown in parallel and served as control cells for the study. We found that in maturing cephalic cell cultures, RA had a rapid and profound effect on oxidative metabolism. The retinoid caused a reduction in the energy charge ratio (ECR) and the ATP/ADP ratio and a sharp decrease in cell ATP levels. Maximum inhibition was observed when the RA concentration was 10-35 nM. Compared with the adenine nucleotides, creatine phosphate levels were decreased to a lesser extent by RA, although there was substantial inhibition of creatine kinase activity. We expected to find a compensatory elevation in glycolytic activities; however, the lactate levels in the medium of the treated cells indicated that anaerobic glycolysis was depressed. In contrast to the cephalic chondrocytes, when caudal cell cultures were treated with RA, lactate formation was stimulated and there were minimal effects on oxidative metabolism. To determine the mechanism of inhibition of glycolysis, we measured the activity of pyruvate kinase in RA-treated cephalic cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia.
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Shapiro
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Debolt
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Hatori
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Iwamoto
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Pacifici
D000255:Adenosine Triphosphate / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D004734:Energy Metabolism / Q000187:drug effects*; D006019:Glycolysis / Q000187:drug effects; D006132:Growth Plate / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D010725:Phosphocreatine / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors; D014212:Tretinoin / Q000494:pharmacology*
D010725:Phosphocreatine; D014212:Tretinoin; D000255:Adenosine Triphosphate; D000469:Alkaline Phosphatase