Metabolic mode peculiar to Meckel’s cartilage: immunohistochemical comparisons of hypoxia‐inducible factor‐1α and glucose transporters in developing endochondral bones in mice

Abstract

The middle portion of Meckel's cartilage resembles endochondral bone formation accompanied by chondrocyte hypertrophy and death, cartilaginous matrix calcification, and chondroclastic resorption. We examined Meckel's cartilage specimens from mice mandibles taken on embryonic days 14-16 (E14-E16) using immunohistochemistry for hypoxia-inducible factor-1alpha (HIF-1alpha), glucose transporter 1 (GLUT1), glucose transporter 3 (GLUT3), and glucose transporter 5 (GLUT5), and using enzyme histochemistry for glucose-6-phosphate isomerase (GPI), lactate dehydrogenase (LDH), and cytochrome oxidase (COX), along with the periodic acid-Schiff (PAS) reaction, and compared the results with those of endochondral bones from E16 hind limbs. Periodic acid-Schiff-positive glycogen, HIF-1alpha, and GLUT immunoreactivity, and GPI, LDH, and COX activities were observed in Meckel's cartilage in E14 and E15 mandibles. In E16 mandibles, hypertrophic chondrocytes showed a transitory loss of HIF-1alpha immunoreactivity and consumed glycogen, while those closest to the resorption front showed intense immunoreactivity for HIF-1, GLUT3, and GLUT5. Hypertrophic chondrocytes of metatarsals possessed HIF-1alpha immunoreactivity in the nuclei and diminished COX activity, whereas developing tibias showed weak HIF-1alpha immunoreactivity even in hypoxic regions characterized by little or no COX activity. These findings suggest that HIF-1alpha becomes stabilized independently of the concentration of oxygen, and largely contributes to the development and resorption of Meckel's cartilage, probably through shifting the predominant metabolic mode from aerobic to anaerobic glycolysis.