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Role of PTH on Gene Expression in Osteocytes of Cortical Long Bone from Mature Mice

ABSTRACT: Femur and Tibia of Mice 3 months of age were dissected, growth plate removed, and serial digested with collagenase-trypsin to remove endosteal and periosteal cells. The cortical bone tubes, highly enriched in osteocye in their natural bone matrix environment where then cultured with and and with out PTH at 250nM for 24hr, RNA extracted and biotinylated cDNA prepared, and hybrized to Illumina MouseWG-6_V2 arrays. Data was analyzed using Genome Studio-Gene Expression Module from Illumina.

ORGANISM(S): Mus musculus

PROVIDER: GSE61146 | GEO | 2015/03/03

SECONDARY ACCESSION(S): PRJNA260350

REPOSITORIES: GEO

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Project description:The pathogenesis of primary hyperparathyroidism (I-HPT) and secondary hyperparathyroidism (II-PTH) remains to be elucidated. To characterize their pathophysiology, we investigated the effects of calcium and phosphate on cell proliferation and PTH release in an organ culture of parathyroid tissues. Dissected parathyroid tissues obtained from patients with I-HPT (adenoma) or II-PTH (nodular hyperplasia) were precultured on a collagen-coated membrane for 1-4 week. After exchanging the medium for one containing various concentrations of phosphate, PTH release and [3H]thymidine incorporation were studied. In contrast to dispersed parathyroid cells cultured in a monolayer, calcium decreased PTH release in a concentration-dependent manner in parathyroid tissues. Furthermore, when parathyroid tissues obtained from II-PTH were precultured for 1-4 weeks, PTH release and parathyroid cell proliferation were significantly increased in high-phosphate medium. These phosphate effects were also observed to a lesser extent in parathyroid tissues obtained from I-HPT, but there was no significant difference between I-HPT and II-HPT. Microarray analyses revealed that mRNA levels of PTH, CaSR, and VDR were well preserved, and several growth factors (e.g. TGF-beta1-induced protein) were abundantly expressed in II-PTH. Using organ cultures of hyperparathyroid tissues, in which PTH release and CaSR are well preserved for a prolonged period, we have demonstrated that phosphate stimulates parathyroid cell proliferation not only in II-PTH but also in I-HPT. Although the mechanism responsible for phosphate-induced cell proliferation remains to be elucidated, our in vitro findings suggest that both parathyroid tissues preserve to some extent a physiological response system to hyperphosphatemia as observed in normal parathyroid cells. These data will be published in Journal of Bone & Mineral Metabolism. Keywords: Organ culture experiments, dultured in low vs.high phosphate medium

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Role of PTH on Gene Expression in Osteocytes of Cortical Long Bone from Mature Mice

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