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J. Biol. Chem., Vol. 275, Issue 44, 34252-34259, November 3, 2000

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Expression of Functional Metabotropic Glutamate Receptors in Primary Cultured Rat Osteoblasts
CROSS-TALK WITH N-METHYL-D-ASPARTATE RECEPTORS^*

Yuchun Gu and Stephen J. Publicover^§

From the School of Biosciences, University of Birmingham, Birmingham B15 2TT, United Kingdom

Osteoblasts and osteoclasts express functional N-methyl-D-aspartate (NMDA) receptors, which participate in regulation of bone matrix. In rat femoral osteoblasts held in whole cell clamp there is a robust NMDA current but little if any response to L-glutamate. We have investigated expression of metabotropic glutamate receptors (mGluRs) in these cells. By reverse transcription polymerase chain reaction, we have detected expression of mGluR1b (but not mGluR1a, 2, 3, 4, 5, or 6). Blockade of mGluRs with (±)--methyl-carboxyphenyl-glycine resulted in an enlarged L-glutamate-induced current that resembled the response to NMDA. Conversely, prior stimulation of mGluRs with trans-(±)-1-amino-1,3-cyclopentanedicarboxylic acid (1S,3R-ACPD; mGluR agonist) reduced the NMDA-induced current by 77%. Monitoring of [Ca²⁺]_i showed that NMDA induced a sustained elevation of [Ca²⁺]_i, which was dependent upon [Ca²⁺]_o. Treatment with 1S,3R-ACPD generated an initial transient that was independent of [Ca²⁺]_o, followed by a sustained, [Ca²⁺]_o-dependent phase, a response consistent with phospholipase C-mediated mobilization of stored Ca²⁺. Investigations of the interaction between the two receptors confirmed inhibitory modulation of the NMDA receptor-induced rise in [Ca²⁺]_i by mGluRs. Parathyroid hormone, which also activates phospholipase C in osteoblasts, had a similar inhibitory effect on the NMDA receptor-induced [Ca²⁺]_i response. Elevation of [Ca²⁺]_i mediated by mGluR activation was reduced by subsequent stimulation of NMDA receptors. This is the first description of mGluRs in bone and shows that complex glutamatergic signaling can occur in this tissue.

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