Molecular identification of NMDA glutamate receptors expressed in bone cells

Cecile Itzstein, H Cheynel, B Burt-Pichat, B Merle, L Espinosa, P D Delmas, C Chenu

Research output: Contribution to journalArticle

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Abstract

The N-methyl-D-aspartate (NMDA) subtype of the glutamate receptor has recently been identified in bone, but the molecular composition of this receptor expressed by bone cells is unknown. NMDA receptor (NMDAR) is a hetero-oligomeric protein composed of two classes of subunits, the essential subunit NR1 and NR2A to D subunits that do not by themselves produce functional channels but potentiate NR1 activity and confer functional variability to the receptor. These subunits coassemble in different combinations to form functionally distinct NMDAR. In this study, we have investigated the molecular composition of NMDAR expressed by osteoblasts and osteoclasts in culture, using RT-PCR analysis, in situ hybridization and immunocytochemistry. Specific probes were designed for the different subunits of the NMDAR, and we showed by RT-PCR analysis that mammalian osteoclasts expressed NR2B and NR2D subunits mRNAs but not NR2A and NR2C mRNAs. Rat calvaria and MG63 osteoblastic cells also expressed several NR2 subunits mRNAs, namely NR2A, NR2B, and NR2D. In situ hybridization on isolated rabbit osteoclasts and MG63 cells has confirmed the localization of NR1, NR2B, and NR2D transcripts in osteoclasts and NR1, NR2A, NR2B, and NR2D transcripts in MG63 cells. The expression of NR2D protein by bone cells was shown by immunofluorescence. These results demonstrate for the first time that osteoblasts and osteoclasts express several NR2 subunits, suggesting a molecular diversity of NMDAR channels similar to what was shown for brain. The presence of distinct functional NMDAR on bone cells may be associated with various states of bone cell differentiation and function.
Original languageEnglish
Pages (from-to)134-44
Number of pages11
JournalJournal of Cellular Biochemistry
Volume82
Issue number1
Publication statusPublished - 2001

Fingerprint

Glutamate Receptors
N-Methylaspartate
N-Methyl-D-Aspartate Receptors
Osteoclasts
Bone
Bone and Bones
Osteoblasts
Messenger RNA
In Situ Hybridization
Polymerase Chain Reaction
Chemical analysis
Cell culture
Skull
Fluorescent Antibody Technique
N-methylglutamate
Rats
Cell Differentiation
Brain
Proteins
Immunohistochemistry

Keywords

  • Animals
  • Bone and Bones
  • Cells, Cultured
  • Fluorescent Antibody Technique
  • Humans
  • In Situ Hybridization
  • Mammals
  • Osteoblasts
  • Osteoclasts
  • Patch-Clamp Techniques
  • Protein Subunits
  • Rabbits
  • Rats
  • Receptors, N-Methyl-D-Aspartate
  • Reverse Transcriptase Polymerase Chain Reaction
  • Skull

Cite this

Itzstein, C., Cheynel, H., Burt-Pichat, B., Merle, B., Espinosa, L., Delmas, P. D., & Chenu, C. (2001). Molecular identification of NMDA glutamate receptors expressed in bone cells. Journal of Cellular Biochemistry, 82(1), 134-44.

Molecular identification of NMDA glutamate receptors expressed in bone cells. / Itzstein, Cecile; Cheynel, H; Burt-Pichat, B; Merle, B; Espinosa, L; Delmas, P D; Chenu, C.

In: Journal of Cellular Biochemistry, Vol. 82, No. 1, 2001, p. 134-44.

Research output: Contribution to journalArticle

Itzstein, C, Cheynel, H, Burt-Pichat, B, Merle, B, Espinosa, L, Delmas, PD & Chenu, C 2001, 'Molecular identification of NMDA glutamate receptors expressed in bone cells', Journal of Cellular Biochemistry, vol. 82, no. 1, pp. 134-44.
Itzstein C, Cheynel H, Burt-Pichat B, Merle B, Espinosa L, Delmas PD et al. Molecular identification of NMDA glutamate receptors expressed in bone cells. Journal of Cellular Biochemistry. 2001;82(1):134-44.
Itzstein, Cecile ; Cheynel, H ; Burt-Pichat, B ; Merle, B ; Espinosa, L ; Delmas, P D ; Chenu, C. / Molecular identification of NMDA glutamate receptors expressed in bone cells. In: Journal of Cellular Biochemistry. 2001 ; Vol. 82, No. 1. pp. 134-44.
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T1 - Molecular identification of NMDA glutamate receptors expressed in bone cells

AU - Itzstein, Cecile

AU - Cheynel, H

AU - Burt-Pichat, B

AU - Merle, B

AU - Espinosa, L

AU - Delmas, P D

AU - Chenu, C

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PY - 2001

Y1 - 2001

N2 - The N-methyl-D-aspartate (NMDA) subtype of the glutamate receptor has recently been identified in bone, but the molecular composition of this receptor expressed by bone cells is unknown. NMDA receptor (NMDAR) is a hetero-oligomeric protein composed of two classes of subunits, the essential subunit NR1 and NR2A to D subunits that do not by themselves produce functional channels but potentiate NR1 activity and confer functional variability to the receptor. These subunits coassemble in different combinations to form functionally distinct NMDAR. In this study, we have investigated the molecular composition of NMDAR expressed by osteoblasts and osteoclasts in culture, using RT-PCR analysis, in situ hybridization and immunocytochemistry. Specific probes were designed for the different subunits of the NMDAR, and we showed by RT-PCR analysis that mammalian osteoclasts expressed NR2B and NR2D subunits mRNAs but not NR2A and NR2C mRNAs. Rat calvaria and MG63 osteoblastic cells also expressed several NR2 subunits mRNAs, namely NR2A, NR2B, and NR2D. In situ hybridization on isolated rabbit osteoclasts and MG63 cells has confirmed the localization of NR1, NR2B, and NR2D transcripts in osteoclasts and NR1, NR2A, NR2B, and NR2D transcripts in MG63 cells. The expression of NR2D protein by bone cells was shown by immunofluorescence. These results demonstrate for the first time that osteoblasts and osteoclasts express several NR2 subunits, suggesting a molecular diversity of NMDAR channels similar to what was shown for brain. The presence of distinct functional NMDAR on bone cells may be associated with various states of bone cell differentiation and function.

AB - The N-methyl-D-aspartate (NMDA) subtype of the glutamate receptor has recently been identified in bone, but the molecular composition of this receptor expressed by bone cells is unknown. NMDA receptor (NMDAR) is a hetero-oligomeric protein composed of two classes of subunits, the essential subunit NR1 and NR2A to D subunits that do not by themselves produce functional channels but potentiate NR1 activity and confer functional variability to the receptor. These subunits coassemble in different combinations to form functionally distinct NMDAR. In this study, we have investigated the molecular composition of NMDAR expressed by osteoblasts and osteoclasts in culture, using RT-PCR analysis, in situ hybridization and immunocytochemistry. Specific probes were designed for the different subunits of the NMDAR, and we showed by RT-PCR analysis that mammalian osteoclasts expressed NR2B and NR2D subunits mRNAs but not NR2A and NR2C mRNAs. Rat calvaria and MG63 osteoblastic cells also expressed several NR2 subunits mRNAs, namely NR2A, NR2B, and NR2D. In situ hybridization on isolated rabbit osteoclasts and MG63 cells has confirmed the localization of NR1, NR2B, and NR2D transcripts in osteoclasts and NR1, NR2A, NR2B, and NR2D transcripts in MG63 cells. The expression of NR2D protein by bone cells was shown by immunofluorescence. These results demonstrate for the first time that osteoblasts and osteoclasts express several NR2 subunits, suggesting a molecular diversity of NMDAR channels similar to what was shown for brain. The presence of distinct functional NMDAR on bone cells may be associated with various states of bone cell differentiation and function.

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KW - Bone and Bones

KW - Cells, Cultured

KW - Fluorescent Antibody Technique

KW - Humans

KW - In Situ Hybridization

KW - Mammals

KW - Osteoblasts

KW - Osteoclasts

KW - Patch-Clamp Techniques

KW - Protein Subunits

KW - Rabbits

KW - Rats

KW - Receptors, N-Methyl-D-Aspartate

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Skull

M3 - Article

VL - 82

SP - 134

EP - 144

JO - Journal of Cellular Biochemistry

JF - Journal of Cellular Biochemistry

SN - 0730-2312

IS - 1

ER -