Interference with cellular differentiation by D-serine through antagonism at N-methyl-D-aspartate receptors composed of NR1 and NR3A subunits in chondrocytes

Takeshi Takarada, Yoshifumi Takahata, Mika Iemata, Eiichi Hinoi, Kyosuke Uno, Takao Hirai, Tomomi Yamamoto, Yukio Yoneda

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Serine racemase (SR) is responsible for the biosynthesis of D-serine (D-Ser), an endogenous co-agonist for the glycine (Gly)-binding site on N-methyl-D-aspartate (NMDA) receptors, from L-Ser in the brain. We have previously demonstrated high expression of SR by chondrocytes in cartilage. In this study, we attempted to elucidate the possible functional role of D-Ser in chondrogenesis. Expression of mRNA and corresponding protein was seen for SR in cultured rat costal chondrocytes, while the addition of L-Ser significantly increased intracellular and extracellular levels of D-Ser. In organotypic cultured mouse embryonic metatarsals isolated before vascularization, SR mRNA was highly localized in hypertrophic and calcified chondrocytes. Exposure to D-Ser not only suppressed several chondrocytic maturation markers, including alkaline phosphatase (ALP) activity, Ca2+ accumulation, nodule formation, and osteopontin expression, in rat chondrocytes, but also delayed chondral mineralization in mouse metatarsals. Either NMDA or Gly alone significantly increased Ca2+ accumulation in cultured chondrocytes, whereas D-Ser significantly prevented Ca2+ accumulation by Gly, but not by NMDA. Gly alone also significantly increased gene transactivation by the introduction of runt-related transcription factor-2 (Runx2) in COS7 cells transfected with NR1 and NR3A subunits, while D-Ser significantly prevented the increase by Gly without affecting the promoter activity of Runx2. In both cultured chondrocytes and metatarsals from NR1-null mice, significant decreases were seen in ALP activity and chondral mineralization, respectively. These results suggest that D-Ser may negatively regulate cellular differentiation through inhibiting NMDA receptors composed of NR1 and NR3A subunits in a manner related to Runx2 transcriptional activity in chondrocytes.

Original languageEnglish
Pages (from-to)756-764
Number of pages9
JournalJournal of Cellular Physiology
Volume220
Issue number3
DOIs
Publication statusPublished - Sep 2009
Externally publishedYes

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Chondrocytes
N-Methyl-D-Aspartate Receptors
Glycine
Serine
Metatarsal Bones
N-Methylaspartate
Cartilage
Alkaline Phosphatase
Rats
Messenger RNA
Osteopontin
Biosynthesis
Sarcosine
Chondrogenesis
Brain
Transcription Factors
Genes
Binding Sites
Transcriptional Activation
serine racemase

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Interference with cellular differentiation by D-serine through antagonism at N-methyl-D-aspartate receptors composed of NR1 and NR3A subunits in chondrocytes. / Takarada, Takeshi; Takahata, Yoshifumi; Iemata, Mika; Hinoi, Eiichi; Uno, Kyosuke; Hirai, Takao; Yamamoto, Tomomi; Yoneda, Yukio.

In: Journal of Cellular Physiology, Vol. 220, No. 3, 09.2009, p. 756-764.

Research output: Contribution to journalArticle

Takarada, Takeshi ; Takahata, Yoshifumi ; Iemata, Mika ; Hinoi, Eiichi ; Uno, Kyosuke ; Hirai, Takao ; Yamamoto, Tomomi ; Yoneda, Yukio. / Interference with cellular differentiation by D-serine through antagonism at N-methyl-D-aspartate receptors composed of NR1 and NR3A subunits in chondrocytes. In: Journal of Cellular Physiology. 2009 ; Vol. 220, No. 3. pp. 756-764.
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