Accumulation of [3H]glutamate in cultured rat calvarial osteoblasts

Takeshi Takarada, Eiichi Hinoi, Sayumi Fujimori, Yuriko Tsuchihashi, Taichi Ueshima, Hideo Taniura, Yukio Yoneda

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

In the present study, we have attempted to demonstrate constitutive and functional expression in bone of particular glutamate transporters (GluTs) required for signal termination in glutamatergic signaling process. Reverse transcription polymerase chain reaction revealed constitutive expression of mRNA for the neuronal GluT subtype excitatory amino acid carrier-1, in addition to glial subtypes such as glutamate aspartate transporter and glutamate transporter-1, in rat calvarial osteoblasts cultured for 7-21 days in vitro (DIV). The accumulation of [3H]glutamate (Glu) occurred in a temperature- and sodium-dependent manner with pharmacological profiles similar to those for brain GluTs in osteoblasts cultured for 7 DIV, while three different agonists at ionotropic Glu receptors significantly inhibited the accumulation of [3H]Glu in osteoblasts. Although [3H]Glu accumulation consisted of a single component with a Km value of 26.0±5.8 μM and a Vmax value of 960±122 nmol/(min mg protein), respectively, in osteoblasts cultured for 7 DIV, in vitro maturation led to a significant decrease in Vmax value to 290±33 nmol/(min mg protein) without significantly affecting Km values on 21 DIV. These results suggest that Glu could be incorporated into intracellular locations through glial and/or neuronal GluT subtypes expressed in cultured rat calvarial osteoblasts.

Original languageEnglish
Pages (from-to)177-184
Number of pages8
JournalBiochemical Pharmacology
Volume68
Issue number1
DOIs
Publication statusPublished - Jul 1 2004
Externally publishedYes

Keywords

  • DIV
  • EAAC1
  • EAAT
  • GLAST
  • GLT1
  • Glu
  • GluT
  • RT-PCR
  • days in vitro
  • excitatory amino acid carrier-1
  • excitatory amino acid transporter
  • glutamate
  • glutamate aspartate transporter
  • glutamate transporter
  • glutamate transporter-1

ASJC Scopus subject areas

  • Biochemistry
  • Pharmacology

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  • Cite this

    Takarada, T., Hinoi, E., Fujimori, S., Tsuchihashi, Y., Ueshima, T., Taniura, H., & Yoneda, Y. (2004). Accumulation of [3H]glutamate in cultured rat calvarial osteoblasts. Biochemical Pharmacology, 68(1), 177-184. https://doi.org/10.1016/j.bcp.2004.03.020