Roles of ionotropic glutamate receptors in early developing neurons derived from the P19 mouse cell line

Yi-Hsuan Lee, Chun Hua Lin, Li Wen Hsu, Ssu Yao Hu, Wen Te Hsiao, Yuan Soon Ho

Research output: Contribution to journalArticle

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Abstract

We cultured a P19 mouse teratocarcinoma cell line and induced its neuronal differentiation to study the function of ionotropic glutamate receptors (GluRs) in early neuronal development. Immunocytochemical studies showed 85% neuronal population at 5 days in vitro (DIV) with microtubule-associated protein 2-positive staining. Thirty percent and 50% of the cells expressed the α-amino-3-hydroxy-5-methyl-4-isopropinonate (AMPA) receptor subunit, GluR2/3, and the kainate (kainic acid; KA) receptor subunit, GluR5/6/7, respectively. In Western blot analysis, the temporal expression of GluR2/3 began to appear at 3 DIV, whereas GluR5/6/7 was already expressed in the undifferentiated cells. P19-derived neurons began to respond to glutamate, AMPA and KA, but not to the metabotropic GluR agonist trans-1-aminocyclopentane-1,3-decarboxylic acid, by 5 DIV in terms of increases in intracellular calcium and phospholipase C-mediated poly-phosphoinositide turnover. Furthermore, KA reduced cell death of P19-derived neurons in both atmospheric and hypobaric conditions in a phospholipase C-dependent manner. The common AMPA/KA receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione, but not the AMPA receptor antagonist, 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo-benzo[f]quinoxaline-7-sulfonamide disodium, profoundly increased hypobaric insult-induced neurotoxicity. In a flow cytometry study, the nerve growth factor-mediated antiapoptotic effect was facilitated by AMPA, with an induction of TrkA, but not p75NTR expression. Therefore, AMPA and KA receptors might mediate neurotrophic functions to facilitate neurotrophic factor signaling to protect neurons against hypoxic insult in early neuronal development.

Original languageEnglish
Pages (from-to)199-207
Number of pages9
JournalJournal of Biomedical Science
Volume10
Issue number2
DOIs
Publication statusPublished - 2003

Fingerprint

Ionotropic Glutamate Receptors
alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid
AMPA Receptors
Neurons
Cells
Type C Phospholipases
Cell Line
Kainic Acid
Teratocarcinoma
6-Cyano-7-nitroquinoxaline-2,3-dione
Kainic Acid Receptors
Microtubule-Associated Proteins
Sulfonamides
Nerve Growth Factors
Nerve Growth Factor
Phosphatidylinositols
Flow cytometry
Glutamic Acid
Flow Cytometry
Cell Death

Keywords

  • Glutamate receptor
  • Hypoxia
  • Kainic acid
  • Nerve growth factor
  • P19 cells

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Roles of ionotropic glutamate receptors in early developing neurons derived from the P19 mouse cell line. / Lee, Yi-Hsuan; Lin, Chun Hua; Hsu, Li Wen; Hu, Ssu Yao; Hsiao, Wen Te; Ho, Yuan Soon.

In: Journal of Biomedical Science, Vol. 10, No. 2, 2003, p. 199-207.

Research output: Contribution to journalArticle

Lee, Yi-Hsuan ; Lin, Chun Hua ; Hsu, Li Wen ; Hu, Ssu Yao ; Hsiao, Wen Te ; Ho, Yuan Soon. / Roles of ionotropic glutamate receptors in early developing neurons derived from the P19 mouse cell line. In: Journal of Biomedical Science. 2003 ; Vol. 10, No. 2. pp. 199-207.
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AB - We cultured a P19 mouse teratocarcinoma cell line and induced its neuronal differentiation to study the function of ionotropic glutamate receptors (GluRs) in early neuronal development. Immunocytochemical studies showed 85% neuronal population at 5 days in vitro (DIV) with microtubule-associated protein 2-positive staining. Thirty percent and 50% of the cells expressed the α-amino-3-hydroxy-5-methyl-4-isopropinonate (AMPA) receptor subunit, GluR2/3, and the kainate (kainic acid; KA) receptor subunit, GluR5/6/7, respectively. In Western blot analysis, the temporal expression of GluR2/3 began to appear at 3 DIV, whereas GluR5/6/7 was already expressed in the undifferentiated cells. P19-derived neurons began to respond to glutamate, AMPA and KA, but not to the metabotropic GluR agonist trans-1-aminocyclopentane-1,3-decarboxylic acid, by 5 DIV in terms of increases in intracellular calcium and phospholipase C-mediated poly-phosphoinositide turnover. Furthermore, KA reduced cell death of P19-derived neurons in both atmospheric and hypobaric conditions in a phospholipase C-dependent manner. The common AMPA/KA receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione, but not the AMPA receptor antagonist, 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo-benzo[f]quinoxaline-7-sulfonamide disodium, profoundly increased hypobaric insult-induced neurotoxicity. In a flow cytometry study, the nerve growth factor-mediated antiapoptotic effect was facilitated by AMPA, with an induction of TrkA, but not p75NTR expression. Therefore, AMPA and KA receptors might mediate neurotrophic functions to facilitate neurotrophic factor signaling to protect neurons against hypoxic insult in early neuronal development.

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