Phosphorylation of AMPA-Type Glutamate Receptors by Calcium/ Calmodulin-Dependent Protein Kinase II and Protein Kinase C in Cultured Hippocampal Neurons

Soon Eng Tan, Robert J. Wenthold, Thomas Soderling

    Research output: Contribution to journalArticle

    161 Citations (Scopus)

    Abstract

    Phosphorylation of glutamate receptors (GluRs) is emerging as an important regulatory mechanism. In this study 32P la- beling of non-NMDA GluRs was investigated in cultured hippocampal neurons stimulated 2-15 min with agonists that selectively stimulate either Ca2+/calmodulin-dependent protein kinase II (CaM-kinase II), Ca2+/phospholipid-dependent protein kinase C (PKC), or cAMP-dependent protein kinase A (PKA). Treatment of hippocampal neurons with glutamate/ glycine (Glu/Gly), ionomycin, or 12-O-tetradecanoylphorbol 13-acetate (TPA) increased 32P labeling of immunoprecipitated α-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA)-type GluRs by 145%, 180%, and 227%, respectively, of control values. This increased phosphorylation of GluRs was predominantly 32P-Ser with little 32P-Thr and no detectable 32P-Tyr. Glu/Gly and ionomycin, but not TPA, also increased 32P labeling of CaM-kinase II by 175% and 195%, respectively, of control values. Of these three agonists, only TPA stimulated phosphorylation of MARCKS (225% of control), a specific substrate of PKC. Forskolin treatment gave a three- to fourfold increase in the active catalytic subunit of PKA but did not result in the 32P labeling of AMPA-type GluRs, CaM-kinase II, or MARCKS. Phosphorylation of GluRs in response to Glu/Gly was blocked by a specific NMDA receptor/ion channel antagonist (DL-2-amino-5-phosphon-ovaleric acid) or by a cell-permeable inhibitor of CaM-kinase II (1-[N,O-bis(1,5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine, KN-62). These results are consistent with the hypothesis that Ca2+ influx through the NMDA-type ion channel can activate CaM-kinase II, which in turn can phosphorylate and regulate AMPA-type GluR ion channels (McGlade-McCulloh et al., 1993). Such a mechanism could contribute to the postsynaptic component of long-term potentiation and other forms of synaptic plasticity.

    Original languageEnglish (US)
    Pages (from-to)1123-1129
    Number of pages7
    JournalJournal of Neuroscience
    Volume14
    Issue number3 I
    StatePublished - Mar 1994

    Fingerprint

    Calcium-Calmodulin-Dependent Protein Kinase Kinase
    Calcium-Calmodulin-Dependent Protein Kinase Type 2
    alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid
    Glutamate Receptors
    Protein Kinase C
    KN 62
    Phosphorylation
    Neurons
    Tetradecanoylphorbol Acetate
    Ion Channels
    Glycine
    Glutamic Acid
    Ionomycin
    AMPA Receptors
    Cyclic AMP-Dependent Protein Kinases
    Cyclic AMP-Dependent Protein Kinase Catalytic Subunits
    Neuronal Plasticity
    Long-Term Potentiation
    Colforsin
    N-Methylaspartate

    Keywords

    • Calmodulin-kinase
    • Glutamate receptor
    • Hippocampus
    • Protein kinase
    • Synaptic plasticity

    ASJC Scopus subject areas

    • Neuroscience(all)

    Cite this

    Phosphorylation of AMPA-Type Glutamate Receptors by Calcium/ Calmodulin-Dependent Protein Kinase II and Protein Kinase C in Cultured Hippocampal Neurons. / Tan, Soon Eng; Wenthold, Robert J.; Soderling, Thomas.

    In: Journal of Neuroscience, Vol. 14, No. 3 I, 03.1994, p. 1123-1129.

    Research output: Contribution to journalArticle

    @article{d40d1cd4a1614472842efc3a5409e09c,
    title = "Phosphorylation of AMPA-Type Glutamate Receptors by Calcium/ Calmodulin-Dependent Protein Kinase II and Protein Kinase C in Cultured Hippocampal Neurons",
    abstract = "Phosphorylation of glutamate receptors (GluRs) is emerging as an important regulatory mechanism. In this study 32P la- beling of non-NMDA GluRs was investigated in cultured hippocampal neurons stimulated 2-15 min with agonists that selectively stimulate either Ca2+/calmodulin-dependent protein kinase II (CaM-kinase II), Ca2+/phospholipid-dependent protein kinase C (PKC), or cAMP-dependent protein kinase A (PKA). Treatment of hippocampal neurons with glutamate/ glycine (Glu/Gly), ionomycin, or 12-O-tetradecanoylphorbol 13-acetate (TPA) increased 32P labeling of immunoprecipitated α-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA)-type GluRs by 145{\%}, 180{\%}, and 227{\%}, respectively, of control values. This increased phosphorylation of GluRs was predominantly 32P-Ser with little 32P-Thr and no detectable 32P-Tyr. Glu/Gly and ionomycin, but not TPA, also increased 32P labeling of CaM-kinase II by 175{\%} and 195{\%}, respectively, of control values. Of these three agonists, only TPA stimulated phosphorylation of MARCKS (225{\%} of control), a specific substrate of PKC. Forskolin treatment gave a three- to fourfold increase in the active catalytic subunit of PKA but did not result in the 32P labeling of AMPA-type GluRs, CaM-kinase II, or MARCKS. Phosphorylation of GluRs in response to Glu/Gly was blocked by a specific NMDA receptor/ion channel antagonist (DL-2-amino-5-phosphon-ovaleric acid) or by a cell-permeable inhibitor of CaM-kinase II (1-[N,O-bis(1,5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine, KN-62). These results are consistent with the hypothesis that Ca2+ influx through the NMDA-type ion channel can activate CaM-kinase II, which in turn can phosphorylate and regulate AMPA-type GluR ion channels (McGlade-McCulloh et al., 1993). Such a mechanism could contribute to the postsynaptic component of long-term potentiation and other forms of synaptic plasticity.",
    keywords = "Calmodulin-kinase, Glutamate receptor, Hippocampus, Protein kinase, Synaptic plasticity",
    author = "Tan, {Soon Eng} and Wenthold, {Robert J.} and Thomas Soderling",
    year = "1994",
    month = "3",
    language = "English (US)",
    volume = "14",
    pages = "1123--1129",
    journal = "Journal of Neuroscience",
    issn = "0270-6474",
    publisher = "Society for Neuroscience",
    number = "3 I",

    }

    TY - JOUR

    T1 - Phosphorylation of AMPA-Type Glutamate Receptors by Calcium/ Calmodulin-Dependent Protein Kinase II and Protein Kinase C in Cultured Hippocampal Neurons

    AU - Tan, Soon Eng

    AU - Wenthold, Robert J.

    AU - Soderling, Thomas

    PY - 1994/3

    Y1 - 1994/3

    N2 - Phosphorylation of glutamate receptors (GluRs) is emerging as an important regulatory mechanism. In this study 32P la- beling of non-NMDA GluRs was investigated in cultured hippocampal neurons stimulated 2-15 min with agonists that selectively stimulate either Ca2+/calmodulin-dependent protein kinase II (CaM-kinase II), Ca2+/phospholipid-dependent protein kinase C (PKC), or cAMP-dependent protein kinase A (PKA). Treatment of hippocampal neurons with glutamate/ glycine (Glu/Gly), ionomycin, or 12-O-tetradecanoylphorbol 13-acetate (TPA) increased 32P labeling of immunoprecipitated α-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA)-type GluRs by 145%, 180%, and 227%, respectively, of control values. This increased phosphorylation of GluRs was predominantly 32P-Ser with little 32P-Thr and no detectable 32P-Tyr. Glu/Gly and ionomycin, but not TPA, also increased 32P labeling of CaM-kinase II by 175% and 195%, respectively, of control values. Of these three agonists, only TPA stimulated phosphorylation of MARCKS (225% of control), a specific substrate of PKC. Forskolin treatment gave a three- to fourfold increase in the active catalytic subunit of PKA but did not result in the 32P labeling of AMPA-type GluRs, CaM-kinase II, or MARCKS. Phosphorylation of GluRs in response to Glu/Gly was blocked by a specific NMDA receptor/ion channel antagonist (DL-2-amino-5-phosphon-ovaleric acid) or by a cell-permeable inhibitor of CaM-kinase II (1-[N,O-bis(1,5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine, KN-62). These results are consistent with the hypothesis that Ca2+ influx through the NMDA-type ion channel can activate CaM-kinase II, which in turn can phosphorylate and regulate AMPA-type GluR ion channels (McGlade-McCulloh et al., 1993). Such a mechanism could contribute to the postsynaptic component of long-term potentiation and other forms of synaptic plasticity.

    AB - Phosphorylation of glutamate receptors (GluRs) is emerging as an important regulatory mechanism. In this study 32P la- beling of non-NMDA GluRs was investigated in cultured hippocampal neurons stimulated 2-15 min with agonists that selectively stimulate either Ca2+/calmodulin-dependent protein kinase II (CaM-kinase II), Ca2+/phospholipid-dependent protein kinase C (PKC), or cAMP-dependent protein kinase A (PKA). Treatment of hippocampal neurons with glutamate/ glycine (Glu/Gly), ionomycin, or 12-O-tetradecanoylphorbol 13-acetate (TPA) increased 32P labeling of immunoprecipitated α-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA)-type GluRs by 145%, 180%, and 227%, respectively, of control values. This increased phosphorylation of GluRs was predominantly 32P-Ser with little 32P-Thr and no detectable 32P-Tyr. Glu/Gly and ionomycin, but not TPA, also increased 32P labeling of CaM-kinase II by 175% and 195%, respectively, of control values. Of these three agonists, only TPA stimulated phosphorylation of MARCKS (225% of control), a specific substrate of PKC. Forskolin treatment gave a three- to fourfold increase in the active catalytic subunit of PKA but did not result in the 32P labeling of AMPA-type GluRs, CaM-kinase II, or MARCKS. Phosphorylation of GluRs in response to Glu/Gly was blocked by a specific NMDA receptor/ion channel antagonist (DL-2-amino-5-phosphon-ovaleric acid) or by a cell-permeable inhibitor of CaM-kinase II (1-[N,O-bis(1,5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine, KN-62). These results are consistent with the hypothesis that Ca2+ influx through the NMDA-type ion channel can activate CaM-kinase II, which in turn can phosphorylate and regulate AMPA-type GluR ion channels (McGlade-McCulloh et al., 1993). Such a mechanism could contribute to the postsynaptic component of long-term potentiation and other forms of synaptic plasticity.

    KW - Calmodulin-kinase

    KW - Glutamate receptor

    KW - Hippocampus

    KW - Protein kinase

    KW - Synaptic plasticity

    UR - http://www.scopus.com/inward/record.url?scp=0028218266&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0028218266&partnerID=8YFLogxK

    M3 - Article

    VL - 14

    SP - 1123

    EP - 1129

    JO - Journal of Neuroscience

    JF - Journal of Neuroscience

    SN - 0270-6474

    IS - 3 I

    ER -