Pituitary adenylate cyclase-activating polypeptide-induced differentiation of embryonic neural stem cells into astrocytes is mediated via the β isoform of protein kinase C

Jun Watanabe, Motoi Ohba, Fusako Ohno, Sakae Kikuyama, Masahisa Nakamura, Kazuyasu Nakaya, Akira Arimura, Seiji Shioda, Shigeo Nakajo

    研究成果: Article

    22 引用 (Scopus)

    抄録

    We have found previously that pituitary adenylate cyclase-activating polypeptide (PACAP) increases the number of astrocytes generated from cultured mouse neural stem cells (NSCs) via a mechanism that is independent of the cyclic AMP/protein kinase A pathway (Ohno et al., 2005). In the present study, the signaling pathway involved in the differentiation process was further investigated. PACAP-induced differentiation was inhibited by the phospholipase C inhibitor, U73122, the protein kinase C (PKC) inhibitor, chelerythrine, and the intracellular calcium chelator, BAPTA-AM, and was mimicked by phorbol 12-myristate 13-acetate (PMA), but not by 4a-PMA. These results suggest that the PACAP-generated signal was mediated via the PACAP receptor, PAC1 stimulated heterotrimeric G-protein, resulting in activation of phospholipase C, followed by calcium- and phospholipid-dependent protein kinase C (cPKC). To elucidate the involvement of the different isoforms of cPKC, their gene and protein expression were examined. Embryonic NSCs expressed α and βII PKC, but lacked PKCγ. When NSCs were exposed to 2 nM PACAR protein expression levels of the βII isoform transiently increased two-fold before differentiation, returning to basal levels by Day 4, whereas the level of PKCa increased line-arly up to Day 6. Overexpression of PKCβII with adenovirus vector synergistically enhanced differentiation in the presence of 1 nM PACAP, whereas expression of the dominant-negative mutant of PKCβII proved inhibitory. These results indicate that the β isoform of PKC plays a crucial role in the PACAP-induced differentiation of mouse embryonic NSCs into astrocytes.

    元の言語English
    ページ(範囲)1645-1655
    ページ数11
    ジャーナルJournal of Neuroscience Research
    84
    発行部数8
    DOI
    出版物ステータスPublished - 2006 12

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    Pituitary Adenylate Cyclase-Activating Polypeptide
    Neural Stem Cells
    Embryonic Stem Cells
    Astrocytes
    Protein Kinase C
    Protein Isoforms
    Type C Phospholipases
    Acetates
    Pituitary Adenylate Cyclase-Activating Polypeptide Receptors
    Heterotrimeric GTP-Binding Proteins
    Adenylate Kinase
    Protein C Inhibitor
    Protein Kinase Inhibitors
    Cyclic AMP-Dependent Protein Kinases
    Adenoviridae
    Cyclic AMP
    Proteins
    Gene Expression

    ASJC Scopus subject areas

    • Neuroscience(all)

    これを引用

    Pituitary adenylate cyclase-activating polypeptide-induced differentiation of embryonic neural stem cells into astrocytes is mediated via the β isoform of protein kinase C. / Watanabe, Jun; Ohba, Motoi; Ohno, Fusako; Kikuyama, Sakae; Nakamura, Masahisa; Nakaya, Kazuyasu; Arimura, Akira; Shioda, Seiji; Nakajo, Shigeo.

    :: Journal of Neuroscience Research, 巻 84, 番号 8, 12.2006, p. 1645-1655.

    研究成果: Article

    Watanabe, Jun ; Ohba, Motoi ; Ohno, Fusako ; Kikuyama, Sakae ; Nakamura, Masahisa ; Nakaya, Kazuyasu ; Arimura, Akira ; Shioda, Seiji ; Nakajo, Shigeo. / Pituitary adenylate cyclase-activating polypeptide-induced differentiation of embryonic neural stem cells into astrocytes is mediated via the β isoform of protein kinase C. :: Journal of Neuroscience Research. 2006 ; 巻 84, 番号 8. pp. 1645-1655.
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    abstract = "We have found previously that pituitary adenylate cyclase-activating polypeptide (PACAP) increases the number of astrocytes generated from cultured mouse neural stem cells (NSCs) via a mechanism that is independent of the cyclic AMP/protein kinase A pathway (Ohno et al., 2005). In the present study, the signaling pathway involved in the differentiation process was further investigated. PACAP-induced differentiation was inhibited by the phospholipase C inhibitor, U73122, the protein kinase C (PKC) inhibitor, chelerythrine, and the intracellular calcium chelator, BAPTA-AM, and was mimicked by phorbol 12-myristate 13-acetate (PMA), but not by 4a-PMA. These results suggest that the PACAP-generated signal was mediated via the PACAP receptor, PAC1 stimulated heterotrimeric G-protein, resulting in activation of phospholipase C, followed by calcium- and phospholipid-dependent protein kinase C (cPKC). To elucidate the involvement of the different isoforms of cPKC, their gene and protein expression were examined. Embryonic NSCs expressed α and βII PKC, but lacked PKCγ. When NSCs were exposed to 2 nM PACAR protein expression levels of the βII isoform transiently increased two-fold before differentiation, returning to basal levels by Day 4, whereas the level of PKCa increased line-arly up to Day 6. Overexpression of PKCβII with adenovirus vector synergistically enhanced differentiation in the presence of 1 nM PACAP, whereas expression of the dominant-negative mutant of PKCβII proved inhibitory. These results indicate that the β isoform of PKC plays a crucial role in the PACAP-induced differentiation of mouse embryonic NSCs into astrocytes.",
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    T1 - Pituitary adenylate cyclase-activating polypeptide-induced differentiation of embryonic neural stem cells into astrocytes is mediated via the β isoform of protein kinase C

    AU - Watanabe, Jun

    AU - Ohba, Motoi

    AU - Ohno, Fusako

    AU - Kikuyama, Sakae

    AU - Nakamura, Masahisa

    AU - Nakaya, Kazuyasu

    AU - Arimura, Akira

    AU - Shioda, Seiji

    AU - Nakajo, Shigeo

    PY - 2006/12

    Y1 - 2006/12

    N2 - We have found previously that pituitary adenylate cyclase-activating polypeptide (PACAP) increases the number of astrocytes generated from cultured mouse neural stem cells (NSCs) via a mechanism that is independent of the cyclic AMP/protein kinase A pathway (Ohno et al., 2005). In the present study, the signaling pathway involved in the differentiation process was further investigated. PACAP-induced differentiation was inhibited by the phospholipase C inhibitor, U73122, the protein kinase C (PKC) inhibitor, chelerythrine, and the intracellular calcium chelator, BAPTA-AM, and was mimicked by phorbol 12-myristate 13-acetate (PMA), but not by 4a-PMA. These results suggest that the PACAP-generated signal was mediated via the PACAP receptor, PAC1 stimulated heterotrimeric G-protein, resulting in activation of phospholipase C, followed by calcium- and phospholipid-dependent protein kinase C (cPKC). To elucidate the involvement of the different isoforms of cPKC, their gene and protein expression were examined. Embryonic NSCs expressed α and βII PKC, but lacked PKCγ. When NSCs were exposed to 2 nM PACAR protein expression levels of the βII isoform transiently increased two-fold before differentiation, returning to basal levels by Day 4, whereas the level of PKCa increased line-arly up to Day 6. Overexpression of PKCβII with adenovirus vector synergistically enhanced differentiation in the presence of 1 nM PACAP, whereas expression of the dominant-negative mutant of PKCβII proved inhibitory. These results indicate that the β isoform of PKC plays a crucial role in the PACAP-induced differentiation of mouse embryonic NSCs into astrocytes.

    AB - We have found previously that pituitary adenylate cyclase-activating polypeptide (PACAP) increases the number of astrocytes generated from cultured mouse neural stem cells (NSCs) via a mechanism that is independent of the cyclic AMP/protein kinase A pathway (Ohno et al., 2005). In the present study, the signaling pathway involved in the differentiation process was further investigated. PACAP-induced differentiation was inhibited by the phospholipase C inhibitor, U73122, the protein kinase C (PKC) inhibitor, chelerythrine, and the intracellular calcium chelator, BAPTA-AM, and was mimicked by phorbol 12-myristate 13-acetate (PMA), but not by 4a-PMA. These results suggest that the PACAP-generated signal was mediated via the PACAP receptor, PAC1 stimulated heterotrimeric G-protein, resulting in activation of phospholipase C, followed by calcium- and phospholipid-dependent protein kinase C (cPKC). To elucidate the involvement of the different isoforms of cPKC, their gene and protein expression were examined. Embryonic NSCs expressed α and βII PKC, but lacked PKCγ. When NSCs were exposed to 2 nM PACAR protein expression levels of the βII isoform transiently increased two-fold before differentiation, returning to basal levels by Day 4, whereas the level of PKCa increased line-arly up to Day 6. Overexpression of PKCβII with adenovirus vector synergistically enhanced differentiation in the presence of 1 nM PACAP, whereas expression of the dominant-negative mutant of PKCβII proved inhibitory. These results indicate that the β isoform of PKC plays a crucial role in the PACAP-induced differentiation of mouse embryonic NSCs into astrocytes.

    KW - Astrocytes

    KW - G-protein signaling

    KW - PACAP

    KW - PKC

    KW - Stem cells

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