Bax shuttling after rotenone treatment of neuronal primary cultures: Effects on cell death phenotypes

Martin B. Gill, J. Regino Perez-Polo

    Research output: Contribution to journalArticle

    13 Citations (Scopus)

    Abstract

    Neonatal (P7) brain hypoxia-ischemia (HI) induces intracellular Bax protein shifts to the nucleus, mitochondria, and endoplasmic reticulum (ER), where it triggers the activation of the respective cell death signaling cascades. When compared with HI-treated rat pups, 100% D2 resuscitation of HI-treated rat pups increases HI-induced ER Bax levels, ER-mediated cell death signaling, and resultant lesion volume and inflammation due to increased necrotic-like cell death. To better characterize the role of Bax intracellular shuttling ER cell death signaling and necrotic-like cell death, we used rotenone-treated P5 neuronal cortical cultures to increase ER Bax levels and subsequent cell death signaling. We treated P5 primary cortical neurons with 25 μM and 100 μM rotenone as an apoptotic or necrotic-like stimulus, respectively, and measured intracellular organelle Bax levels and the subsequent activation of ER/mitochondrial cell death signaling. The 25 μM rotenone treatment promptly increased nuclear Bax levels followed by a later increase in mitochondrial Bax levels and caspase-mediated cleavage of α-fodrin. The 100 μM rotenone treatment also resulted in an early increase in nuclear Bax levels followed by a subsequent increase in ER Bax levels and calpain-mediated cleavage of α-fodrin. After pretreatment with the immunosuppressive and neuroprotective FK506, there was a delay in Bax intracellular shifts and cell death signaling for both the 25 and 100 μM rotenone treatments. These results suggest that the different outcomes of apoptotic-like vs. necrotic-like cell death resulting from the treatment of neuronal cultures with rotenone at 25 and 100 μM rotenone reflect changes in the intracellular trafficking of Bax among different organelles.

    Original languageEnglish (US)
    Pages (from-to)2047-2065
    Number of pages19
    JournalJournal of Neuroscience Research
    Volume87
    Issue number9
    DOIs
    StatePublished - 2009

    Fingerprint

    Rotenone
    Cell Death
    Endoplasmic Reticulum
    Phenotype
    Therapeutics
    Ischemia
    Organelles
    Brain Hypoxia-Ischemia
    bcl-2-Associated X Protein
    Calpain
    Tacrolimus
    Immunosuppressive Agents
    Caspases
    Resuscitation
    Mitochondria
    Inflammation
    Neurons

    Keywords

    • Apoptosis
    • Bax
    • Calpain
    • Necrosis
    • Neuron
    • Rotenone

    ASJC Scopus subject areas

    • Cellular and Molecular Neuroscience

    Cite this

    Bax shuttling after rotenone treatment of neuronal primary cultures : Effects on cell death phenotypes. / Gill, Martin B.; Perez-Polo, J. Regino.

    In: Journal of Neuroscience Research, Vol. 87, No. 9, 2009, p. 2047-2065.

    Research output: Contribution to journalArticle

    @article{5fa32c00038a43a9b5cbf94dddea4b90,
    title = "Bax shuttling after rotenone treatment of neuronal primary cultures: Effects on cell death phenotypes",
    abstract = "Neonatal (P7) brain hypoxia-ischemia (HI) induces intracellular Bax protein shifts to the nucleus, mitochondria, and endoplasmic reticulum (ER), where it triggers the activation of the respective cell death signaling cascades. When compared with HI-treated rat pups, 100{\%} D2 resuscitation of HI-treated rat pups increases HI-induced ER Bax levels, ER-mediated cell death signaling, and resultant lesion volume and inflammation due to increased necrotic-like cell death. To better characterize the role of Bax intracellular shuttling ER cell death signaling and necrotic-like cell death, we used rotenone-treated P5 neuronal cortical cultures to increase ER Bax levels and subsequent cell death signaling. We treated P5 primary cortical neurons with 25 μM and 100 μM rotenone as an apoptotic or necrotic-like stimulus, respectively, and measured intracellular organelle Bax levels and the subsequent activation of ER/mitochondrial cell death signaling. The 25 μM rotenone treatment promptly increased nuclear Bax levels followed by a later increase in mitochondrial Bax levels and caspase-mediated cleavage of α-fodrin. The 100 μM rotenone treatment also resulted in an early increase in nuclear Bax levels followed by a subsequent increase in ER Bax levels and calpain-mediated cleavage of α-fodrin. After pretreatment with the immunosuppressive and neuroprotective FK506, there was a delay in Bax intracellular shifts and cell death signaling for both the 25 and 100 μM rotenone treatments. These results suggest that the different outcomes of apoptotic-like vs. necrotic-like cell death resulting from the treatment of neuronal cultures with rotenone at 25 and 100 μM rotenone reflect changes in the intracellular trafficking of Bax among different organelles.",
    keywords = "Apoptosis, Bax, Calpain, Necrosis, Neuron, Rotenone",
    author = "Gill, {Martin B.} and Perez-Polo, {J. Regino}",
    year = "2009",
    doi = "10.1002/jnr.22019",
    language = "English (US)",
    volume = "87",
    pages = "2047--2065",
    journal = "Journal of Neuroscience Research",
    issn = "0360-4012",
    publisher = "Wiley-Liss Inc.",
    number = "9",

    }

    TY - JOUR

    T1 - Bax shuttling after rotenone treatment of neuronal primary cultures

    T2 - Effects on cell death phenotypes

    AU - Gill, Martin B.

    AU - Perez-Polo, J. Regino

    PY - 2009

    Y1 - 2009

    N2 - Neonatal (P7) brain hypoxia-ischemia (HI) induces intracellular Bax protein shifts to the nucleus, mitochondria, and endoplasmic reticulum (ER), where it triggers the activation of the respective cell death signaling cascades. When compared with HI-treated rat pups, 100% D2 resuscitation of HI-treated rat pups increases HI-induced ER Bax levels, ER-mediated cell death signaling, and resultant lesion volume and inflammation due to increased necrotic-like cell death. To better characterize the role of Bax intracellular shuttling ER cell death signaling and necrotic-like cell death, we used rotenone-treated P5 neuronal cortical cultures to increase ER Bax levels and subsequent cell death signaling. We treated P5 primary cortical neurons with 25 μM and 100 μM rotenone as an apoptotic or necrotic-like stimulus, respectively, and measured intracellular organelle Bax levels and the subsequent activation of ER/mitochondrial cell death signaling. The 25 μM rotenone treatment promptly increased nuclear Bax levels followed by a later increase in mitochondrial Bax levels and caspase-mediated cleavage of α-fodrin. The 100 μM rotenone treatment also resulted in an early increase in nuclear Bax levels followed by a subsequent increase in ER Bax levels and calpain-mediated cleavage of α-fodrin. After pretreatment with the immunosuppressive and neuroprotective FK506, there was a delay in Bax intracellular shifts and cell death signaling for both the 25 and 100 μM rotenone treatments. These results suggest that the different outcomes of apoptotic-like vs. necrotic-like cell death resulting from the treatment of neuronal cultures with rotenone at 25 and 100 μM rotenone reflect changes in the intracellular trafficking of Bax among different organelles.

    AB - Neonatal (P7) brain hypoxia-ischemia (HI) induces intracellular Bax protein shifts to the nucleus, mitochondria, and endoplasmic reticulum (ER), where it triggers the activation of the respective cell death signaling cascades. When compared with HI-treated rat pups, 100% D2 resuscitation of HI-treated rat pups increases HI-induced ER Bax levels, ER-mediated cell death signaling, and resultant lesion volume and inflammation due to increased necrotic-like cell death. To better characterize the role of Bax intracellular shuttling ER cell death signaling and necrotic-like cell death, we used rotenone-treated P5 neuronal cortical cultures to increase ER Bax levels and subsequent cell death signaling. We treated P5 primary cortical neurons with 25 μM and 100 μM rotenone as an apoptotic or necrotic-like stimulus, respectively, and measured intracellular organelle Bax levels and the subsequent activation of ER/mitochondrial cell death signaling. The 25 μM rotenone treatment promptly increased nuclear Bax levels followed by a later increase in mitochondrial Bax levels and caspase-mediated cleavage of α-fodrin. The 100 μM rotenone treatment also resulted in an early increase in nuclear Bax levels followed by a subsequent increase in ER Bax levels and calpain-mediated cleavage of α-fodrin. After pretreatment with the immunosuppressive and neuroprotective FK506, there was a delay in Bax intracellular shifts and cell death signaling for both the 25 and 100 μM rotenone treatments. These results suggest that the different outcomes of apoptotic-like vs. necrotic-like cell death resulting from the treatment of neuronal cultures with rotenone at 25 and 100 μM rotenone reflect changes in the intracellular trafficking of Bax among different organelles.

    KW - Apoptosis

    KW - Bax

    KW - Calpain

    KW - Necrosis

    KW - Neuron

    KW - Rotenone

    UR - http://www.scopus.com/inward/record.url?scp=65849142691&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=65849142691&partnerID=8YFLogxK

    U2 - 10.1002/jnr.22019

    DO - 10.1002/jnr.22019

    M3 - Article

    C2 - 19224578

    AN - SCOPUS:65849142691

    VL - 87

    SP - 2047

    EP - 2065

    JO - Journal of Neuroscience Research

    JF - Journal of Neuroscience Research

    SN - 0360-4012

    IS - 9

    ER -