Hydrogen-Deuterium Exchange Effects on β-Endorphin Release from AtT20 Murine Pituitary Tumor Cells

Masayuki Ikeda, Shigeru Suzuki, Masahiro Kishio, Moritoshi Hirono, Takashi Sugiyama, Junko Matsuura, Teppei Suzuki, Takayuki Sota, Charles N. Allen, Shiro Konishi, Tohru Yoshioka

    Research output: Contribution to journalArticle

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    Abstract

    Abundant evidences demonstrate that deuterium oxide (D2O) modulates various secretory activities, but specific mechanisms remain unclear. Using AtT20 cells, we examined effects of D2O on physiological processes underlying β-endorphin release. Immunofluorescent confocal microscopy demonstrated that 90% D2O buffer increased the amount of actin filament in cell somas and decreased it in cell processes, whereas β-tubulin was not affected. Ca2+ imaging demonstrated that high-K+-induced Ca2+ influx was not affected during D 2O treatment, but was completely inhibited upon D2O washout. The H2O/D2O replacement in internal solutions of patch electrodes reduced Ca2+ currents evoked by depolarizing voltage steps, whereas additional extracellular H2O/D2O replacement recovered the currents, suggesting that D2O gradient across plasma membrane is critical for Ca2+ channel kinetics. Radioimmunoassay of high-K+-induced β-endorphin release demonstrated an increase during D2O treatment and a decrease upon D2O washout. These results demonstrate that the H 2O-to-D2O-induced increase in β-endorphin release corresponded with the redistribution of actin, and the D2O-to-H 2O-induced decrease in β-endorphin release corresponded with the inhibition of voltage-sensitive Ca2+ channels. The computer modeling suggests that the differences in the zero-point vibrational energy between protonated and deuterated amino acids produce an asymmetric distribution of these amino acids upon D2O washout and this causes the dysfunction of Ca2+ channels.

    Original languageEnglish
    Pages (from-to)565-575
    Number of pages11
    JournalBiophysical Journal
    Volume86
    Issue number1 I
    Publication statusPublished - 2004 Jan

    Fingerprint

    Endorphins
    Deuterium
    Pituitary Neoplasms
    Hydrogen
    Physiological Phenomena
    Deuterium Oxide
    Amino Acids
    Carisoprodol
    Tubulin
    Actin Cytoskeleton
    Confocal Microscopy
    Radioimmunoassay
    Actins
    Buffers
    Electrodes
    Cell Membrane

    ASJC Scopus subject areas

    • Biophysics

    Cite this

    Ikeda, M., Suzuki, S., Kishio, M., Hirono, M., Sugiyama, T., Matsuura, J., ... Yoshioka, T. (2004). Hydrogen-Deuterium Exchange Effects on β-Endorphin Release from AtT20 Murine Pituitary Tumor Cells. Biophysical Journal, 86(1 I), 565-575.

    Hydrogen-Deuterium Exchange Effects on β-Endorphin Release from AtT20 Murine Pituitary Tumor Cells. / Ikeda, Masayuki; Suzuki, Shigeru; Kishio, Masahiro; Hirono, Moritoshi; Sugiyama, Takashi; Matsuura, Junko; Suzuki, Teppei; Sota, Takayuki; Allen, Charles N.; Konishi, Shiro; Yoshioka, Tohru.

    In: Biophysical Journal, Vol. 86, No. 1 I, 01.2004, p. 565-575.

    Research output: Contribution to journalArticle

    Ikeda, M, Suzuki, S, Kishio, M, Hirono, M, Sugiyama, T, Matsuura, J, Suzuki, T, Sota, T, Allen, CN, Konishi, S & Yoshioka, T 2004, 'Hydrogen-Deuterium Exchange Effects on β-Endorphin Release from AtT20 Murine Pituitary Tumor Cells', Biophysical Journal, vol. 86, no. 1 I, pp. 565-575.
    Ikeda M, Suzuki S, Kishio M, Hirono M, Sugiyama T, Matsuura J et al. Hydrogen-Deuterium Exchange Effects on β-Endorphin Release from AtT20 Murine Pituitary Tumor Cells. Biophysical Journal. 2004 Jan;86(1 I):565-575.
    Ikeda, Masayuki ; Suzuki, Shigeru ; Kishio, Masahiro ; Hirono, Moritoshi ; Sugiyama, Takashi ; Matsuura, Junko ; Suzuki, Teppei ; Sota, Takayuki ; Allen, Charles N. ; Konishi, Shiro ; Yoshioka, Tohru. / Hydrogen-Deuterium Exchange Effects on β-Endorphin Release from AtT20 Murine Pituitary Tumor Cells. In: Biophysical Journal. 2004 ; Vol. 86, No. 1 I. pp. 565-575.
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