Hydrogen-deuterium exchange effects on beta-endorphin release from AtT20 murine pituitary tumor cells.
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 beta-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 beta-tubulin was not affected. Ca2+ imaging demonstrated that high-K+-induced Ca2+ influx was not affected during D2O 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 beta-endorphin release demonstrated an increase during D2O treatment and a decrease upon D(2)O washout. These results demonstrate that the H2O-to-D2O-induced increase in beta-endorphin release corresponded with the redistribution of actin, and the D2O-to-H2O-induced decrease in beta-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.
Ikeda
Masayuki
M
Advanced Research Institute for Science and Engineering, Waseda University, 3-4-1 Okubo Shinjuku-ku, Tokyo 169-8555, Japan.
Suzuki
Shigeru
S
Kishio
Masahiro
M
Hirono
Moritoshi
M
Sugiyama
Takashi
T
Matsuura
Junko
J
Suzuki
Teppei
T
Sota
Takayuki
T
Allen
Charles N
CN
Konishi
Shiro
S
Yoshioka
Tohru
T
D008841:Actin Cytoskeleton / Q000378:metabolism*; D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects*; D045744:Cell Line, Tumor / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D003198:Computer Simulation; D017666:Deuterium Oxide / Q000494:pharmacology*; D006859:Hydrogen / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D010911:Pituitary Neoplasms / Q000378:metabolism* / Q000473:pathology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014018:Tissue Distribution; D014867:Water / Q000378:metabolism; D001615:beta-Endorphin / Q000378:metabolism*
D015220:Calcium Channels; D014867:Water; D001615:beta-Endorphin; D006859:Hydrogen; D017666:Deuterium Oxide; D002118:Calcium