TY - JOUR
T1 - Spatiotemporal laser inactivation of inositol 1,4,5-trisphosphate receptors using synthetic small-molecule probes
AU - Inoue, Takanari
AU - Kikuchi, Kazuya
AU - Hirose, Kenzo
AU - Iino, Masamitsu
AU - Nagano, Tetsuo
N1 - Funding Information:
This study was supported in part by the Ministry of Education, Culture, Sports, Science and Technology of Japan (Advanced and Innovational Research Program in Life Sciences and grants 13024217, 13558078, 1367232, and 14045210 to K.K.), Takeda Science Foundation, Nagase Science and Technology Foundation, and Sankyo Foundation. T.I. is a recipient of a fellowship for young scientists from the Japanese Society for the Promotion of Science.
PY - 2003/6/1
Y1 - 2003/6/1
N2 - A malachite green-conjugated inositol 1,4,5-trisphosphate (MGIP3) induces specific inactivation of IP3 receptor (IP3R) in tissue samples upon laser irradiation. To verify potential usefulness of the method for studies of cellular Ca2+ signaling, we conducted laser inactivation at the single-cell level and show that IP3R was inactivated with extremely high spatiotemporal resolution. In the presence of MGIP3, the Ca2+ release function of IP3R in single B lymphoma cells decayed exponentially with increasing duration of laser irradiation with a time constant of 3.4 s. Moreover, by confining laser irradiation to a spatially distinct region of differentiated PC12 cells, subcellular inactivation of IP3R was attained, as revealed by a loss of local Ca2+ signal. Such real-time inactivation of IP3R only within a subcellular region may provide a powerful method for investigating spatiotemporal dynamics of Ca2+ signaling.
AB - A malachite green-conjugated inositol 1,4,5-trisphosphate (MGIP3) induces specific inactivation of IP3 receptor (IP3R) in tissue samples upon laser irradiation. To verify potential usefulness of the method for studies of cellular Ca2+ signaling, we conducted laser inactivation at the single-cell level and show that IP3R was inactivated with extremely high spatiotemporal resolution. In the presence of MGIP3, the Ca2+ release function of IP3R in single B lymphoma cells decayed exponentially with increasing duration of laser irradiation with a time constant of 3.4 s. Moreover, by confining laser irradiation to a spatially distinct region of differentiated PC12 cells, subcellular inactivation of IP3R was attained, as revealed by a loss of local Ca2+ signal. Such real-time inactivation of IP3R only within a subcellular region may provide a powerful method for investigating spatiotemporal dynamics of Ca2+ signaling.
UR - http://www.scopus.com/inward/record.url?scp=0037829824&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037829824&partnerID=8YFLogxK
U2 - 10.1016/S1074-5521(03)00122-4
DO - 10.1016/S1074-5521(03)00122-4
M3 - Article
C2 - 12837383
AN - SCOPUS:0037829824
SN - 1074-5521
VL - 10
SP - 503
EP - 509
JO - Chemistry and Biology
JF - Chemistry and Biology
IS - 6
ER -