caa a22 4500 475774817 CHVBK 20180406123628.0 cr unu---uuuuu 170329e20000401xx s 000 0 eng 10.1007/s004180000132 doi (NATIONALLICENCE)springer-10.1007/s004180000132 A simple approach for the analysis of intracellular movement of oxidant-producing intracellular compartments in living human neutrophils [Elektronische Daten] [Toshihiro Kobayashi, Vadim S. Zinchuk, Teruhiko Okada, Hiroshi Wakiguchi, Takanobu Kurashige, Hiroyoshi Takatsuji, Harumichi Seguchi] In human neutrophils, superoxide is generated primarily within specialized oxidant-producing intracellular compartments. The present study employs a simple methodological approach to evaluate the intracellular movement of these structures in living human neutrophils. Using a CCD camera system, we monitored fluorescence in cells loaded with the succinimidyl ester of dichlorodihydrofluorescein diacetate, which is non-fluorescent until oxidized by reactive oxygen species. Fluorescence-positive intracellular compartments became detectable after neutrophils were stimulated with phorbol myristate acetate for 1min. Further stimulation increased the intracellular compartments in both number and size in a time-dependent manner. Upon stimulation with phorbol myristate acetate, no fluorescence was seen in intracellular compartments of neutrophils isolated from patients with X-linked chronic granulomatous disease lacking gp91-phox, a membrane component of NADPH oxidase. The method enables tracking of the movement of a single oxidant-producing intracellular compartment following cell stimulation and visualization of the intracellular structures formed by fusion of oxidant-producing intracellular compartments with endocytotic vesicles and phagosomes. Therefore, it is considered to be an informative tool for evaluation of the intracellular dynamics of oxidant-producing intracellular compartments in living human neutrophils and may have a diagnostic value. Springer-Verlag, 2000 Kobayashi Toshihiro Department of Anatomy and Cell Biology, Kochi Medical School, Kohasu, Nankoku-shi, Kochi 783-8505, Japan aut Zinchuk Vadim S. Department of Anatomy and Cell Biology, Kochi Medical School, Kohasu, Nankoku-shi, Kochi 783-8505, Japan aut Okada Teruhiko Department of Anatomy and Cell Biology, Kochi Medical School, Kohasu, Nankoku-shi, Kochi 783-8505, Japan aut Wakiguchi Hiroshi Department of Pediatrics, Kochi Medical School, Kochi 783-8505, Japan aut Kurashige Takanobu Department of Pediatrics, Kochi Medical School, Kochi 783-8505, Japan aut Takatsuji Hiroyoshi Medical Research Center, Kochi Medical School, Kochi 783-8505, Japan aut Seguchi Harumichi Department of Anatomy and Cell Biology, Kochi Medical School, Kohasu, Nankoku-shi, Kochi 783-8505, Japan aut Histochemistry and Cell Biology Springer-Verlag 113/4(2000-04-01), 251-257 0948-6143 113:4<251 2000 113 418 https://doi.org/10.1007/s004180000132 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s004180000132 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kobayashi Toshihiro Department of Anatomy and Cell Biology, Kochi Medical School, Kohasu, Nankoku-shi, Kochi 783-8505, Japan aut NATIONALLICENCE 700 1- Zinchuk Vadim S. Department of Anatomy and Cell Biology, Kochi Medical School, Kohasu, Nankoku-shi, Kochi 783-8505, Japan aut NATIONALLICENCE 700 1- Okada Teruhiko Department of Anatomy and Cell Biology, Kochi Medical School, Kohasu, Nankoku-shi, Kochi 783-8505, Japan aut NATIONALLICENCE 700 1- Wakiguchi Hiroshi Department of Pediatrics, Kochi Medical School, Kochi 783-8505, Japan aut NATIONALLICENCE 700 1- Kurashige Takanobu Department of Pediatrics, Kochi Medical School, Kochi 783-8505, Japan aut NATIONALLICENCE 700 1- Takatsuji Hiroyoshi Medical Research Center, Kochi Medical School, Kochi 783-8505, Japan aut NATIONALLICENCE 700 1- Seguchi Harumichi Department of Anatomy and Cell Biology, Kochi Medical School, Kohasu, Nankoku-shi, Kochi 783-8505, Japan aut NATIONALLICENCE 773 0- Histochemistry and Cell Biology Springer-Verlag 113/4(2000-04-01), 251-257 0948-6143 113:4<251 2000 113 418 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer