title: Effective Cellular Morphology Analysis for Differentiation Processes by a Fluorescent 1,3a,6a-Triazapentalene Derivative Probe in Live Cells.
abstract: Nuclear and cytoplasmic morphological changes provide important information about cell differentiation processes, cell functions, and signal responses. There is a strong desire to develop a rapid and simple method for visualizing cytoplasmic and nuclear morphology. Here, we developed a novel and rapid method for probing cellular morphological changes of live cell differentiation process by a fluorescent probe, TAP-4PH, a 1,3a,6a-triazapentalene derivative. TAP-4PH showed high fluorescence in cytoplasmic area, and visualized cytoplasmic and nuclear morphological changes of live cells during differentiation. We demonstrated that TAP-4PH visualized dendritic axon and spine formation in neuronal differentiation, and nuclear structural changes during neutrophilic differentiation. We also showed that the utility of TAP-4PH for visualization of cytoplasmic and nuclear morphologies of various type of live cells. Our visualizing method has no toxicity and no influence on the cellular differentiation and function. The cell morphology can be rapidly observed after addition of TAP-4PH and can continue to be observed in the presence of TAP-4PH in cell culture medium. Moreover, TAP-4PH can be easily removed after observation by washing for subsequent biological assay. Taken together, these results demonstrate that our visualization method is a powerful tool to probe differentiation processes before subsequent biological assay in live cells.
authors.affiliation: Laboratory of Biological Chemistry, Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo, Japan.
authors.forename: Rui
authors.initials: R
authors.lastname: Kamada
authors.affiliation: Laboratory of Biological Chemistry, Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo, Japan.
authors.forename: Fumi
authors.initials: F
authors.lastname: Tano
authors.affiliation: Laboratory of Biological Chemistry, Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo, Japan.
authors.forename: Fuki
authors.initials: F
authors.lastname: Kudoh
authors.affiliation: Laboratory of Biological Chemistry, Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo, Japan.
authors.forename: Nozomi
authors.initials: N
authors.lastname: Kimura
authors.affiliation: Laboratory of Biological Chemistry, Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo, Japan.
authors.forename: Yoshiro
authors.initials: Y
authors.lastname: Chuman
authors.affiliation: Laboratory of Organic Chemistry II, Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo, Japan.
authors.forename: Ayumi
authors.initials: A
authors.lastname: Osawa
authors.affiliation: Graduate School of Pharmaceutical Science, The University of Tokushima, Tokushima, Japan.
authors.forename: Kosuke
authors.initials: K
authors.lastname: Namba
authors.affiliation: Laboratory of Organic Chemistry II, Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo, Japan.
authors.forename: Keiji
authors.initials: K
authors.lastname: Tanino
authors.affiliation: Laboratory of Biological Chemistry, Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo, Japan.
authors.forename: Kazuyasu
authors.initials: K
authors.lastname: Sakaguchi
mesh_terms: D041721:3T3-L1 Cells; D000072283:A549 Cells; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000737:chemistry*; D002453:Cell Cycle; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism*; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D016716:PC12 Cells; D010587:Phagocytosis; D051381:Rats; D013050:Spectrometry, Fluorescence
chemical_list: C568178:1,3a,6a-triazapentalene; D019086:Bridged Bicyclo Compounds, Heterocyclic; D005456:Fluorescent Dyes