Optical Bioimaging : From Living Tissue to a Single Molecule : Calcium Imaging in Blood Vessel In Situ Employing Two-Photon Excitation Fluorescence Microscopy
スポンサーリンク
概要
- 論文の詳細を見る
Recent developments in optoelectronics permit real-time Ca2+ imaging of thin planes within cells utilizing laser scanning confocal microscopy (LSCM). However, a major complication associated with this imaging system involves increased phototoxicity with improved spatiotemporal resolution. Two-photon excitation microscopy (TPEM) helps to minimize phototoxicity due to the restriction of this technique to the volume proximal to the geometric focus of the light. In this study, the capability of Ca2+ imaging was investigated employing recently developed real-time TPEM, RTS2000MP (Bio-Rad, Tokyo) with a mode-locked Ti-sapphire laser. Z-axis resolution of RTS2000MP with high NA objectives defined as full-width at half maximum (FWHM) with a 0.5-μm fluorescent bead provided values nearly identical to those obtained with LSCM at a small pinhole (0.2 mm) (approximately 0.6 μm). When serial sectioning of 21 sequential images at 0.3-μm intervals in cultured endothelial cells loaded with calcein and tetramethyl-rhodamine methylester were performed with TPEM, the z-axis resolution was higher than that observed with LSCM; moreover, the photobleaching rate was significantly lower than that obtained with LSCM. Maximum fluorescence intensities were detected at 780 nm in excitation spectra of fluo-3 and fluo-4 Ca2+-sensitive probes with TPEM. Fluorescence images in mouse arterial endothelial cells loaded with fluo-4 could be clearly visualized by TPEM in situ. Application of acetylcholine caused oscillatory increase in [Ca2+]i of endothelial cells; subsequently, relaxation along the major axis of smooth muscle cells was evident. Furthermore, consecutive long-lasting experiments could be repeated with identical response in the same microscopic field. In conclusion, fluorescence imaging employing TPEM is useful for Ca2+ imaging in blood vessels in situ.
- 社団法人 日本薬理学会の論文
- 2003-11-01
著者
-
YAMAMOTO Masayuki
Department of Biochemistry, Tohoku University School of Medicine
-
OHATA Hisayuki
Department of Pharmacology, School of Pharmaceutical Sciences, Showa University
-
MOMOSE Kazutaka
Department of Pharmacology, School of Pharmaceutical Sciences, Showa University
-
Ohata H
Department Of Pharmacology School Of Pharmaceutical Sciences Showa University
-
Ohata Hisayuki
Department Of Pharmacology School Of Pharmaceutical Sciences Showa University
-
Yamamoto M
Department Of Pharmacology School Of Pharmaceutical Sciences Showa University
-
Momose K
Department Of Pharmacology School Of Pharmaceutical Sciences Showa University
-
Momose Kazutaka
Department Of Phamacology School Of Pharmaceutical Sciences Showa University
-
YAMADA Hideyuki
Department of Pharmacology, School of Pharmaceutical Sciences, Showa University
-
NIIOKA Takeharu
Department of Pharmacology, School of Pharmaceutical Sciences, Showa University
-
Yamada Hideyuki
Department Of Ophthalmology Faculty Of Medicine University Of Tokyo
-
Yamada Hideyuki
Department Of Pharmacology School Of Pharmaceutical Sciences Showa University
-
Niioka Takeharu
Department Of Oral Physiology School Of Dentistry Health Sciences University Of Hokkaido
-
Yamamoto Masayuki
Department Of Biochemistry Tohoku University School Of Medicine
-
Yamamoto Masayuki
Department of Anatomy, Hiroshima University school of Medicine
-
YAMADA Hideyuki
Department of Hygienic and Forensic Chemistry, Faculty of Pharmaceutical Sciences
関連論文
- Conductance Distribution in Disordered Quantum Wires with a Perfectly Conducting Channel(Condensed matter: electronic structure and electrical, magnetic, and optical properties)
- Conserved Structure, Regulatory Elements, and Transcriptional Regulation from the GATA-1 Gene Testis Promoter^1
- Role of Rho-Associated Protein Kinase and Histamine in Lysophosphatidic Acid-Induced Airway Hyperresponsiveness in Guinea Pigs
- Effects of Papain on Isolaton of Single Smooth Muscle Cells from the Guinea Pig Longitudinal Ileum
- SPONTANEOUS LUNG METASTATIC MODEL OF LEWIS LUNG CANCER IN MOUSE VISUALIZED BY GREEN FLUORESCENT PROTEIN EXPRESSION
- High-Glucose Enhances a Thromboxane A_2-Induced Aortic Contraction Mediated by an Alteration of Phosphatidylinositol Turnover
- Role of Rho-Associated Protein Kinase and Histamine in Lysophosphatidic Acid-Induced Airway Hyperresponsiveness in Guinea Pigs
- Characterization of the C. elegans homolog of Mblk-1 encoding a putative transcription factor that is expressed in the honey bee brain(Physiology)(Proceeding of the Seventy-Third Annual Meeting of the Zoological Society of Japan)
- CYTOTOXICITY AND SUPPRESSION OF CALCIUM RESPONSE BY TRIBUTYLTIN-TREATMENT IN PRIMARY CULTURED HEPATOCYTES
- Suppression of Calcium Response to Hormonal Stimulation by Tributyltin in Primary Cultured Rat Hepatocytes
- Caspase Cascade Proceeds Rapidly After Cytochrome c Release From Mitochondria in Tumor Necrosis Factor-α-Induced Cell Death
- Simultaneous Real-Time Detection of Initiator- and Effector-Caspase Activation by Double Fluorescence Resonance Energy Transfer Analysis
- Cyst Fluid Levels of Carcinoembryonic Antigen, Carbohydrate Antigen 19-9, Squamous Cell Carcinoma Antigen, and Amylase in Thyroglossal Duct and Branchial Cleft Cysts
- Detection and characterization of caffeine-less tea plants originated from interspecific hybridization
- Siblings with Malignant Glioma : Report of Two Cases
- Purification and Characterization of a Major Collagenase from Streptomyces parvulus
- Detection of Carcinoembryonic Antigen mRNA in the Mesenteric Vein of Patients with Resectable Colorectal Cancer
- ROLE OF PHOSPHATIDIC ACID IN CONTRACTILE MECHANISMS OF SMOOTH MUSCLE
- Comparative Study of Calcium Ion Dynamics and Contractile Response in Rat Middle Cerebral and Basilar Arteries
- Lysophosphatidic Acid (LPA) Induces Plasma Exudation and Histamine Release in Mice via LPA Receptors
- Involvement of integrins in mechanical stress-induced local Ca^ influx, Ca^ spots, in the presence of lysophosphatidic acid in cultured lens epithelial cells
- Optical Bioimaging : From Living Tissue to a Single Molecule : Calcium Imaging in Blood Vessel In Situ Employing Two-Photon Excitation Fluorescence Microscopy
- Lysophosphatidic Acid Enhances In Vivo Infiltration and Activation of Guinea Pig Eosinophils and Neutrophils via a Rho/Rho-Associated Protein Kinase-Mediated Pathway
- Physiological and Pharmacological Role of Lysophosphatidic Acid as Modulator in Mechanotransduction
- Changes in Ca^ Signaling and Contractile Protein Isoforms in Smooth Muscle Cells from Guinea Pig Ileum during Culture
- Contractile Protein Isoforms of Single and Cultured Smooth Muscle Cells from Guinea Pig Ileum
- CHANGES IN CONTRACTILE RESPONSES OF SINGLE SMOOTH MUSCLE CELLS TO AGONISTS
- Inositol 1,4,5-Trisphosphate-Induced Contraction in Skinned Single Smooth Muscle Cells from Guinea-Pig Taenia Coli
- Identification and Expression of Frizzled-3 Protein in Rat Frontal Cortex After Antidepressant and Electroconvulsive Treatment
- STUDIES ON ISOLATED SMOOTH MUSCLE CELLS.VIII.^Ca-EFFLUX FROM SINGLE SMOOTH MUSCLE CELLS ISOLATED FROM TAENIA COLI OF GUINEA PIG
- STUDIES ON ISOLATED SMOOTH MUSCLE CELLS. IV. ISOLATION AND ACETYLCHOLINE-CONTRACTION OF SINGLE SMOOTH MUSCLE CELLS FROM TAENIA COLI OF GUINEA-PIG
- STUDIES ON ISOLATED SMOOTH MUSCLE CELLS. VII. RESPONSE TO AGONISTS AND THE CONTRACTION VELOCITY OF TAENIA COLI OF GUINEA PIG AND THE SINGLE SMOOTH MUSCLE CELLS
- Two Types of Cation Channel Activated by Stimulation of Muscarinic Receptors in Guinea-Pig Urinary Bladder Smooth Muscle
- Squamous Cell Carcinoma of the Breast: Three Case Reports
- Expression of Transforming Growth Factor-β Superfamily Receptors in Developing Rat Eyes
- Purification and Characterization of a Major Collagenase from Streptomyces parvulus
- A case of calcifying odontogenic cyst with numerous calcifications: immunohistochemical analysis
- Role of Nrf2 and p62/ZIP in the neurite outgrowth by carnosic acid in PC12h cells
- Parasympathetic Vasodilator Fibers in Masseter Muscle
- Parasympathetic Vasodilator Fibers in the Orofacial Region
- Fractionation of Mature Eosinophils in GATA-Reporter Transgenic Mice
- ULTRASTRUCTURE OF SINGLE SMOOTH MUSCLE CELLS CONTRACTED BY CARBACHOL AND CALCIUM ION
- Structure and Regulation of the Chicken GATA-3 Gene
- Regulation of Meiosis in Fission Yeast
- BINDING PROPERTIES OF HYDROPHILIC MUSCARINIC ANTAGONIST [^3H] NMeQNB IN THE SINGLE SMOOTH MUSCLE CELLS FROM GUINEA-PIG ILEUM
- Comparison of Effects of Famotidine on Vagally and Field - Electrically Stimulated Acid Secretion in the Isolated Mouse Whole Stomach
- Pharmacological Features of Non-adrenergic Non-cholinergic (NANC) Relaxation Induced by Electrical Vagal Stimulation in Isolated Mouse Stomach
- STUDIES ON ISOLATED SMOOTH MUSCLE CELLS. V. CALCIUM AND CAFFEINE CONTRACTION OF SINGLE SMOOTH MUSCLE CELLS ISOLATED FROM TAENIA COLI OF GUINEA PIG
- Transcriptional regulation by GATA1 and GATA2 during erythropoiesis
- EFFECTS OF PLURONIC F-127 ON LOADING OF FURA 2/AM INTO SINGLE SMOOTH MUSCLE CELLS ISOLATED FROM GUINEA PIG TAENIA COLI
- Nrf2 regulates NGF mRNA induction by carnosic acid in T98G glioblastoma cells and normal human astrocytes
- Nrf2 inhibits hepatic iron accumulation and counteracts oxidative stress-induced liver injury in nutritional steatohepatitis
- Studies on metabolic fate of a new antiallergic agent, azelastine (4-(p-chlorobenzyl)-2-[N-methylperhydroazepinyl-(4)]-1-(2H)-phthalazinone hydrochloride).
- Spontaneous Oscillations of Cytoplasmic Free Calcium Ion Concentration in Cultured Smooth Muscle Cells from Guinea Pig Ileum.
- Heterogeneous contractile responses of single smooth muscle cells from guinea pig ileum.
- The Relationship between Spontaneous Calcium Oscillations and Cell Proliferation in Cultured Smooth Muscle Cells.
- Conserved Structure, Regulatory Elements, and Transcriptional Regulation from the GATA-1 Gene Testis Promoter.
- Deletion of Nrf2 leads to rapid progression of steatohepatitis in mice fed atherogenic plus high-fat diet