title: Membrane-labeled MDCK cells and confocal microscopy for the analyses of cellular volume and morphology.
abstract: A clone of Madin-Darby canine kidney (MDCK) cells whose cell membrane was stably labeled with expressed cyan fluorescent protein (CFP) was established, and changes in their volume and shape induced by hyposmotic stress were analyzed with confocal microscopy. The membrane-targeted CFP was present not only on the cell membrane but also in the endoplasmic reticulum and Golgi apparatus, but was excluded from the mitochondria and cell nucleus. During hyposmosis, the initial swelling and the following regulatory volume decrease could be accurately measured by summation of the cellular volume in every confocal slice crossing the cell at different heights. Changes in the cellular height roughly paralleled the changes in cellular volume when the mean value was compared, but dissociation as much as 30% was observed for individual cells due to changes in cell shape. The present experimental system, which enables direct measurement of cell volume and simultaneous observation of various intracellular phenomena, would be useful for further investigation of cellular volume regulation.
authors: Department of Food Science, Otsuma Women's University, Chiyoda-ku, Tokyo 102-8357, Japan.
authors: Naoko
authors: N
authors: Iida-Tanaka
authors: Iyuki
authors: I
authors: Namekata
authors: Miku
authors: M
authors: Tamura
authors: Yuko
authors: Y
authors: Kawamata
authors: Toru
authors: T
authors: Kawanishi
authors: Hikaru
authors: H
authors: Tanaka
mesh_terms: D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D048429:Cell Size; D002477:Cells / Q000648:ultrastructure*; D002999:Clone Cells; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000648:ultrastructure; D049452:Green Fluorescent Proteins; D018613:Microscopy, Confocal / Q000379:methods*; D009997:Osmotic Pressure; D010169:Palmitic Acids / Q000378:metabolism
chemical_list: C485182:Cyan Fluorescent Protein; D010169:Palmitic Acids; D049452:Green Fluorescent Proteins