Electron-probe analysis of isolated goldfish hair cells: implications for preparing healthy cells.
Electron-probe analysis provides an objective criterion for the physiological status of cells: whether they show the high potassium and low sodium that are expected of healthy animal cells. Preparing isolated goldfish hair cells that were healthy by this criterion required several precautions, including: limited exposure to enzymes and to simple salt solutions, a rest period between enzyme treatment and mechanical disruption of the tissue, and presence of bovine albumin in the medium both during the rest period and during mechanical dispersion and plating. Cells prepared with these precautions from the saccule and lagena and kept in an enriched medium had the following elemental composition (mole percentages with respect to phosphorus): K, 103; Na, 18; Cl, 23; S, 13; Mg, 8; Ca, 1.5. These mole percentages were close to these elements' total millimolar concentrations in the cells. If the precautions were not taken, cells with intact surface membranes (as assessed by exclusion and retention of dyes) could be obtained, but the cells had elevated cell sodium and low cell potassium.
Mroz
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Eaton-Peabody Laboratory, Massachusetts Eye and Ear Infirmary, Boston 02114.
Nissim
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KR
Lechene
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D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002712:Chlorides / Q000378:metabolism; D003470:Culture Media; D004577:Electron Probe Microanalysis; D004602:Elements; D005069:Evaluation Studies as Topic; D006054:Goldfish / Q000378:metabolism*; D006198:Hair Cells, Auditory / Q000737:chemistry* / Q000166:cytology* / Q000378:metabolism; D066298:In Vitro Techniques; D007473:Ion Channels / Q000378:metabolism; D017136:Ion Transport; D011188:Potassium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism
D002712:Chlorides; D003470:Culture Media; D004602:Elements; D007473:Ion Channels; D012964:Sodium; D011188:Potassium