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  • 1
    Electronic Resource
    Electronic Resource
    Laser microsurgery: a versatile tool in plant (electro) physiology (1994)
    Springer
    Protoplasma 178 (1994), S. 1-10 
    Details
    ISSN: 1615-6102
    Keywords: Elodea densa ; Eremosphaera ; Laser microsurgery ; Lilium longiflorum ; Patch-clamp ; Nicotiana tabacum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In plant cells the cell wall is a formidable obstacle in many physiological studies such as patch-clamp measurements and cell labelling with antibodies. Enzymatic digestion of the cell wall, in order to release a protoplast, has a number of disadvantages; therefore we worked out an alternative method to gain access to the plasma membrane. The wall of specialized cells from three higher plant species and one unicellular alga were perforated using the focussed UV light of a nitrogen laser. In order to enhance the absorption of the UV light by the walls, a dye was used that binds specifically to cell wall components. Extrusion of the protoplast or parts thereof was controlled by a regulated gradual decrease of the osmolarity of the solution surrounding the cells. Cytoplasmic streaming and chloroplast circulation were maintained in the protoplasts, demonstrating their viability after the wall perforation with the laser. Continuous deposition of new cell wall material by the polar tip of pollen tubes after surgical removal of the wall at the tip is another demonstration of the viability of the cells. Formation of high resistance seals between the plasma membrane and a patch pipet was surprisingly difficult. The role of ‘Hechtian strands’ and continuing synthesis of cell wall material in seal formation is further investigated. Other applications for the surgical laser are: fusion of two cells or vacuoles, analysis of the composition of specific parts of the cell wall, and release of the vacuole from an identified cell type for patchclamp studies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01404115
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  • 2
    Electronic Resource
    Electronic Resource
    Vacuolar ion currents in the primitive green algaEremosphaera viridis: The electrical properties are suggestive of both the Characeae and higher plants (1994)
    Springer
    Protoplasma 179 (1994), S. 34-45 
    Details
    ISSN: 1615-6102
    Keywords: Eremosphaera viridis ; Instantaneous current ; Time activated current ; Patch-clamp ; Phylogeny ; Vacuolar membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The electrical properties of the vacuolar membrane of the primitive green algaEremosphaera virdis were investigated using the patch-clamp technique. In whole vacuole measurements two types of transport systems with long activation time-constants were identified. The first, showing marked outward rectification, was activated by an increase in the cytosolic calcium concentration. Furthermore, it displayed sensitivity to micromolar concentrations of the anion channel blocker Zn2+ and to acidification of the cytosol. In contrast, the second time-activated current component was almost insensitive to changes in cytosolic pH and was blocked by the potassium channel inhibitor TEA. In addition to these slowly activating current components, the vacuolar membrane contained at least two further transport systems, responsible for an instantaneous current. These two current components were distinguished by their different sensitivity to protons, cytosolic calcium, and TEA. Comparing these electrical properties to those observed in vacuoles of higher plants or in cytoplasmic droplets from characean algae, respectively, it seems thatEremosphaera is intermediate, corresponding to the systematic position of this simple green alga.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01360735
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