Electropermeabilization of endocytotic vesicles in B16 F1 mouse melanoma cells

It has been reported previously that electric pulses of sufficiently high voltage and short duration can permeabilize the membranes of various organelles inside living cells. In this article, we describe electropermeabilization of endocytotic vesicles in B16 F1 mouse melanoma cells. The cells were exposed to short, high-voltage electric pulses (from 1 to 20 pulses, 60 ns, 50 kV/cm, repetition frequency 1 kHz). We observed that 10 and 20 such pulses induced permeabilization of membranes of endocytotic vesicles, detected by release of lucifer yellow from the vesicles into the cytosol. Simultaneously, we detected uptake of propidium iodide through plasma membrane in the same cells. With higher number of pulses permeabilization of the membranes of endocytotic vesicles by pulses of given parameters is accompanied by permeabilization of plasma membrane. However, with lower number of pulses only permeabilization of the plasma membrane was detected

Responses of plant cells and tissues to pulsed electric field treatments

Cell membrane electroporation/permeabilization may be achieved without affecting cell viability through strict control of the electric pulse parameters. This process is referred to as reversible permeabilization. Even if the cells survive the electric field treatment, they are subjected to stress due to the opening of pores and the struggle of the cells to recover their normal functionality. Very little is known about what actually occurs in the cell and its membranes at the molecular level upon reversible electroporation, and the physiological responses to pulsed electric field (PEF)-induced stress are still largely unknown. This chapter explores the current state of the art on the influence of the complexity of plant tissues on electroporation. Focusing on reversible electroporation, metabolic responses of plant cells and tissues induced by PEF application are also reviewed. One of the first challenges when electroporating plant tissue is their heterogeneous structures where cells vary in shape, size, and cell wall structure. This heterogeneity influences the effect of different electric fields protocols aiming at permeabilizing all cells in the tissue. Once cells are reversibly permeabilized, physiological responses to PEF-induced stress include the production of reactive oxygen species, mobilization of stored energy, activation of stress-related genes, and the production of secondary metabolites. The application of reversible PEF has also been shown to barley seed germination as well as to increase the strength of the cell wall in potatoes and, in consequence, their textural properties. This chapter finishes by revising the effect of reversible PEF on protoplasts (plant cells where the cell walls have been removed) and, in consequence, on the regeneration of new plants. Overall, reports on reversible permeabilization of plant cells and tissues are not common in the literature; however, they have laid the foundation for a fascinating area of research and technological innovation