Rewarming Of Cells Research Articles

Lumenal targeted GFP, used as a marker of soluble cargo, visualises rapid ERGIC to Golgi traffic by a tubulo-vesicular network.

The mechanism by which soluble proteins without sorting motifs are transported to the cell surface is not clear. Here we show that soluble green fluorescent protein (GFP) targeted to the lumen of the endoplasmic reticulum but lacking any known retrieval, retention or targeting motifs, was accumulated in the lumen of the ERGIC if cells were kept at reduced temperature. Upon activation of anterograde transport by rewarming of cells, lumenal GFP stained a microtubule-dependent, pre-Golgi tubulo-vesicular network that served as transport structure between peripheral ERGIC-elements and the perinuclear Golgi complex. Individual examples of these tubular elements up to 20 microm in length were observed. Time lapse imaging indicated rapid anterograde flow of soluble lumenal GFP through this network. Transport tubules, stained by lumenal GFP, segregated rapidly from COPI-positive membranes after transport activation. A transmembrane cargo marker, the temperature sensitive glycoprotein of the vesicular stomatitis virus, ts-045 G, is also not present in tubules which contained the soluble cargo marker lum-GFP. These results suggest a role for pre-Golgi vesicular tubular membranes in long distance anterograde transport of soluble cargo. http://www. biologists.com/JCS/movies/jcs1334.html

Active transport of potassium and chloride in an identifiable molluscan neuron.

Direct measurements of intracellular K(+) and Cl(-)activities before and after blockage of cellular metabolic processes indicate that K(+) is actively transported inwardly and Cl(-)is actively transported outwardly from the giant cell of the abdominal ganglion of Aplysia. The rewarming of cells that have been cooled to 1 degrees +/- 1 degrees C causes K(+) to be taken up and Cl(-) to be extruded against electrochemical gradients.