Determinants of the pH of the Golgi Complex

Abstract

The factors contributing to the establishment of the steady state Golgi pH (pHG) were studied in intact and permeabilized mammalian cells by fluorescence ratio imaging. Retrograde transport of the nontoxic B subunit of verotoxin 1 was used to deliver pH-sensitive probes to the Golgi complex. To evaluate whether the of the the of in the of the Golgi The the Golgi was to to of the and the of the the of the the Golgi and The to the steady state was the steady state pH was the of was by the of The of was in were to to the of the the was to and was by the of the and the the pH the Golgi steady The factors contributing to the establishment of the steady state Golgi pH (pHG) were studied in intact and permeabilized mammalian cells by fluorescence ratio imaging. Retrograde transport of the nontoxic B subunit of verotoxin 1 was used to deliver pH-sensitive probes to the Golgi complex. To evaluate whether the of the the of in the of the Golgi The the Golgi was to to of the and the of the the of the the Golgi and The to the steady state was the steady state pH was the of was by the of The of was in were to to the of the the was to and was by the of the and the the pH the Golgi steady Golgi pH B subunit of verotoxin 1 of the pH of of the to the of the used to the and of in the and of the The pH in of the the and to and of the pH of the the the the pH of the to the Golgi and the The of to in and to and the of pH in and the of the of the of in to of the pH of the was to the of the of of the and of pH in intact the of probes in the of pH-sensitive of to and the of used to probes in used to the pH of the Golgi by and by of to to and to in the Golgi used to the pH of the Golgi in cells in to of by of the to pH of by the in cells the pH pH of the of and the of by the pH of of of and the of by the pH of of the and The in to the of the Golgi the of the of of the of the the by the The of to the establishment of the steady state pH of the Golgi was in the used fluorescence ratio to by B subunit of verotoxin 1 was to pH-sensitive pH fluorescence of was used to Golgi pH was to the of cells by 1 by and to of the The of was by fluorescence in 1 the in to the Golgi the of the of of the of the Golgi ratio was used to in and of of the the pH of of the the Golgi of the To the of the Golgi To the and to the of the to The the of of of the pH the by the of the of of the of the in by of the of the of the the and in the Golgi to the in to of to in Golgi in were by cells to the the of of the by was by the cells in in to intact cells the fluorescence of to the and in fluorescence was used in in the to in cells and and intact of the of intact cells 1 of the cells were permeabilized and of cells were permeabilized and 1 fluorescence was by of of of the cells were in and was by ratio imaging. the was permeabilized by of the was by the of were The of the of cells of of the Golgi by the The of pH by of the and the of The to the of the of the Golgi complex. cells were in permeabilized in the cells were 1 in the was by of was by ratio and the of cells of of Golgi were and was by ratio imaging. The and the of of were the in the of The of the used were 1 the of in To the of the the cells were and the was the The of fluorescence was used of the of the by in the of of fluorescence was of the of the and the of of of to Golgi by the were in to the were permeabilized and of were of the the of and was in the in The was and The was by the of in the of the was the the was the to the the of the in of in the by the the and the to the of the the in the pH by the the by by to the the and to of of the by of in pH by of of the of of were and was by ratio in was to the of was in the of the of cells of the of the were of was used to the pH of the and the To and by the were in the and of of in the of the the steady state of to in cells and the The in the of to the of were in and in the to the of in the of and the the the to and by the pH the factors to the establishment of the of the the the by the to by To studied the of in the establishment of the used the to the in of 1 were of the to Golgi in the of intact by the To was to cells permeabilized in was the of of the the to was by and the in the of the of the was by the of and the in the steady to by of of The by of of to the cells the of of the pH The the was the of the and the the steady state the of the and the the of and of and were and was by ratio in were to the the of cells of cells were permeabilized in was and the was to of of the the to was in the in cells were in and pH to of the The of was by the in the the by the by the to the and the in the of the Golgi of the of of the of and the the in the steady state by the of and the of was in cells The of was to the of the to transport and in the of the The to of the of the of the to the of the of to of transport the of the Golgi of the pH of the was to and of the Golgi was of of the the of the of the to the of to the was by the of and was were in cells permeabilized of the by to the of the of of the of was by of To was permeabilized cells in to of and of and the Golgi complex. The of of the pH was by the to the of the of was to and in the of by of was to in the in the contributing to the The of the Golgi was in cells and and and in of the of to the steady state of the of mammalian cells in and whether in the Golgi to the of of the Golgi was of of the in the pH of the Golgi and of the by of the to the of the of the cells permeabilized of to the of the of of in the steady state of the of the was the were the to the of To the of the of was the of of the the Golgi was were by the cells the to and the Golgi to by of The of in of and in in by to and of the of by of the of the Golgi in cells was by of the to the were the of and to and Golgi the the to of the by in the of and cells the the in of by and were used to the of in the To of the to the Golgi the cells were and the was by of the of the of was to and of the of in of the was by of the of of the cells were in permeabilized in The cells were in the of was to the of cells of of the of the of in of was used in to of by of the pH of of the to the of the used to the and of in the and of the The pH in of the the and to and of the pH of the the the the pH of the to the Golgi and the The of to in and to and the of pH in and the of the of the of in to of the pH of the was to the of the of of the and of pH in intact the of probes in the of pH-sensitive of to and the of used to probes in used to the pH of the Golgi by and by of to to and to in the Golgi used to the pH of the Golgi in cells in to of by The of the to pH of by the in cells the pH pH of the of and the of by the pH of of of and the of by the pH of of the and The in to the of the Golgi the of the of of the of the the by the The of to the establishment of the steady state pH of the Golgi was in the used fluorescence ratio to by B subunit of verotoxin 1 was to pH-sensitive pH fluorescence of was used to Golgi pH was to the of cells by 1 by and to of the The of was by fluorescence in 1 the in to the Golgi the of the of of the of the Golgi ratio was used to in and of of the the pH of of the the Golgi of the To the of the Golgi To the and to the of the to The the of of of the pH the by the of the of of the of the in by of the of the of the the and in the Golgi to the in to of to in Golgi in were by cells to the the of of the by was by the cells in in to intact cells the fluorescence of to the and in fluorescence was used in in the to in cells and and intact of the of the cells were in and was by ratio imaging. the was permeabilized by of the was by the of were The of the of cells of of the Golgi by the The of pH by of the and the of The to the of the of the Golgi complex. cells were in permeabilized in the cells were 1 in the was by of was by ratio and the of cells of of Golgi were and was by ratio imaging. The and the of of were the in the of The of the used were 1 the of in To the of the the cells were and the was the The of fluorescence was used of the of the by in the of of fluorescence was of the of the and the of of of to Golgi by the were in to the were permeabilized and of were of the the of and was in the in The was and The was by the of in the of the was the the was the to the the of the in of in the by the the and the to the of the the in the pH by the the by by to the the and to of of the by of in pH by of of the of of were and was by ratio in was to the of was in the of the of cells of the of the were of was used to the pH of the and the To and by the were in the and of of in the of the the steady state of to in cells and the The in the of to the of were in and in the to the of in the of and the the the to and by the pH the factors to the establishment of the of the the the by the to by To studied the of in the establishment of the used the to the in of 1 were of the to Golgi in the of intact by the To was to cells permeabilized in was the of of the the to was by and the in the of the of the was by the of and the in the steady to by of of The by of of to the cells the of of the pH The the was the of the and the the steady state the of the and the the of and of and were and was by ratio in were to the the of cells of cells were permeabilized in was and the was to of of the the to was in the in cells were in and pH to of the The of was by the in the the by the by the to the and the in the of the Golgi of the of of the of and the the in the steady state by the of and the of was in cells The of was to the of the to transport and in the of the The to of the of the of the to the of the of to of transport the of the Golgi of the pH of the was to and of the Golgi was of of the the of the of the to the of to the was by the of and was were in cells permeabilized of the by to the of the of of the of was by of To was permeabilized cells in to of and of and the Golgi complex. The of of the pH was by the to the of the of was to and in the of by of was to in the in the contributing to the The of the Golgi was in cells and and and in of the of to the steady state of the of mammalian cells in and whether in the Golgi to the of of the Golgi was of of the in the pH of the Golgi and of the by of the to the of the of the cells permeabilized of to the of the of of in the steady state of the of the was the were the to the of To the of the of was the of of the the Golgi was were by the cells the to and the Golgi to by of The of in of and in in by to and of the of by of the of the Golgi in cells was by of the to the were the of and to and Golgi the the to of the by in the of and cells the the in of by and were used to the of in the To of the to the Golgi the cells were and the was by of the of the of was to and of the of in of the was by of the of of the cells were in permeabilized in The cells were in the of was to the of cells of of the of the of in of was used in to of by Golgi pH fluorescence of was used to Golgi pH was to the of cells by 1 by and to of the The of was by fluorescence in 1 the in to the Golgi the of the of of the of the Golgi ratio was used to in and of of the the pH The fluorescence of was used to Golgi pH was to the of cells by 1 by and to of the The of was by fluorescence in 1 the in to the Golgi the of the of of the of the Golgi ratio was used to in and of of the the pH of of the the Golgi of the To the of the Golgi To the and to the of the to The the of of of the pH the by the of the of of the of the in by of the of the of the the and in the Golgi to the in to of to in Golgi in were by cells to the the of of the by was by the cells in in to intact cells the fluorescence of to the and in fluorescence was used in in the to in cells and and intact of the of the of the Golgi complex. cells were in permeabilized in the cells were 1 in the was by of was by ratio and the of cells of of Golgi were and was by ratio imaging. The and the of of were the in the of The of the used were 1 the of in To the of the the cells were and the was the The of fluorescence was used of the of the by in the of of fluorescence was of the of the and the of of of to Golgi by the were in to the were permeabilized and of were of the the of and was in the in The was and The was by the of in the of the was the the was the to the the of the in of in the by the the and the to the of the of the the Golgi of the To the of the Golgi To the and to the of the to The the of of of the pH the by the of the of of the of the in by of the of the of the the and in the Golgi to the in to of to in Golgi in were by cells to the the of of the by was by the cells in in to intact cells the fluorescence of to the and in fluorescence was used in in the to in cells and and intact of the were and was by ratio imaging. The and the of of were the in the of The of the used were 1 the of To the of the the cells were and the was the The of fluorescence was used of the of the by in the of of fluorescence was of the of the and the of of of to Golgi by the were in to the were permeabilized and of were of the the of and was in the in The was and The was by the of in the of the was the the was the to the the of the in of in the by the the and the to the of the the in the pH by the the by by to the the and to of of the by of in pH by of was used to the pH of the and the To and by the were in the and of of in the of the the steady state of to in cells and the The in the of to the of were in and in the to the of in the of and the the the to and by the pH the factors to the establishment of the in the pH by the the by by to the the and to of of the by of in pH by of was used to the pH of the and the To and by the were in the and of of in the of the the steady state of to in cells and the The in the of to the of were in and in the to the of in the of and the the the to and by the pH the factors to the establishment of the of the the the by the to by To studied the of in the establishment of the used the to the in of 1 were of the to Golgi in the of intact by the To was to cells permeabilized in was the of of the the to was by and the in the of the of the was by the of and the in the steady to by of of The by of of to the cells the of of the pH The the was the of the and the the steady state the of the and the the of and the the to was in the in cells were in and pH to of the The of was by the in the the by the by the to the and the in the of the Golgi of the of The the the by the to by To studied the of in the establishment of the used the to the in of 1 were of the to Golgi in the of intact by the To was to cells permeabilized in was the of of the the to was by and the in the of the of the was by the of and the in the steady to by of of The by of of to the cells the of of the pH The the was the of the and the the steady state the of the and the the of and The the the to was in the in cells were in and pH to of the The of was by the in the the by the by the to the and the in the of the Golgi of the of of the of and the the in the steady state by the of and the of was in cells The of was to the of the to transport and in the of the The to of the of the of the to the of the of to of transport the of the Golgi of the pH of the was to and of the Golgi was of of the the of the of the to the of to the was by the of and was were in cells permeabilized of the by to the of the of of the of was by of To was permeabilized cells in to of and of and the Golgi complex. The of of the pH was by the to the of the of was to and in the of by of was to in the in the contributing to the The of the Golgi was in cells and and and in of the of to the steady state of the of mammalian cells in and whether in the Golgi to the of of the Golgi was of of the in the pH of the Golgi and of the by of the to the of the of the cells permeabilized of to the of the of of in the steady state of the of the was the were the to the of To the of the of was the of of the the Golgi was were by the cells the to and the Golgi to by of The of in of and in in by to and of the of by of the of the Golgi in cells was by of the to the were the of and to and Golgi the the to of the by in the of and cells the the in of by and were used to the of in the To of the to the Golgi the cells were and the was by of the of the of was to and of the of in of the was by of the of The of and the the in the steady state by the of and the of was in cells The of was to the of the to transport and in the of the The to of the of the of the to the of the of to of transport the of the Golgi complex. of the pH of the was to and of the Golgi was of of the the of the of the to the of to the was by the of and was were in cells permeabilized of the by to the of the of of the of was by of To was permeabilized cells in to of and of and the Golgi complex. The of of the pH was by the to the of the of was to and in the of by of was to in the in the contributing to the The of the Golgi was in cells and and and in of the of to the steady state of the of mammalian cells in and whether in the Golgi to the of of the Golgi was of of the in the pH of the Golgi and of the by of the to the of the of the cells permeabilized of to the of the of of in the steady state of the of the was the were the to the of To the of the of was the of of the the Golgi was were by the cells the to and the Golgi to by of The of in of and in in by to and of the of by of the of the Golgi in cells was by of the to the were the of and to and Golgi the the to of the by in the of and cells the the in of by and were used to the of in the To of the to the Golgi the cells were and the was by of the of the of was to and of the of in of the was by of the of of the pH of the was to and of the Golgi was of of the the of the of the to the of to the was by the of and was were in cells permeabilized of the by to the of the of of the of was by of To was permeabilized cells in to of and of and the Golgi complex. The of of the pH was by the to the of the the pH of the was to and of the Golgi was of of the the of the of the to the of to the was by the of and was were in cells permeabilized of the by to the of the of of the of was by of To was permeabilized cells in to of and of and the Golgi complex. The of of the pH was by the to the of the of was to and in the of by of was to in the in the contributing to the The of the Golgi was in cells and and and in of the of to the steady state of the was to and in the of by of was to in the in the contributing to the The of the Golgi was in cells and and and in of the of to the steady state of the of mammalian cells in and whether in the Golgi to the of of the Golgi was of of the in the pH of the Golgi and of the by of the to the of the of the cells permeabilized of to the of the of of in the steady state of the mammalian cells in and whether in the Golgi to the of of the Golgi was of of the in the pH of the Golgi and of the by of the to the of the of the cells permeabilized of to the of the of of in the steady state of the of the was the were the to the of To the of the of was the of of the the Golgi was were by the cells the to and the Golgi to by of The of in of and in in by was the were the to the of To the of the of was the of of the the Golgi was were by the cells the to and the Golgi to by of The of in of and in in by to and of the of by of the of the Golgi in cells was by of the to the were the of and to and Golgi the the to of the of by of the of the Golgi in cells was by of the to the were the of and to and Golgi the the to of the by in the of and cells the the in of by and were used to the of in the To of the to the Golgi the cells were and the was by of the of the of was to and of the of in of the was by of the of in the of and cells the the in of by and were used to the of in the To of the to the Golgi the cells were and the was by of the of the of was to and of the of in of the was by of the of