The exocytotic fusion pore of small granules has a conductance similar to an ion channel.
We measured capacitance changes in cell attached patches of human neutrophils using a high frequency lock-in method. With this technique the noise level is reduced to 0.025 fF such that capacitance steps of 0.1 fF are clearly detected corresponding to exo- and endocytosis of single 60 nm vesicles. It is thus possible to detect almost all known exocytotic and endocytotic processes including exocytosis of small neurotransmitter containing vesicles in most cell types as well as endocytosis of coated and uncoated pits. In neutrophils we demonstrate a stepwise capacitance decrease generated by 60-165 nm vesicles as expected for endocytosis of coated and non-coated pits. Following ionomycin stimulation a stepwise capacitance increase is observed consisting of 0.1-5 fF steps corresponding to the different granule types of human neutrophils from secretory vesicles to azurophil granules. The opening of individual fusion pores is resolved during exocytosis of 200 nm vesicles. The initial conductance has a mean value of 150 pS and can be as low as 35 pS which is similar to the conductance of many ion channels suggesting that the initial fusion pore is formed by a protein complex.
Lollike
K
K
Department of Internal Medicine and Haematology L, National University Hospital, Copenhagen, Denmark.
Borregaard
N
N
Lindau
M
M
D003594:Cytoplasmic Granules / Q000502:physiology*; D004553:Electric Conductivity; D004705:Endocytosis; D005089:Exocytosis / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D007473:Ion Channels / Q000502:physiology*; D015759:Ionomycin / Q000494:pharmacology; D008561:Membrane Fusion; D008564:Membrane Potentials; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques
D007473:Ion Channels; D015759:Ionomycin