Patch-clamp techniques for time-resolved capacitance measurements in single cells

1988 ◽  
Vol 411 (2) ◽  
pp. 137-146 ◽  
Author(s):  
Manfred Lindau ◽  
Erwin Neher
Keyword(s):  
Patch Clamp ◽  
Single Cells ◽  
Time Resolved ◽  
Capacitance Measurements

Time–resolved capacitance measurements: monitoring exocytosis in single cells

1991 ◽  
Vol 24 (1) ◽  
pp. 75-101 ◽  
Author(s):  
Manfred Lindau
Keyword(s):  
Plasma Membrane ◽  
Mast Cells ◽  
Endocrine Cells ◽  
Secretory Granules ◽  
Single Cells ◽  
Time Resolved ◽  
Capacitance Measurements ◽  
Fusion Pores

Many cells release preformed material contained in secretory granules by exocytosis. Exocytosis is a specialized means of secretion in which the granules fuse with the plasma membrane and thereby discharge their contents through the fusion pores. This mechanism mediates, for example, the formation of the fertilization envelope in eggs, the release of neurotransmitters and neuropeptides by neurons, the release of a variety of enzymes and mediators by mast cells and granulocytes or the secretion of hormones by endocrine cells. Classical methods for investigating exocytosis usually measure release of secreted material.

Patch-Clamp Recording and RT-PCR on Single Cells

2003 ◽  
pp. 193-232 ◽  
Author(s):  
Bertrand Lambolez ◽  
Etienne Audinat ◽  
Pascal Bochet ◽  
Jean Rossier
Keyword(s):  
Patch Clamp ◽  
Single Cells ◽  
Rt Pcr ◽  
Patch Clamp Recording

scholarly journals Regulation of transferrin-induced endocytosis by wild-type and C282Y-mutant HFE in transfected HeLa cells

2002 ◽  
Vol 282 (5) ◽  
pp. C973-C979 ◽  
Author(s):  
Lukas Schwake ◽  
Andreas W. Henkel ◽  
Hans D. Riedel ◽  
Thorsten Schlenker ◽  
Matthias Both ◽  
...  
Keyword(s):  
Hela Cells ◽  
Single Cells ◽  
Hereditary Hemochromatosis ◽  
Inhibitory Effect ◽  
Membrane Capacitance ◽  
Wild Type ◽  
Time Resolved ◽  
Cell Membrane Capacitance ◽  
Endocytic Vesicles ◽  
In Cells

The hereditary hemochromatosis protein HFE is known to complex with the transferrin receptor; however, its function regarding endocytosis of transferrin is unclear. We performed patch-clamp capacitance measurements in transfected HeLa cells carrying wild-type or C282Y-mutant HFE cDNA under the control of a tetracycline-sensitive promoter. Whole cell experiments in cells with suppressed expression of wild-type HFE revealed a decrease in membrane capacitance, reflecting predominance of endocytosis in the presence of transferrin. Cells overexpressing C282Y-mutant HFE displayed less intense capacitance decreases, whereas no significant decrease was observed in cells overexpressing wild-type HFE. The formation of single endocytic vesicles in cells with suppressed expression of wild-type HFE was greatly increased in the presence of transferrin as revealed by cell-attached recordings. According to their calculated diameters, many of these vesicles corresponded to clathrin-coated vesicles. These results suggest that wild-type HFE negatively modulates the endocytic uptake of transferrin. This inhibitory effect is attenuated in cells expressing C282Y-mutant HFE. Time-resolved measurements of cell membrane capacitance provide a powerful tool to study transferrin-induced endocytosis in single cells.

scholarly journals Massively parallel and time-resolved RNA sequencing in single cells with scNT-seq

2020 ◽  
Vol 17 (10) ◽  
pp. 991-1001 ◽  
Author(s):  
Qi Qiu ◽  
Peng Hu ◽  
Xiaojie Qiu ◽  
Kiya W. Govek ◽  
Pablo G. Cámara ◽  
...  
Keyword(s):  
Rna Sequencing ◽  
Single Cells ◽  
Massively Parallel ◽  
Time Resolved

Capacitance Measurements in the Mouse Rod Bipolar Cell Identify a Pool of Releasable Synaptic Vesicles

2006 ◽  
Vol 96 (5) ◽  
pp. 2539-2548 ◽  
Author(s):  
Zhen-Yu Zhou ◽  
Qun-Fang Wan ◽  
Pratima Thakur ◽  
Ruth Heidelberger
Keyword(s):  
Bipolar Cell ◽  
Synaptic Vesicles ◽  
Sine Wave ◽  
Membrane Capacitance ◽  
Membrane Properties ◽  
Intact Mouse ◽  
Time Resolved ◽  
Capacitance Measurements ◽  
Treatment Conditions ◽  
Capacitance Response

The mouse is an important model system for understanding the molecular basis of neuronal signaling and diseases of synaptic communication. However, the best-characterized retinal ribbon-style synapses are those of nonmammalian vertebrates. To remedy this situation, we asked whether it would be feasible to track synaptic vesicle dynamics in the isolated mouse rod bipolar cell using time-resolved capacitance measurements. The results demonstrate that membrane depolarization triggered an increase in membrane capacitance that was Ca2+ dependent and restricted to the synaptic compartment, consistent with exocytosis. The amplitude of the capacitance response recorded from the easily accessible soma of an intact mouse rod bipolar cell was identical to that recorded directly from the small synaptic terminal, suggesting that in the carefully selected cohort of cells presented here, axonal resistance was not a significant barrier to current flow. This supposition was supported by the analysis of passive membrane properties and a comparison of membrane capacitance measurements in cells with and without synaptic terminals and reinforced by the lack of an effect of sine-wave frequency (200–1,600 Hz) on the measured capacitance increase. The magnitude of the capacitance response increased with Ca2+ entry until a plateau was reached at a spatially averaged intraterminal calcium of about 600 nM. We interpret this plateau, nominally 30 fF, as corresponding to a releasable pool of synaptic vesicles. The robustness of this measure suggests that capacitance measurements may be used in the mouse rod bipolar cell to compare pool size across treatment conditions.

Development of a streak-camera-based time-resolved microscope fluorometer and its application to studies of membrane fusion in single cells

Biochemistry ◽  
1991 ◽  
Vol 30 (26) ◽  
pp. 6517-6527 ◽  
Author(s):  
Akihiro Kusumi ◽  
Akihiko Tsuji ◽  
Masayuki Murata ◽  
Yasushi Sako ◽  
Akiyasu C. Yoshizawa ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Streak Camera ◽  
Single Cells ◽  
Time Resolved

Impedance spectra of patch clamp scenarios for single cells immobilized on a lab-on-a-chip

2013 ◽  
Vol 17 (2) ◽  
pp. 263-274 ◽  
Author(s):  
M. Alberti ◽  
D. Snakenborg ◽  
J. M. Lopacinska ◽  
M. Dufva ◽  
J. P. Kutter
Keyword(s):  
Patch Clamp ◽  
Single Cells ◽  
Lab On A Chip ◽  
Impedance Spectra
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