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Journal of Neuroscience, Vol 16, 553-562, Copyright © 1996 by Society for Neuroscience
Kinetics of stimulus-coupled secretion in dialyzed bovine chromaffin cells in response to trains of depolarizing pulses
EP Seward and MC Nowycky
Department of Anatomy and Neurobiology, Medical College of Pennsylvania, Philadelphia 19129, USA.
Stimulus-secretion coupling in bovine chromaffin cells was investigated
with whole-cell patch-clamp recordings and capacitance detection techniques
to monitor exocytosis in response to trains of depolarizing pulses. Two
kinetically discrete modes of exocytotic responses were observed. In one
mode, the first depolarization of a train elicited a large increase in
membrane capacitance (Cm; mean approximately 70 fF). This secretory mode
was characterized by small Ca2+ requirements, relative insensitivity to the
pipette Ca2+ chelator concentration, and rapid depletion of the secretory
response. This mode of stimulus- secretion coupling was labile and was seen
only in response to the first and, occasionally, the second stimulus train
of whole-cell recordings. The second type of exocytotic response persisted
for the remainder of the whole-cell recordings and consisted of two
distinct phases. During the earliest pulses of a stimulus train, Ca2+ entry
did not evoke Cm increases. Instead, Cm responses were elicited by later
pulses, despite diminished Ca2+ entry per pulse caused by Ca2+ channel
inactivation. The secretory phase was initiated after a specific
"threshold" amount of Ca2+ had entered the cell, which was determined by
the concentration, but not the binding kinetics, of the Ca2+ chelator in
the pipette. In both the early and the secretory phases, the response of
the cell was proportional to cumulative Ca2+ entry, regardless of current
amplitude, pulse duration, or number of pulses. Threshold-type secretory
kinetics has been described previously in peptide-secreting
neurohypophysial (NHP) nerve terminals (Seward et al., 1995). Secretory
kinetics with minimal Ca2+ requirements has not been observed in that
preparation. Chromaffin cells appear to possess a broader repertoire of
stimulus-secretion coupling modes than NHP terminals.
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