Compound Exocytosis in Pituitary Cells (original) (raw)
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Subnanometer Fusion Pores in Spontaneous Exocytosis of Peptidergic Vesicles
Journal of Neuroscience, 2007
Kiss-and-run exocytosis, consisting of reversible fusion between the vesicle membrane and the plasma membrane, is considered to lead to full fusion after stimulation of vesicles containing classical transmitters. However, whether this is also the case in the fusion of peptidergic vesicles is unknown.
‘Full fusion’ is not ineluctable during vesicular exocytosis of neurotransmitters by endocrine cells
Proceedings of the Royal Society A: Mathematical, Physical and Engineering Sciences, 2017
Vesicular exocytosis is an essential and ubiquitous process in neurons and endocrine cells by which neurotransmitters are released in synaptic clefts or extracellular fluids. It involves the fusion of a vesicle loaded with chemical messengers with the cell membrane through a nanometric fusion pore. In endocrine cells, unless it closes after some flickering (‘Kiss-and-Run’ events), this initial pore is supposed to expand exponentially, leading to a full integration of the vesicle membrane into the cell membrane—a stage called ‘full fusion’. We report here a compact analytical formulation that allows precise measurements of the fusion pore expansion extent and rate to be extracted from individual amperometric spike time courses. These data definitively establish that, during release of catecholamines, fusion pores enlarge at most to approximately one-fifth of the radius of their parent vesicle, hence ruling out the ineluctability of ‘full fusion’.
Cytosolic calcium facilitates release of secretory products after exocytotic vesicle fusion
FEBS Letters, 1995
We monitored single vesicle exocytosis by simultaneous measurements of cell membrane capacitance as an indicator of fusion and amperometric detection of serotonin release. We show here that vesicle-plasma membrane fusion in rat mast cell granules is followed by a variable, exponentially distributed, delay before bulk release. This delay reflects the time required for the expansion of the exocytotic fusion pore, lasting, on average, 231 ms in resting cytosolic calcium, [Ca2+L (50 nM). In the presence of [Ca2+]i in the low micromollar range, the lag between fusion and release was reduced to 123 ms. The characteristics of the amperometric signals were unchanged by [Ca2+]i. These resuits show a novel site of regulation in the exocytotic process, the fusion pore, which may represent a different mechanism facilitating transmitter release.
ChemPhysChem, 2003
Vesicular exocytosis is important in the communication between cells in complex organisms. It controls the release of specific chemical or biochemical messengers stored in the emitting cell, which elicit a response upon detection by the target cells. Secretion of a messenger molecule (a neurotransmitter) was measured electrochemically, which allowed the quantification of cellular events and the validation of current physicochemical models. This model led us to formulate predictions about the occurrence and kinetics of vesicular exocytotic events based on the physicochemical meaning of its key parameters. These predictions were tested successfully through a series of experiments on chromaffin cells, involving changes of osmotic conditions, presence of trivalent ions and cholesterol-induced structuring of the cell plasmic membrane.
Elementary properties of spontaneous fusion of peptidergic vesicles: fusion pore gating
The Journal of Physiology, 2007
The release of hormones and neurotransmitters by regulated exocytosis requires the delivery of secretory vesicles to the plasma membrane, where they dock and become primed for fusion with the plasma membrane. Upon stimulation a fusion pore is formed through which cargo molecules diffuse out of the vesicle lumen into the extracellular space. After the cargo release the fusion pore either closes (kiss-and-run, transient exocytosis), fluctuates between an open and a closed state (for short times, fusion pore flickering, or for rather longer periods, 'pulsing pore') or expands irreversibly (full fusion exocytosis). In almost all secretory cells spontaneous secretion of vesicle cargo can be detected in the absence of stimulation. Spontaneous and stimulated exocytosis were thought to exhibit similar properties at elementary level, differing only in the probability of occurrence. However, recent studies indicate that spontaneous exocytosis differs from the stimulated one in many respects, therefore opening questions about the physiological role of spontaneous exocytosis. In this report we address the elementary properties of spontaneous and stimulated peptidergic vesicle discharge which appears to be modulated by fusion pore conductance (diameter) and fusion pore gating.
Journal of neurochemistry, 2016
The pattern of stimulation defines important characteristics of the secretory process in neurons and neuroendocrine cells, including the pool of secretory vesicles being recruited, the type and amount of transmitters released, the mode of membrane retrieval and the mechanisms associated with vesicle replenishment. This review analyzes the mechanisms that regulate these processes in chromaffin cells, as well as in other neuroendocrine and neuronal models. A common factor in these mechanisms is the spatial and temporal distribution of the Ca(2+) signal generated during cell stimulation. For instance, neurosecretory cells and neurons have pools of vesicles with different locations with respect to Ca(2+) channels, and those pools are therefore differentially recruited following different patterns of stimulation. In this regard, a brief stimulus will induce the exocytosis of a small pool of vesicles that is highly coupled to voltage-dependent Ca(2+) channels, whereas longer or more inten...
The Fusion Pore and Vesicle Cargo Discharge Modulation
Annals of the New York Academy of Sciences, 2009
Exocytosis, the merger of the vesicle membrane with the plasma membrane, is thought to mediate the release of hormones and neurotransmitters from secretory vesicles. The work of Bernard Katz and colleagues decades ago considered that vesicle cargo discharge initially requires the delivery of secretory vesicles to the plasma membrane where vesicles dock and are primed for fusion with the plasma membrane. Then, upon stimulation, the vesicle and the plasma membranes fuse to form a transient fusion pore through which cargo molecules diffuse out of the vesicle lumen into the extracellular space. Katz and colleagues considered this process to occur in an all-or-none fashion. However, recent studies show that this may not be so simple. The aim of this overview is to highlight the novel findings that indicate that fusion pores are subject to regulations, which affect the release competence of a single vesicle. Here we discuss the elementary properties of spontaneous and stimulated peptidergic vesicle discharge, which appears to be modulated, at least in pituitary lactotrophs, by fusion pore conductance (pore diameter) and fusion pore gating (kinetics).
Vesicle-membrane fusion. Observation of simultaneous membrane incorporation and content release
Biophysical Journal, 1988
Vesicle fusion, the central process of neurotransmitter release and hormonal secretion, is a complex process culminating in simultaneous incorporation of vesicle membrane into the plasma membrane and release of the vesicular contents extracellularly. This report describes simulteous observation of membrane incorporation and content release using a model system composed of a planar bilayer and dye-filled vesicles.
Temporally resolved, independent stages of individual exocytotic secretion events
Biophysical journal, 1996
The stages of the complex events involved in exocytotic secretion after vesicle-cell membrane fusion have been examined at the level of individual vesicles. Catecholamine flux from single bovine adrenal medullary cells was measured with carbon-fiber microelectrodes firmly touching the cell surface. The data reveal that secretion during exocytotic events has three distinct stages: a small increase in catecholamine flux, a rapid, but not instantaneous, rise to a maximum, followed by an exponential decrease in the flux. These stages are interpreted in the following ways. The initial stage corresponds to catecholamine secretion through a fusion pore. The rate of pore expansion appears to control the rise time of the flux to its maximum value. The final exponential stage is consistent with chemical dissociation of the intravesicular matrix or gel.