Function Suggests Nano-Structure: Quantitative Structural Support for SNARE-Mediated Pore Formation.

Granule secretory content is released in either basal or calcium-activated complete exocytosis mode. A vital element in these processes is the establishment of a fusion pore between the granule membrane and the plasma membrane, initiated by the formation of a circular rosette docking arrangement of SNARE protein complexes. The controversially disputed number of SNARE complexes needed for granule priming leading to the formation of the fusion pore, is granule-size dependent and varies between secretion modes. Resorting to a statistical mechanics approach that views SNARE complexes and Ca(2+) ions as interacting particles, we have developed a relationship that links secretion rate to SNARE rosette size, Ca(2+) concentration and Ca(2+) ion cooperativity. Data are presented and discussed which suggest this SNARE-dependent generalization of existing narrow-range biophysical models that correlate secretion rate with Ca(2+) concentration and maximal Ca(2+) ion cooperativity. Evidence from dozens of examples in the literature advocate for this relation, which holds through the entire biological range. The coalescence of so many areas of diverse research methodologies has greatly augmented our understanding of so many different sequences of granule life cycle. Accordingly, these new tools may become valuable in a variety of electrophysiological experiments.

The role of synaptobrevin1/VAMP1 in Ca2+-triggered neurotransmitter release at the mouse neuromuscular junction.

Synaptobrevin (Syb)/vesicle-associated membrane protein (VAMP) is a small, integral membrane protein of synaptic vesicles. Two homologous isoforms of synaptobrevin, Syb1/VAMP1 and Syb2/VAMP2, exhibit distinct but partially overlapping patterns of expression in adult mammalian neurons: Syb1 is predominantly expressed in the spinal cord, especially in motor neurons and motor nerve terminals of the neuromuscular junction (NMJ), whereas Syb2 is primarily expressed in central synapses in the brain. Whereas many studies have focused on the function of Syb2 in the brain, few studies have examined the role of Syb1. Here we report that Syb1 plays a critical role in neuromuscular synaptic transmission. A null mutation of Syb1 resulting from a spontaneous, nonsense mutation in mice significantly impairs the function, but not the structure, of the NMJ. In particular, both spontaneous and evoked synaptic activities in Syb1 mutant mice are reduced significantly relative to control mice. Short-term synaptic plasticity in Syb1-deficient NMJs is markedly altered: paired-pulse facilitation is significantly enhanced, suggesting a reduction in the initial release probability of synaptic vesicles. Furthermore, Syb1-deficient NMJs display a pronounced asynchrony in neurotransmitter release. These impairments are not due to an alteration of the size of the readily releasable pool of vesicles, but are attributable to reduced sensitivity and cooperativity to calcium (Ca2+) due to the absence of Syb1. Our findings demonstrate that Syb1 plays an essential, non-redundant role in Ca2+-triggered vesicle exocytosis at the mouse NMJ.