Stimulation of Exocytosis in Cultured Cerebellar Granule Cells by Electrical Field Stimulation

Cousin, Michael A.; Hurst, Helen C.; Held, Brigitte; Nicholls, David G.

doi:10.1007/978-3-642-60704-2_7

Cited by 2 publications

(4 citation statements)

References 84 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…rons (Wu and Betz, 1996;Cousin and Nicholls, 1997;Cousin et al, 1997a). Similarly, the magnitude of fluorescence loss from nerve terminals loaded with styryl dyes of different hydrophobicity (which have a different t 1/2 on the vesicle membrane) was identical at low stimulation frequencies in cultured hippocampal neurons (Ryan et al, 1996a).…”

Section: Full or Transient Vesicle Fusion?mentioning

confidence: 87%

“…However, changes in stimulus intensity do not affect the mechanism of synaptic vesicle fusion. For example, no difference was observed between the extent of styryl dye release and neurotransmitter release with varying stimulation frequency in several types of neurons (Wu and Betz, 1996 ; Cousin and Nicholls, 1997 ; Cousin et al, 1997 a ). Similarly, the magnitude of fluorescence loss from nerve terminals loaded with styryl dyes of different hydrophobicity (which have a different t 1/2 on the vesicle membrane) was identical at low stimulation frequencies in cultured hippocampal neurons (Ryan et al, 1996 a ).…”

Section: Synaptic Vesicle Fusionmentioning

confidence: 96%

“…For example, no difference in the kinetics of dye loss in hippocampal neurons was found from the readily releasable vesicle pool whether it was fully or partially loaded with FM1‐43 (Ryan and Smith, 1995). In primary cultures of cerebellar granule cells, the stimulated loss of styryl dyes was halved with every train of action potential stimuli (Cousin and Nicholls, 1997 ; Cousin et al, 1997 a ). This was not due to depletion of the readily releasable vesicle pool, because the release of soluble vesicle contents remained constant, but indicates that the dye‐labelled synaptic vesicles were being diluted by new unlabelled vesicles after every stimulus.…”

Section: Vesicle Traffic : Routes and Regulationmentioning

confidence: 99%

See 2 more Smart Citations

Mechanisms of Synaptic Vesicle Recycling Illuminated by Fluorescent Dyes

Cousin

Robinson

1999

Journal of Neurochemistry

Self Cite

View full text Add to dashboard Cite

Abstract:The recycling of synaptic vesicles in nerve terminals involves multiple steps, underlies all aspects of synaptic transmission, and is a key to understanding the basis of synaptic plasticity. The development of styryl dyes as fluorescent molecules that label recycling synaptic vesicles has revolutionized the way in which synaptic vesicle recycling can be investigated, by allowing an examination of processes in neurons that have long been inaccessible. In this review, we evaluate the major aspects of synaptic vesicle recycling that have been revealed and advanced by studies with styryl dyes, focussing upon synaptic vesicle fusion, retrieval, and trafficking. The greatest impact of styryl dyes has been to allow the routine visualization of endocytosis in central nerve terminals for the first time. This has revealed the kinetics of endocytosis, its underlying sequential steps, and its regulation by Ca 2ϩ . In studies of exocytosis, styryl dyes have helped distinguish between different modes of vesicle fusion, provided direct support for the quantal nature of exocytosis and endocytosis, and revealed how the probability of exocytosis varies enormously from one nerve terminal to another. Synaptic vesicle labelling with styryl dyes has helped our understanding of vesicle trafficking by allowing better understanding of different synaptic vesicle pools within the nerve terminal, vesicle intermixing, and vesicle clustering at release sites. Finally, the dyes are now being used in innovative ways to reveal further insights into synaptic plasticity.

show abstract

Section: Full or Transient Vesicle Fusion?mentioning

confidence: 87%

Section: Synaptic Vesicle Fusionmentioning

confidence: 96%

Section: Vesicle Traffic : Routes and Regulationmentioning

confidence: 99%

See 1 more Smart Citation

Mechanisms of Synaptic Vesicle Recycling Illuminated by Fluorescent Dyes

Cousin

Robinson

1999

Journal of Neurochemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…[ 3 H]‐ d ‐aspartate release from cultured cerebellar granule cells was determined as described previously (Cousin et al . 1997;Varming et al .…”

Section: Methodsmentioning

confidence: 99%

Modulation of neurotransmitter release by dihydropyridine‐sensitive calcium channels involves tyrosine phosphorylation

Evans

Pocock

1999

Eur J of Neuroscience

View full text Add to dashboard Cite

Cultured rat cerebellar granule cells depolarized by high KCl, display a large component of Ca2+ influx through L-type voltage-dependent Ca2+ channels as defined by a sensitivity to 1 microM nifedipine. This Ca2+ influx is not coupled to neurotransmitter exocytosis but has implications for neuronal development. KCl stimulation in the absence of external Ca2+ followed by the readdition of Ca2+ allows the coupling of a class of L-type Ca2+ channels to neurotransmitter exocytosis as assessed by loading of glutamatergic pools with [3H]-D-aspartate. KCl stimulation in the absence of external Ca2+ ('predepolarization') enhances tyrosine phosphorylation of several cellular proteins, and inhibitors of tyrosine kinases block both phosphorylation and the neurotransmitter release coupled to the L-type Ca2+ channel. More specifically, an inhibitor of src family tyrosine kinases, PP1, blocks the effects of predepolarization suggesting a role for a src family kinase in the process. Furthermore, L-type Ca2+ channel recruitment and modulation of release could be activated with the tyrosine phosphatase inhibitor sodium orthovanadate. The phosphoproteins enhanced by predepolarization, which include the cytoskeletal proteins focal adhesion kinase (FAK) and vinculin, are also highly phosphorylated early on in culture when neurite outgrowth occurs. As the neurons develop a network of neurites, both tyrosine phosphorylation and L-type Ca2+ channel activity decrease. These results show a novel mechanism for the recruitment of L-type Ca2+ channels and their coupling to neurotransmitter release which involves tyrosine phosphorylation. This phenomenon has a role in cerebellar granule cell development.

show abstract

Stimulation of Exocytosis in Cultured Cerebellar Granule Cells by Electrical Field Stimulation

Cited by 2 publications

References 84 publications

Mechanisms of Synaptic Vesicle Recycling Illuminated by Fluorescent Dyes

Mechanisms of Synaptic Vesicle Recycling Illuminated by Fluorescent Dyes

Modulation of neurotransmitter release by dihydropyridine‐sensitive calcium channels involves tyrosine phosphorylation

Contact Info

Product

Resources

About