Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins

@article{Miesenbck1998VisualizingSA,
  title={Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins},
  author={G. Miesenb{\"o}ck and Dino A. De Angelis and J. Rothman},
  journal={Nature},
  year={1998},
  volume={394},
  pages={192-195}
}
In neural systems, information is often carried by ensembles of cells rather than by individual units. Optical indicators provide a powerful means to reveal such distributed activity, particularly when protein-based and encodable in DNA: encodable probes can be introduced into cells, tissues, or transgenic organisms by genetic manipulation, selectively expressed in anatomically or functionally defined groups of cells, and, ideally, recorded in situ, without a requirement for exogenous cofactors… Expand
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References

SHOWING 1-10 OF 29 REFERENCES
Patterns of synaptic activity in neural networks recorded by light emission from synaptolucins.
  • G. Miesenböck, J. Rothman
  • Biology, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
  • 1997
TLDR
Fusion proteins of Cypridina luciferase and synaptotagmin-I or VAMP-2/synaptobrevin were expressed in cultured hippocampal neurons with the help of viral vectors and formed light-emitting complexes with their cognate luciferin, which was added to the extracellular medium. Expand
Fluorescent indicators for Ca2+based on green fluorescent proteins and calmodulin
TLDR
New fluorescent indicators for Ca2+ that are genetically encoded without cofactors and are targetable to specific intracellular locations are constructed and dubbed ‘cameleons’. Expand
Fluorescent probes of cell signaling.
  • R. Tsien
  • Chemistry, Medicine
  • Annual review of neuroscience
  • 1989
TLDR
The earliest developed and most straight­ forward uses of fluorescent groups are simply as positional tags or markers, while a second type of application of fluorescent probes involves attachment of the fluorophore to a purified macromolecule to sense conformational change of the latter. Expand
Wavelength mutations and posttranslational autoxidation of green fluorescent protein.
  • R. Heim, D. Prasher, R. Tsien
  • Biology, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
  • 1994
TLDR
The availability of two visibly distinct colors should significantly extend the usefulness of GFP in molecular and cell biology by enabling in vivo visualization of differential gene expression and protein localization and measurement of protein association by fluorescence resonance energy transfer. Expand
Cellubrevin is a ubiquitous tetanus-toxin substrate homologous to a putative synaptic vesicle fusion protein
TLDR
The results suggest that constitutive and regulated vesicular pathways use homologous proteins for membrane trafficking, probably for membrane fusion at the plasma membrane, indicating a greater mechanistic and evolutionary similarity between these pathways than previously thought. Expand
Capacitance measurements reveal stepwise fusion events in degranulating mast cells
TLDR
The results show that degranulation occurs spontaneously and reproducibly if the GTP analogue, GTP-γ-S, and Mg-ATP are present in the pipette filling solution and that guanine nucleotide regulatory proteins are involved in the control of this process. Expand
Detection in Living Cells of Ca2+-dependent Changes in the Fluorescence Emission of an Indicator Composed of Two Green Fluorescent Protein Variants Linked by a Calmodulin-binding Sequence
TLDR
This observation suggests that the activity of a calmodulin target with a typical 1 nmaffinity for (Ca2+)4-calmodulin is responsive to changes in the intracellular Ca2+ concentration over the physiological range. Expand
Mutations that suppress the thermosensitivity of green fluorescent protein
TLDR
The thermotolerant mutants of GFP greatly improve the sensitivity of the protein as a visible reporter molecule in bacterial, yeast and mammalian cells and produces a thermostable folding mutant (GFP5) that can be efficiently excited using either long-wavelength ultraviolet or blue light. Expand
SPECTRAL PERTURBATIONS OF THE AEQUOREA GREEN‐FLUORESCENT PROTEIN
Abstract— In the jellyfish Aequorea, the green‐fluorescent protein (GFP) functions as the in vivo bio‐luminescence emitter via energy transfer from the photoprotein aequorin. Accumulated evidence hasExpand
Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer
TLDR
The results demonstrate that the production of more and better GFP variants is possible and worthwhile, and facilitates multicolor imaging of differential gene expression, protein localization or cell fate. Expand
...
1
2
3
...