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Reconstitution of Giant Mammalian Synapses in Culture for Molecular Functional and Imaging Studies
- Source :
- The Journal of Neuroscience. 36:3600-3610
- Publication Year :
- 2016
- Publisher :
- Society for Neuroscience, 2016.
-
Abstract
- Giant presynaptic terminal brain slice preparations have allowed intracellular recording of electrical signals and molecular loading, elucidating cellular and molecular mechanisms underlying neurotransmission and modulation. However, molecular genetic manipulation or optical imaging in these preparations is hampered by factors, such as tissue longevity and background fluorescence. To overcome these difficulties, we developed a giant presynaptic terminal culture preparation, which allows genetic manipulation and enables optical measurements of synaptic vesicle dynamics, simultaneously with presynaptic electrical signal recordings. This giant synapse reconstructed from dissociated mouse brainstem neurons resembles the development of native calyceal giant synapses in several respects. Thus, this novel preparation constitutes a powerful tool for studying molecular mechanisms of neurotransmission, neuromodulation, and neuronal development.SIGNIFICANCE STATEMENTWe have developed a novel culture preparation of giant mammalian synapses. These presynaptic terminals make it possible to perform optical imaging simultaneously with presynaptic electrophysiological recording. We demonstrate that this enables one to dissect endocytic and acidification times of synaptic vesicles. In addition, developmental elimination and functional maturation in this cultured preparation provide a useful model for studying presynaptic development. Because this giant synapse preparation allows molecular genetic manipulations, it constitutes a powerful new tool for studying molecular mechanisms of neurotransmission, neuromodulation, and neuronal development.
- Subjects :
- 0301 basic medicine
Endocytic cycle
Cell Culture Techniques
Action Potentials
Neurotransmission
Biology
Synaptic vesicle
Synapse
Mice
03 medical and health sciences
0302 clinical medicine
Slice preparation
Animals
Cells, Cultured
Microscopy
General Neuroscience
Brain
Articles
Neuromodulation (medicine)
Molecular Imaging
Electrophysiology
030104 developmental biology
Animals, Newborn
Synapses
Molecular imaging
Neuroscience
030217 neurology & neurosurgery
Subjects
Details
- ISSN :
- 15292401 and 02706474
- Volume :
- 36
- Database :
- OpenAIRE
- Journal :
- The Journal of Neuroscience
- Accession number :
- edsair.doi.dedup.....e3757b99a1e95b06c1e71b48327fb8c3