Abstract
To facilitate a functional analysis of neuronal connectivity in a mammalian nervous system that is tightly packed with billions of cells, we developed a new technique that uses inducible genetic manipulations in fluorescently labeled single neurons in mice. Our technique, single-neuron labeling with inducible Cre-mediated knockout (SLICK), is achieved by coexpressing a drug-inducible form of Cre recombinase and a fluorescent protein in a small subsets of neurons, thus combining the powerful Cre recombinase system for conditional genetic manipulation with fluorescent labeling of single neurons for imaging. Here, we demonstrate efficient inducible genetic manipulation in several types of neurons using SLICK. Furthermore, we applied SLICK to eliminate synaptic transmission in a small subset of neuromuscular junctions. Our results provide evidence for the long-term stability of inactive neuromuscular synapses in adult animals and demonstrate a Cre-loxP compatible system for dissecting gene functions in single identifiable neurons.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Central Nervous System / cytology
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Estrogen Antagonists / pharmacology
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Ganglia, Spinal / cytology
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Gene Expression / drug effects
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Gene Expression / genetics
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Genes, Reporter / physiology
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Green Fluorescent Proteins / biosynthesis
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Green Fluorescent Proteins / genetics
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Integrases / genetics
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Integrases / metabolism*
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Luminescent Proteins / biosynthesis
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Luminescent Proteins / genetics
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Mice
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Mice, Inbred C57BL
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Mice, Knockout / anatomy & histology
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Mice, Knockout / metabolism*
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Mice, Transgenic
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Nerve Tissue Proteins / deficiency
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Nerve Tissue Proteins / genetics
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Neurons / classification
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Neurons / cytology
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Neurons / metabolism*
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Potassium Channels / genetics
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Potassium Channels / metabolism
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Potassium Channels, Sodium-Activated
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Tamoxifen / pharmacology
Substances
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Estrogen Antagonists
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Kcnt2 protein, mouse
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Luminescent Proteins
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Nerve Tissue Proteins
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Potassium Channels
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Potassium Channels, Sodium-Activated
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Tamoxifen
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Green Fluorescent Proteins
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Cre recombinase
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Integrases