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Reverse optical trawling for synaptic connections in situ.

Takuya Sasaki1, Genki Minamisawa, Naoya Takahashi

  • 1Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, Japan. ff077204@mail.ecc.u-tokyo.ac.jp

Journal of Neurophysiology
|April 24, 2009
PubMed
Summary

Researchers developed a novel method using functional multineuron calcium imaging to map neural network connectivity. This technique efficiently identifies presynaptic neurons by analyzing calcium transients and synaptic inputs, enhancing circuit mapping accuracy.

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Area of Science:

  • Neuroscience
  • Systems Neuroscience
  • Computational Neuroscience

Background:

  • Understanding neural circuit connectivity is crucial for deciphering brain function.
  • Existing methods for mapping synaptic connections are often limited in scale and resolution.
  • Characterizing the precise wiring of neuronal networks remains a significant challenge in neuroscience.

Purpose of the Study:

  • To introduce a novel, high-throughput method for mapping synaptic connectivity in neuronal networks.
  • To enhance the detection power and accuracy of identifying presynaptic neurons.
  • To provide a tool for studying the wiring topography of complex neuronal circuits.

Main Methods:

  • Whole-cell recording of postsynaptic neurons combined with functional multineuron calcium imaging of presynaptic neuron candidates.

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  • Statistical screening of neurons exhibiting calcium transients preceding postsynaptic inputs.
  • Optimization of extracellular ion concentrations ([K+]e, [Ca2+]e, [Mg2+]e) to improve synaptic transmission reliability.
  • Iontophoretic application of glutamate for targeted activation of presynaptic neurons ('trawling') to survey the network.

    • Main Results:

    • The developed method can identify presynaptic neurons synaptically connected to postsynaptic neurons.
    • Optimized ionic conditions and targeted activation significantly enhanced detection power.
    • The technique achieved a theoretical detection rate of 96% of activated presynaptic neurons with a 1% false-positive rate.
    • Demonstrated capability to map connectivity among dozens of neurons in brain slice preparations.

    Conclusions:

    • The introduced on-line probing technique is a powerful and accurate tool for mapping neural network connectivity.
    • This method significantly advances the ability to study the wiring topography of neuronal circuits.
    • The technique offers a promising approach for large-scale connectomics research in neuroscience.