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Two-Photon Neurotransmitter Uncaging for the Study of Dendritic Integration

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Multiphoton Microscopy

Part of the book series: Neuromethods ((NM,volume 148))

Abstract

Neurons transform the information arising from up to thousands of synaptic inputs into specific spiking patterns. Nonlinear dendritic integration is a crucial step in this process and is thought to increase the computational ability of neurons. However, studying how complex spatiotemporal patterns of synaptic inputs drive neuronal spiking is technically challenging. Two-photon neurotransmitter uncaging allows researchers to activate sequences of single synapses with high spatiotemporal precision and thus systematically examine how single and multiple synaptic activation patterns may recruit dendritic nonlinearities. Here, we describe the theoretical and practical considerations of using two-photon uncaging to mimic synaptic activation and monitor the electrical and biochemical signaling in dendrites when evoked by various synaptic patterns.

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Abbreviations

[Glut]:

Glutamate concentration

2PLSM:

Two-photon laser scanning microscopy

ACSF:

Artificial cerebrospinal fluid

AMPAR:

α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor

AOD:

Acousto-optic deflector

AOM:

Acousto-optic modulator

CDNI-glutamate:

4-Carboxymethoxy-5,7-Dinitroindolinyl-Glutamate

dI/O:

Dendritic input/output relationship

DiO/DPA:

Two-component optical voltage sensor made of the neuronal tracer dye DiOC16(3) and dipicrylamine

DNI-glutamate TFA:

4-Methoxy-5,7-dinitroindolinyl-L-glutamate trifluoroacetate

GM:

The molecular two-photon cross-section is usually quoted in the units of Göppert-Mayer (GM), where 1 GM is 10−50 cm4 s photon−1

I/O:

Input/output relationship

MNI-Glutamate:

4-Methoxy-7-nitroindolinyl-caged-L-glutamate

NMDAR:

N-Methyl-D-aspartate receptor

PSF:

Point spread function

PV:

Parvalbumin

SLM:

Spatial light modulator

u[Glut]:

Uncaging-evoked glutamate transient

uEPSC:

Uncaging-evoked EPSC

uEPSP:

Uncaging-evoked EPSP

VGCC:

Voltage-gated calcium channel

θ B :

Bragg angle, the angle between the incident laser beam into an AOM and the diffracted beam

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Authors and Affiliations

  1. Laboratory of Dynamic Neuronal Imaging, Department of Neuroscience, Institut Pasteur, Paris, France

    Alexandra Tran-Van-Minh, Nelson Rebola, Andreas Hoehne & David A. DiGregorio

  2. CNRS UMR 3571, Paris, France

    Alexandra Tran-Van-Minh, Nelson Rebola, Andreas Hoehne & David A. DiGregorio

Authors
  1. Alexandra Tran-Van-Minh
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  2. Nelson Rebola
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  3. Andreas Hoehne
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  4. David A. DiGregorio
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Correspondence to David A. DiGregorio .

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Editors and Affiliations

  1. Department of Biomedicine, University of Bergen, Bergen, Norway

    Espen Hartveit

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Tran-Van-Minh, A., Rebola, N., Hoehne, A., DiGregorio, D.A. (2019). Two-Photon Neurotransmitter Uncaging for the Study of Dendritic Integration. In: Hartveit, E. (eds) Multiphoton Microscopy. Neuromethods, vol 148. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9702-2_3

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  • DOI: https://doi.org/10.1007/978-1-4939-9702-2_3

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