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Updated: Aug 21, 2025

Two-photon Calcium Imaging in Neuronal Dendrites in Brain Slices

Two-photon Calcium Imaging in Neuronal Dendrites in Brain Slices

Published on: March 15, 2018

Fast Synaptically Activated Calcium and Sodium Kinetics in Hippocampal Pyramidal Neuron Dendritic Spines.

Kenichi Miyazaki¹, William N Ross²

¹Department of Physiology, New York Medical College, Valhalla, NY 10595.

|November 15, 2022

Summary

New research reveals that sodium influx in hippocampal neuron spines primarily uses AMPA receptors, with varying decay times indicating diverse spine neck resistances. Calcium influx is rapid, mainly via NMDA receptors, differing between synaptic release and glutamate uncaging.

Keywords:

AMPA NMDA calcium sodium spine

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Last Updated: Aug 21, 2025

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Published on: May 25, 2011

Area of Science:

Neuroscience
Cellular Neuroscience
Synaptic Physiology

Background:

Accurate assessment of postsynaptic currents is crucial for understanding synaptic integration and signaling in dendritic spines.
Previous studies using two-photon imaging and uncaging have provided insights but left gaps in understanding the time courses of synaptically evoked intracellular calcium ([Ca2+]i) and sodium ([Na+]i) changes.

Purpose of the Study:

To extend measurements of spine parameters, specifically the time courses of synaptically evoked [Ca2+]i and [Na+]i changes.
To investigate the primary sources of Na+ and Ca2+ influx in CA1 hippocampal pyramidal neuron spines.

Main Methods:

Utilized low-affinity, linear Na+ and Ca2+ indicators.
Employed laser fluorescence stimulation and sensitive camera-based detection.
Combined with electrical stimulation and two-photon glutamate uncaging.

Main Results:

Synaptically activated Na+ currents in spines are predominantly mediated by AMPA receptors, with minimal entry via voltage-gated sodium channels (VGSCs) or NMDA receptor channels.
Observed a spectrum of sodium transient decay times, suggesting variations in spine neck resistance.
Synaptically activated [Ca2+]i changes are rapid and primarily result from NMDA receptor Ca2+ influx when the Mg2+ block is relieved.
Glutamate uncaging evoked slower [Ca2+]i changes compared to synaptic release, indicating a greater contribution of resting NMDA receptor Ca2+ influx after uncaging.

Conclusions:

Synaptic Na+ influx in CA1 pyramidal neuron spines is mainly through AMPA receptors.
Dendritic spines exhibit diverse neck resistances influencing sodium transient dynamics.
NMDA receptor activation is critical for synaptically evoked Ca2+ transients, with timing dependent on the method of stimulation (synaptic release vs. uncaging).