Simultaneous electrophysiological recording and calcium imaging of suprachiasmatic nucleus neurons.

Robert P. Irwin, Charles Allen

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in regulating the intracellular calcium concentration. Changing environmental conditions, such as the light-dark cycle, can modify neuronal and neural network activity and the expression of a family of circadian clock genes within the suprachiasmatic nucleus (SCN), the location of the master circadian clock in the mammalian brain. Excitatory synaptic transmission leads to an increase in the postsynaptic Ca(2+) concentration that is believed to activate the signaling pathways that shifts the rhythmic expression of circadian clock genes. Hypothalamic slices containing the SCN were patch clamped using microelectrodes filled with an internal solution containing the calcium indicator bis-fura-2. After a seal was formed between the microelectrode and the SCN neuronal membrane, the membrane was ruptured using gentle suction and the calcium probe diffused into the neuron filling both the soma and dendrites. Quantitative ratiometric measurements of the intracellular calcium concentration were recorded simultaneously with membrane potential or current. Using these methods it is possible to study the role of changes of the intracellular calcium concentration produced by synaptic activity and action potential firing of individual neurons. In this presentation we demonstrate the methods to simultaneously record electrophysiological activity along with intracellular calcium from individual SCN neurons maintained in brain slices.

Original languageEnglish (US)
Pages (from-to)50794
Number of pages1
JournalJournal of visualized experiments : JoVE
Issue number82
StatePublished - 2013
Externally publishedYes

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Suprachiasmatic Nucleus
Neurons
Calcium
Imaging techniques
Circadian Clocks
Membranes
Clocks
Microelectrodes
Membrane Potentials
Brain
Genes
Synaptic Potentials
Fura-2
Photoperiod
Suction
Carisoprodol
Dendrites
Synaptic Transmission
Action Potentials
Seals

ASJC Scopus subject areas

  • Medicine(all)

Cite this

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