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J Neurosci Methods. 2019 Jul 26;326:108368. doi: 10.1016/j.jneumeth.2019.108368. [Epub ahead of print]

Single-cell memory trace imaging with immediate-early genes.

Author information

1
Leibniz-Institute for Neurobiology, Functional Architecture of Memory Dept., 39118, Magdeburg, Germany; Otto von Guericke University, Medical Faculty, Functional Neuroplasticity Dept., 39120, Magdeburg, Germany; Otto von Guericke University, Center for Behavioral Brain Sciences, 39106, Magdeburg, Germany. Electronic address: magdalena.sauvage@lin-magdeburg.de.
2
Graduate school of Frontier Biosciences, Osaka University, Suita, Osaka, 565-0871, Japan.
3
Leibniz-Institute for Neurobiology, Functional Architecture of Memory Dept., 39118, Magdeburg, Germany.

Abstract

For the past decades, an increasing number of studies has taken advantage of molecular imaging methods involving the detection of immediate-early genes' (IEGs) expression for investigating neural substrates underlying plasticity processes and memory function. The detection of IEGs RNA by Fluorescent In-Situ Hybridization (FISH) yields single-cell as well as high temporal resolution and has recently enabled the mapping of medial temporal lobe subareas/subnetworks activity induced by single or multiple behavioural events in the same animal. After briefly reviewing the function and the ties of the typical IEGs (Fos, Zif268, Arc, Homer1a) used for mapping plasticity, we focus on discussing technical considerations vital for the successful detection of IEGs with FISH with emphasis on the design of RNA probes, the optimization of experimental conditions and the necessity for controls. Finally, we discuss recent developments in brain clearing methods that in combination with FISH detection of IEGs' expression allow for 3D imaging with single cell resolution as well as whole brain analyses. This, in parallel with the recent development of fMRI cognitive tasks in awake rats and the use of high resolution fMRI in humans, holds great promises for bridging further memory in humans and animals.

KEYWORDS:

Arc; Catfish; FISH; IEG; Immediate-early-gene; Memory

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