Learning and Memory

Visualization of neuronal network activity and plasticity 

During learning and memory formation, gene expression is dynamically regulated in response to experience-dependent neuronal activity. In particular, the expression of immediate-early genes (IEGs) including c-fos, and Arc is rapidly and transiently altered in specific neurons or neuronal ensembles in brain areas involved in the learning and memory processing of a certain task/activity. Therefore, the visualization of IEG mRNA expression patterns have widely been used as a molecular readout for neuronal populations that are engaged in generating and encoding long-term memories. 

To further characterize activated neuronal cell populations, RNAscope® multiplexing capability enables IEG mRNA detection in combination with other targets or cell type markers. 

Here, we detect neuronal activity in the mouse brain hippocampus by using the RNAscope® assay to visualize Cfos which is a proto-oncogene and transcription factor that is induced within 15 minutes of stimulation (Figure 1). Simultaneously, we used a second probe to detect Chrm3-positive neurons.


FIGURE 1. Detection of the transcription factor immediate early gene and activity marker Cfos (Green) in combination with the G protein-coupled receptor Cholinergic Receptor Muscarinic 3 (Chrm3, Red) in normal mouse brain hippocampus using the RNAscope® Multiplex Fluorescent assay on Fresh Frozen tissue samples. Cells are counterstained with DAPI. CA = Cornu Ammonis.

To detect neuronal plasticity, the expression of the effector immediate early gene Activity-Regulated Cytoskeleton-associated protein Arc (also known as Arg3.1) is visualized in Figure 2 in combination with a probe targeted to Drd1.


FIGURE 2. (Right panel) Detection of the effector immediate early gene and plasticity marker Arc (Green) in combination with the G protein-coupled Dopamine Receptor D1 (Drd1, Red) in normal mouse brain striatum using the RNAscope® Multiplex Fluorescent assay on Fresh Frozen tissue samples. (Top left panel) Single-channel image for Drd1 (Red). (Bottom left panel) Single-channel image for Arc (Green).

References:​

  • Guzowski JF. Insights into immediate-early gene function in hippocampal memory consolidation using antisense oligonucleotide and fluorescent imaging approaches. Hippocampus 2002; 12:86-104

Application Note

Learn more in our RNAscope® Neuroscience Application Note - Detection of RNA in the central and peripheral nervous system using the RNAscope® in situ hybridization assay

Detection of RNA in the central and peripheral nervous system using the RNAscope® in situ hybridization assay

Fill out the form below to download the application note:

Complete form to download PDF

Neuroscience Gene List

(click on gene name to access corresponding RNAscope® probes)

 

 

 

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