New Approaches in Cognitive Neurobiology: Methods of Molecular Marking and Ex Vivo Imaging of Cognitively Active Neurons
- 17 Downloads
This review addresses the potentials of current methods of molecular ex vivo imaging of neurons involved in episodes of cognitive activity in experimental animals. We describe the principles on which the molecular identification of neurons activated in cognitive tasks are based, special attention being paid to the molecular marking of neuron activity in a single brain during two different cognitive episodes. Methods for double molecular labeling using in situ fluorescence hybridization (catFISH) are described in detail, along with approaches using transgenic animal strains to label neurons involved in cognitive activity via expression of fluorescent proteins within them (the tTA-tetO and TRAP Cre-loxP systems). The main advantages and disadvantages of these approaches are considered. Typical experimental schemes are presented which require specific aspects of working with these methods. A brief review of methods in which they are used to study the neural basis of cognitive activity in different behavioral tasks is presented. The potentials for the development of these approaches for studies of the cellular bases of higher brain functions are discussed.
Keywordscognitive activity learning memory brain neurons microscopy c-fos in situ hybridization catFISH transgene fluorescent protein tTA-tetO Cre-loxP
Unable to display preview. Download preview PDF.
- Broihier, H., “Whole-mount fluorescence in situ hybridization and antibody staining of Drosophila embryos,” Cold Spring Harb. Protoc., 7, No. 8, 900–905 (2012).Google Scholar
- Fu, T.-M., Guosong, H., Zhou, T., et al., “Stable long-term chronic brain mapping at the single-neuron level,” Nat. Methods, 8, 610–611 (2016).Google Scholar
- Guzowski, J. and Worley, P., “Cellular compartment analysis of temporal activity by fluorescence in situ hybridization (catFISH),” Curr. Protoc. Neuroscience, 1, 1–8 (2001).Google Scholar
- Hall, J., Thomas, K. L., and Everitt, B. J., “Cellular imaging of zif268 expression in the hippocampus and amygdala during contextual and cued fear memory retrieval: Selective activation of hippocampal CA1 neurons during the recall of contextual memories,” J. Neurosci., 21, 2186–2193 (2001).CrossRefGoogle Scholar
- Kaczmarek, L., “Gene expression in learning processes,” Acta Neurobiol. Exp., 60, 419–424 (2000).Google Scholar
- Roshchina, M. A., Ivashkina, O. I., and Anokhin, K. V., “New approaches in cognitive neurobiology: methods for the in vivo two-photon visualization of cognitively active neurons,” Zh. Vyssh. Nerv. Deyat., 67, No. 2, 1–9 (2017).Google Scholar
- Thomas, K., Hall, J., and Everitt, B., “Cellular imaging with zif268 expression in the rat nucleus accumbens and frontal cortex further dissociates the neural pathways activated following the retrieval of contextual and cued fear memory,” Eur. J. Neurosci., 16, 1789–1796 (2002).CrossRefGoogle Scholar
- Vazdarjanova, A., McNaughton, B., Barnes, C., et al., “Experience-dependent coincident expression of the effector immediate-early genes Arc and Homer 1a in hippocampal and neocortical neuronal networks,” J. Neurosci., 22, 10,067–10,071 (2002).Google Scholar