New Tools in Cognitive Neurobiology: Biotin-Digoxigenin Detection of Overlapping Active Neuronal Populations by Two-Color c-fos Compartment Analysis of Temporal Activity by Fluorescent in situ Hybridization (catFISH)

Saidov, Khalid; Тиунова, А. А.; Subach, Fedor V.; Anokhin, Konstantin V.

doi:10.21926/obm.genet.1901065

Cited by 1 publication

(4 citation statements)

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“…In addition to Fos activity imaging, we used Fos-associated transgenic mouse approach as a proof-of-principle for long-term investigation of calcium activity in neurons specifically tagged during a cognitive episode. Currently, different IEG-based methods such as compartment analysis of temporal activity by fluorescent in-situ hybridization (catFISH) or TetTag and TRAP transgenic systems are used to label histochemically neurons that were activated during two behavioral episodes [ 25 , 32 , 33 , 35 , 36 ]. However, the catFISH technique has a limited tagging window that allows comparing populations of neurons that were activated in two episodes with only about 30 min time window [ 36 ].…”

Section: Discussionmentioning

confidence: 99%

“…Currently, different IEG-based methods such as compartment analysis of temporal activity by fluorescent in-situ hybridization (catFISH) or TetTag and TRAP transgenic systems are used to label histochemically neurons that were activated during two behavioral episodes [ 25 , 32 , 33 , 35 , 36 ]. However, the catFISH technique has a limited tagging window that allows comparing populations of neurons that were activated in two episodes with only about 30 min time window [ 36 ]. Also, catFISH is not suitable for the investigation of the dynamic activity of labeled cells.…”

Section: Discussionmentioning

confidence: 99%

“…One possibility relates to the laminar heterogeneity of neocortex involvement in memory storage [ 24 , 37 ]. In our experiments we used memory retrieval test 24 h after training, a period which qualifies as a recent memory [ 24 , 25 , 36 ]. Notably, consolidation from the recent to remote long-term memory was shown to be accompanied by a laminar reorganization of neuronal activity in the mouse parietal cortex [ 24 ].…”

Section: Discussionmentioning

confidence: 99%

“…Due to limitations of two-photon imaging, here, we were restricted to sampling only superficial layers of the PtA. Neurons from these layers might be less involved in the recent memory retrieval compared to deeper layer cortical neurons as was previously shown for other cortical areas [ 24 , 36 , 37 ]. This hypothesis can be tested experimentally by using GRIN(Gradient-Index)-lens two-photon imaging, three-photon imaging, or miniscopes to examine the laminar distribution of Fos-Cre-GCaMP neurons responsive to CS presentation.…”

Section: Discussionmentioning

confidence: 99%

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Imaging of C-fos Activity in Neurons of the Mouse Parietal Association Cortex during Acquisition and Retrieval of Associative Fear Memory

Ivashkina

Gruzdeva

Roshchina

et al. 2021

IJMS

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The parietal cortex of rodents participates in sensory and spatial processing, movement planning, and decision-making, but much less is known about its role in associative learning and memory formation. The present study aims to examine the involvement of the parietal association cortex (PtA) in associative fear memory acquisition and retrieval in mice. Using ex vivo c-Fos immunohistochemical mapping and in vivo Fos-EGFP two-photon imaging, we show that PtA neurons were specifically activated both during acquisition and retrieval of cued fear memory. Fos immunohistochemistry revealed specific activation of the PtA neurons during retrieval of the 1-day-old fear memory. In vivo two-photon Fos-EGFP imaging confirmed this result and in addition detected specific c-Fos responses of the PtA neurons during acquisition of cued fear memory. To allow a more detailed study of the long-term activity of such PtA engram neurons, we generated a Fos-Cre-GCaMP transgenic mouse line that employs the Targeted Recombination in Active Populations (TRAP) technique to detect calcium events specifically in cells that were Fos-active during conditioning. We show that gradual accumulation of GCaMP3 in the PtA neurons of Fos-Cre-GCaMP mice peaks at the 4th day after fear learning. We also describe calcium transients in the cell bodies and dendrites of the TRAPed neurons. This provides a proof-of-principle for TRAP-based calcium imaging of PtA functions during memory processes as well as in experimental models of fear- and anxiety-related psychiatric disorders and their specific therapies.

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