The mechanisms that determine how information is allocated to specific regions and cells in the brain are fundamentally important for memory capacity, storage and retrieval, but are poorly understood. Here, we manipulated CREB in a subset of lateral amygdala (LA) neurons with a modified Herpes Simplex Virus (HSV), and reversibly inactivated transfected neurons with the Drosophila allatostatin G-protein-coupled receptor (AlstR)/ligand system. We found that inactivation of the HSV-CREB subpopulation of neurons with allatostatin (AL) during training disrupted memory for tone conditioning, while inactivation of a similar proportion of HSV-LacZ control neurons did not. Whole-cell recordings of fluorescently tagged HSV-CREB neurons revealed that neurons with higher CREB levels are more excitable than neighboring neurons, and show larger synaptic efficacy changes following conditioning. Our findings demonstrate that CREB modulates the allocation of fear memory to specific cells in lateral amygdala, and suggest that neuronal excitability plays a key role in this process.

There is now compelling evidence that the allocation of memory to specific neurons (neuronal allocation) and synapses (synaptic allocation) in a neurocircuit is not random and that instead specific mechanisms, such as increases in neuronal excitability and synaptic tagging and capture, determine the exact sites where memories are stored. We propose an integrated view of these processes, such that neuronal allocation, synaptic tagging and capture, spine clustering and metaplasticity reflect related aspects of memory allocation mechanisms. Importantly, the properties of these mechanisms suggest a set of rules that profoundly affect how memories are stored and recalled.

Although memory allocation is a subject of active research in computer science, little is known about how the brain allocates information within neural circuits. There is an extensive literature on how specific types of memory engage different parts of the brain, and how neurons in these regions process and store information. Until recently, however, the mechanisms that determine how specific cells and synapses within a neural circuit (and not their neighbors) are recruited during learning have received little attention. Recent findings suggest that memory allocation is not random, but rather specific mechanisms regulate where information is stored within a neural circuit. Novel methods that allow tagging, imaging, activation and inactivation of neurons in behaving animals, promise to revolutionize studies of brain circuits, including memory allocation. Results from these studies are likely to have a considerable impact on both computer science as well as on the understanding of memory and its disorders.

The retrosplenial cortex (RSC) is part of a network of interconnected cortical, hippocampal, and thalamic structures harboring spatially modulated neurons. The RSC contains head direction cells and connects to the parahippocampal region and anterior thalamus. Manipulations of the RSC can affect spatial and contextual tasks. A considerable amount of evidence implicates the role of the RSC in spatial navigation, but it is unclear whether this structure actually encodes or stores spatial information. We used a transgenic mouse in which the expression of green fluorescent protein was under the control of the immediate early gene c-fos promoter as well as timelapse two-photon in vivo imaging to monitor neuronal activation triggered by spatial learning in the Morris water maze. We uncovered a repetitive pattern of cell activation in the RSC consistent with the hypothesis that during spatial learning an experience-dependent memory trace is formed in this structure. In support of this hypothesis, we also report three other observations. First, temporary RSC inactivation disrupts performance in a spatial learning task. Second, we show that overexpressing the transcription factor CREB in the RSC with a viral vector, a manipulation known to enhance memory consolidation in other circuits, results in spatial memory enhancements. Third, silencing the viral CREB-expressing neurons with the allatostatin system occludes the spatial memory enhancement. Taken together, these results indicate that the retrosplenial cortex engages in the formation and storage of memory traces for spatial information. S patial navigation, learning, and memory depend on several interconnected brain regions. The hippocampus is known to have a central role in spatial learning and memory. Place cells in this structure form a spatial map of an animal's environment (1). The main input to the hippocampus comes from the entorhinal cortex, an area that integrates cortical inputs before forwarding them into the hippocampal formation. The dorsal medial entorhinal cortex (MEC) contains spatially modulated cells (grid cells) that fire at the nodes of a hexagonal lattice as the animal traverses its environment (2). Beyond hippocampal place cells and MEC grid cells, there is also extensive evidence for another type of spatially modulated neurons that increase firing when the animal's head points in a specific direction [head direction (HD) cells] (3). These cells reside in several subcortical and cortical regions, including the retrosplenial cortex (RSC). The RSC is one of the most important cortical afferents to MEC (4, 5). It is the most caudal part of the cingulate cortex, positioned between anterior cingulate, parietal/visual, and parahippocampal cortices. It is further subdivided into areas A29a-c (granular cortex) and area A30 (dysgranular cortex) (6). The RSC has been implicated in a number of cognitive tasks, including navigation based on external spatial (allothetic) information (7). Human functional MRI (fMRI) studies showed that the RSC is activated duri...

During long-term memory formation, cellular and molecular processes reshape how individual neurons respond to specific patterns of synaptic input. It remains poorly understood how such changes impact information processing across networks of mammalian neurons. To observe how networks encode, store, and retrieve information, neuroscientists must track the dynamics of large ensembles of individual cells in behaving animals, over timescales commensurate with long-term memory. Fluorescence Ca2+-imaging techniques can monitor hundreds of neurons in behaving mice, opening exciting avenues for studies of learning and memory at the network level. Genetically encoded Ca2+ indicators allow neurons to be targeted by genetic type or connectivity. Chronic animal preparations permit repeated imaging of neural Ca2+ dynamics over multiple weeks. Together, these capabilities should enable unprecedented analyses of how ensemble neural codes evolve throughout memory processing and provide new insights into how memories are organized in the brain.

Recent findings suggest that memory allocation to specific neurons (i.e., neuronal allocation) in the amygdala is not random, but rather the transcription factor cAMP-response element binding protein (CREB) modulates this process, perhaps by regulating the transcription of channels that control neuronal excitability. Here, optogenetic studies in the mouse lateral amygdala (LA) were used to demonstrate that CREB and neuronal excitability regulate which neurons encode an emotional memory. To test the role of CREB in memory allocation, we overexpressed CREB in the lateral amygdala to recruit the encoding of an auditory-fear conditioning (AFC) memory to a subset of neurons. Then, post-training activation of these neurons with Channelrhodopsin-2 was sufficient to trigger recall of the memory for AFC, suggesting that CREB regulates memory allocation. To test the role of neuronal excitability in memory allocation, we used a step function opsin (SFO) to transiently increase neuronal excitability in a subset of LA neurons during AFC. Post-training activation of these neurons with Volvox Channelrhodopsin-1 was able to trigger recall of that memory. Importantly, our studies show that activation of the SFO did not affect AFC by either increasing anxiety or by strengthening the unconditioned stimulus. Our findings strongly support the hypothesis that CREB regulates memory allocation by modulating neuronal excitability.