Molecular and genetic determinants of the NMDA receptor for superior learning and memory functions.

The opening-duration of the NMDA receptors implements Hebb's synaptic coincidence-detection and is long thought to be the rate-limiting factor underlying superior memory. Here, we investigate the molecular and genetic determinants of the NMDA receptors by testing the "synaptic coincidence-detection time-duration" hypothesis vs. "GluN2B intracellular signaling domain" hypothesis. Accordingly, we generated a series of GluN2A, GluN2B, and GluN2D chimeric subunit transgenic mice in which C-terminal intracellular domains were systematically swapped and overexpressed in the forebrain excitatory neurons. The data presented in the present study supports the second hypothesis, the "GluN2B intracellular signaling domain" hypothesis. Surprisingly, we found that the voltage-gated channel opening-durations through either GluN2A or GluN2B are sufficient and their temporal differences are marginal. In contrast, the C-terminal intracellular domain of the GluN2B subunit is necessary and sufficient for superior performances in long-term novel object recognition and cued fear memories and superior flexibility in fear extinction. Intriguingly, memory enhancement correlates with enhanced long-term potentiation in the 10-100 Hz range while requiring intact long-term depression capacity at the 1-5 Hz range.

Increased NR2A:NR2B ratio compresses long-term depression range and constrains long-term memory.

The NR2A:NR2B subunit ratio of the NMDA receptors is widely known to increase in the brain from postnatal development to sexual maturity and to aging, yet its impact on memory function remains speculative. We have generated forebrain-specific NR2A overexpression transgenic mice and show that these mice had normal basic behaviors and short-term memory, but exhibited broad long-term memory deficits as revealed by several behavioral paradigms. Surprisingly, increased NR2A expression did not affect 1-Hz-induced long-term depression (LTD) or 100 Hz-induced long-term potentiation (LTP) in the CA1 region of the hippocampus, but selectively abolished LTD responses in the 3-5 Hz frequency range. Our results demonstrate that the increased NR2A:NR2B ratio is a critical genetic factor in constraining long-term memory in the adult brain. We postulate that LTD-like process underlies post-learning information sculpting, a novel and essential consolidation step in transforming new information into long-term memory.

Differential consolidation and pattern reverberations within episodic cell assemblies in the mouse hippocampus.

One hallmark feature of consolidation of episodic memory is that only a fraction of original information, which is usually in a more abstract form, is selected for long-term memory storage. How does the brain perform these differential memory consolidations? To investigate the neural network mechanism that governs this selective consolidation process, we use a set of distinct fearful events to study if and how hippocampal CA1 cells engage in selective memory encoding and consolidation. We show that these distinct episodes activate a unique assembly of CA1 episodic cells, or neural cliques, whose response-selectivity ranges from general-to-specific features. A series of parametric analyses further reveal that post-learning CA1 episodic pattern replays or reverberations are mostly mediated by cells exhibiting event intensity-invariant responses, not by the intensity-sensitive cells. More importantly, reactivation cross-correlations displayed by intensity-invariant cells encoding general episodic features during immediate post-learning period tend to be stronger than those displayed by invariant cells encoding specific features. These differential reactivations within the CA1 episodic cell populations can thus provide the hippocampus with a selection mechanism to consolidate preferentially more generalized knowledge for long-term memory storage.

CaMKII activation state underlies synaptic labile phase of LTP and short-term memory formation.

BACKGROUND: The labile state of short-term memory has been known for more than a century. It has been frequently reported that immediate postlearning intervention can readily disrupt newly formed memories. However, the molecular and cellular mechanisms underlying the labile state of new memory are not understood. RESULTS: Using a bump-and-hole-based chemical-genetic method, we have rapidly and selectively manipulated alpha CaMKII activity levels in the mouse forebrain during various stages of the short-term memory processes. We find that a rapid shift in the alpha CaMKII activation status within the immediate 10 min after learning severely disrupts short-term memory formation. The same manipulation beyond the 15 min after learning has no effect, suggesting a critical time window for CaMKII action. We further show that during this same 10 min time window only, shifting in CaMKII activation state is capable of altering newly established synaptic weights and/or patterns. CONCLUSION: The initial 10 min of memory formation and long-term potentiation are sensitive to inducible genetic upregulation of alphaCaMKII activity. Our results suggest that molecular dynamics of CaMKII play an important role in underlying synaptic labile state and representation of short-term memory during this critical time window.

Maintenance of superior learning and memory function in NR2B transgenic mice during ageing.

Brain ageing represents a general and evolutionarily conserved phenomenon and is marked by gradual declines in cognitive functions such as learning and memory. As a synaptic coincidence detector, the N-methyl-d-aspartate (NMDA) receptor is known to be essential for the induction of synaptic plasticity and memory formation. Here, we test the hypothesis that up-regulation of NR2B expression is beneficial for learning and memory in the aged animals. Our in vitro recordings show that the aged transgenic mice with the forebrain-specific overexpression of the NR2B subunit indeed exhibit more robust hippocampal long-term potentiation (LTP) induced by either high-frequency stimulation or theta-stimulation protocol. Furthermore, those aged NR2B transgenic mice consistently outperform their wild-type littermates in five different learning and memory tests, namely, novel object recognition, contextual and cued fear conditioning, spatial reference memory, and spatial working memory T-maze task. Thus, we conclude that increased expression of NR2B in the forebrain improves learning and memory function in the aged brain.

Forebrain degeneration and ventricle enlargement caused by double knockout of Alzheimer's presenilin-1 and presenilin-2.

Early-onset familial Alzheimer's disease is the most aggressive form of Alzheimer's, striking patients as early as their 30s; those patients typically carry mutations in presenilin-1 and presenilin-2. To investigate the coordinated functions of presenilin in the adult brain, we generated double knockout mice, in which both presenilins were deleted in the forebrain. We found that concurrent loss of presenilins in adulthood resulted in massive cortical shrinkage, atrophy of hippocampal molecular layers and corpus callosum, and enlargement of the lateral and third ventricles. We further revealed that deficiency of presenilins caused a series of biochemical alterations, including neuronal atrophy, astrogliosis, caspase-3-mediated apoptosis, and tau hyperphosphorylation. Thus, our study demonstrates that presenilins are essential for the ongoing maintenance of cortical structures and function.

GABAergic inhibition and the effect of sound direction on rate-intensity functions of inferior collicular neurons of the big brown Bat, Eptesicus fuscus.

GABAergic inhibition shapes many auditory response properties of neurons in the inferior colliculus of the big brown bat, Eptesicus fuscus. This study examined the role of GABAergic inhibition on direction-dependent rate-intensity functions of bat inferior collicular neurons. When plotted at three sound directions (60 degrees contralateral, 0 degrees and 60 degrees ipsilateral relative to recording site), most collicular neurons had nonmonotonic and saturated rate-intensity functions at 60 degrees contralateral and 0 degrees but had monotonic rate-intensity functions at 60 degrees ipsilateral. The dynamic range of rate-intensity functions of majority (>90%) of collicular neurons significantly decreased as the sound direction changed from 60 degrees contralateral to 60 degrees ipsilateral. Bicuculline application increased or decreased the dynamic range of IC neurons in different degrees with sound direction and abolished direction-dependent intensity sensitivity of these IC neurons. Possible mechanisms for these observations are discussed.

Inducible protein knockout reveals temporal requirement of CaMKII reactivation for memory consolidation in the brain.

By integrating convergent protein engineering and rational inhibitor design, we have developed an in vivo conditional protein knockout andor manipulation technology. This method is based on the creation of a specific interaction interface between a modified protein domain and sensitized inhibitors. By introducing this system into genetically modified mice, we can readily manipulate the activity of a targeted protein, such as alpha-Ca(2+)calmodulin-dependent protein kinase II (alphaCAMKII), on the time scale of minutes in specific brain subregions of freely behaving mice. With this inducible and region-specific protein knockout technique, we analyzed the temporal stages of memory consolidation process and revealed the first postlearning week as the critical time window during which a precise level of CaMKII reactivation is essential for the consolidation of long-term memories in the brain.