The structures of the medial temporal lobe (MTL) have been shown to be causally involved in episodic and recognition memory. However, recent work in a number of species has demonstrated that impairments in recognition memory seen following lesions of the perirhinal cortex can be accounted for by deficits in perceptual discrimination. These findings suggest that object representation, rather than explicit recognition memory signals, may be crucial to the mnemonic process. Given the large amount of visual information encountered by primates, there must be a reconsideration of the mechanisms by which the brain efficiently stores visually presented information. Previous neurophysiological recordings from MTL structures in primates have largely focused on tasks that implicitly define object familiarity (i.e., novel vs. familiar) or contain significant mnemonic demands (e.g., conditional associations between two stimuli), limiting their utility in understanding the mechanisms underlying visual object recognition and information storage. To clarify how different regions in the MTL may contribute to visual recognition we recorded from three rhesus macaques performing a passive viewing task. The task design systematically varies the relative familiarity of different stimuli enabling an examination of how neural activity changes as a function of experience. The data collected during this passive viewing task revealed that neurons in the MTL are generally not sensitive to the relative familiarity of a stimulus. In addition, when the specificity (i.e., which images a neuron was selective for) of individual neurons was analyzed, there was a significant dissociation between different medial temporal regions, with only neurons in TF, but not CA3 or the perirhinal cortex, altering their activity as stimuli became familiar. The implications of these findings are discussed in the context of how MTL structures process information during a passive viewing paradigm.
macaque; sparse; medial temporal lobe; passive viewing; tuning
The CA1 region of the hippocampus receives distinct patterns of afferent input to distal (near subiculum) and proximal (near CA2) zones. Specifically, distal CA1 receives a direct projection from cells in the lateral entorhinal cortex that are sensitive to objects, while proximal CA1 is innervated by cells in the medial entorhinal cortex that are responsive to space. This suggests neurons in different areas along the promixodistal axis of CA1 of the hippocampus will be functionally distinct. The current experiment investigated this possibility by monitoring behavior-induced cell activity across the CA1 axis using Arc mRNA imaging methods that compared adult and old rats in two conditions: exploration of the same environment containing the same objects twice (AA), or exploration of two different environments that contained identical objects (AB). The hypothesis was that CA1 place cells should show field remapping in the condition where environments were changed, but the extent of remapping was expected to differ between proximal and distal regions, and between age groups. In fact, neurons in the proximal region of CA1 in adult animals exhibited a greater degree of remapping than did distal CA1 cells when the environment changed, suggesting that cells receiving input from the medial entorhinal cortex are more sensitive to spatial context. In old rats, however, there were no differences in remapping across the proximodistal CA1 axis. Together these data suggest that distal and proximal CA1 may be functionally distinct and differentially vulnerable to normative aging processes.
Age-related cognitive and behavioral slowing may be caused by changes in the speed of neural signaling or by changes in the number of signaling steps necessary to achieve a given function. In the mammalian cortex, neural communication is organized by a 30–100 Hz “gamma” oscillation. There is a putative link between the gamma frequency and the speed of processing in a neural network: the dynamics of pyramidal neuron membrane time constants suggest that synaptic integration is framed by the gamma cycle, and pharmacological slowing of gamma also slows reaction times on behavioral tasks. The present experiments identify reductions in a robust 40–70 Hz gamma oscillation in the aged rat medial frontal cortex. The reductions were observed in the form of local field potentials (LFPs), later peaks in fast-spiking neuron autocorrelations, and delays in the spiking of inhibitory neurons following local excitatory signals. Gamma frequency did not vary with movement speed, but rats with slower gamma also moved more slowly. Gamma frequency age differences were not observed in hippocampus. Hippocampal CA1 fast-spiking neurons exhibited inter-spike intervals consistent with a fast (70–100 Hz) gamma frequency, a pattern maintained across theta phases and theta frequencies independent of fluctuations in the neurons’ average firing rates. We propose that an average lengthening of the cortical 15–25 ms gamma cycle is one factor contributing to age-related slowing, and that future attempts to offset cognitive declines will find a target in the response of fast-spiking inhibitory neurons to excitatory inputs.
One prominent component of aging is a defect in memory stabilization. To understand how the formation of enduring memories is altered in the aged brain, long-term markers of the biological events that may mediate memory consolidation were used to examine the activity dynamics of hippocampal circuits over extended intervals. The immediate-early gene Arc, which is implicated in both durable memory and synaptic plasticity, is expressed in the fascia dentata (FD) for long periods following behavioral experience. To test the hypothesis that aging alters long-term Arc transcription in the FD, a region critical for spatial memory and impaired with progressive age, young and aged rats explored a novel environment twice, separated by an 8 hour interval, and FD Arc transcription was assessed. Relative to young rats, (a) fewer granule cells in the aged FD transcribe Arc 8 hours after spatial exploration, and (b) this decrease is correlated with impaired spatial memory. These findings are consistent with behavioral evidence of age-related decline in hippocampal-dependent memory processing long after an event is to be remembered, and re-affirm the integral role of the FD in the neural circuits supporting durable memory.
Arg3.1; dentate gyrus; IEG; granule cell; consolidation; long-term memory
Adult-born neurons in the dentate gyrus (DG) can survive for long periods, are capable of integrating into neuronal networks, and are important for hippocampus-dependent learning. Neurogenesis is dramatically reduced during senescence, and it remains unknown whether those few neurons that are produced remain capable of network integration. The expression of Arc, a protein coupled to neuronal activity, was used to measure activity among granule cells that were labeled with BrdU four months earlier in young (9 months) and aged (25 months) Fischer344 rats. The results indicate that while fewer cells are generated in the senescent DG, those that survive are (a) more likely to respond to spatial processing by expressing Arc relative to the remainder of the granule cell population, and (b) equally responsive to spatial exploration as granule cells of the same age from young animals. These findings provide compelling evidence that newborn granule cells in the aged DG retain the capacity for participation in functional hippocampal networks.
Aging; Neurogenesis; Arc; Dentate Gyrus
The need to map regions of brain tissue that are much wider than the field of view of the microscope arises frequently. One common approach is to collect a series of overlapping partial views, and align them to synthesize a montage covering the entire region of interest. We present a method that advances this approach in multiple ways. Our method (1) produces a globally consistent joint registration of an unorganized collection of 3-D multi-channel images with or without stage micrometer data; (2) produces accurate registrations withstanding changes in scale, rotation, translation and shear by using a 3-D affine transformation model; (3) achieves complete automation, and does not require any parameter settings; (4) handles low and variable overlaps (5 – 15%) between adjacent images, minimizing the number of images required to cover a tissue region; (5) has the self-diagnostic ability to recognize registration failures instead of delivering incorrect results; (6) can handle a broad range of biological images by exploiting generic alignment cues from multiple fluorescence channels without requiring segmentation; and (7) is computationally efficient enough to run on desktop computers regardless of the number of images. The algorithm was tested with several tissue samples of at least 50 image tiles, involving over 5,000 image pairs. It correctly registered all image pairs with an overlap greater than 7%, correctly recognized all failures, and successfully joint-registered all images for all tissue samples studied. This algorithm is disseminated freely to the community as included with the FARSIGHT toolkit for microscopy (www.farsight-toolkit.org).
Montage Synthesis; Image Registration; 3-D Microscopy
Spatiotemporal and recognition memory are affected by aging in humans and macaque monkeys. To investigate whether these deficits are coupled with atrophy of memory-related brain regions, T1-weighted magnetic resonance images were acquired and volumes of the cerebrum, ventricles, prefrontal cortex (PFC), calcarine cortex, hippocampus, and striatum were quantified in young and aged rhesus monkeys. Subjects were tested on a spatiotemporal memory procedure (delayed response [DR]) that requires the integrity of the PFC and a medial temporal lobe-dependent recognition memory task (delayed nonmatching to sample [DNMS]). Region of interest analyses revealed that age inversely correlated with striatal, dorsolateral prefrontal cortex (dlPFC), and anterior cingulate cortex volumes. Hippocampal volume predicted acquisition of the DR task. Striatal volume correlated with DNMS acquisition, whereas total prefrontal gray matter, prefrontal white matter, and dlPFC volumes each predicted DNMS accuracy. A regional covariance analysis revealed that age-related volumetric changes could be captured in a distributed network that was coupled with declining performance across delays on the DNMS task. This volumetric analysis adds to growing evidence that cognitive aging in primates arises from region-specific morphometric alterations distributed across multiple memory-related brain systems, including subdivisions of the PFC.
age-related memory impairment; medial temporal lobe; MRI; prefrontal cortex; rhesus monkey
The hippocampal formation has been implicated in a growing number of disorders, from Alzheimer's disease and cognitive ageing to schizophrenia and depression. How can the hippocampal formation, a complex circuit that spans the temporal lobes, be involved in a range of such phenotypically diverse and mechanistically distinct disorders? Recent neuroimaging findings indicate that these disorders differentially target distinct subregions of the hippocampal circuit. In addition, some disorders are associated with hippocampal hypometabolism, whereas others show evidence of hypermetabolism. Interpreted in the context of the functional and molecular organization of the hippocampal circuit, these observations give rise to a unified pathophysiological framework of hippocampal dysfunction.
The gradual decline of cognitive ability with age, even in the absence of overt brain disease, is a growing problem. Although cognitive aging is a common and feared accompaniment of the aging process, its underlying mechanisms are not well understood and there are no highly effective means to prevent it. Additional research on cognitive aging is sorely needed, and methods that enable ready translation between human subjects and animal models stand to provide the most benefit. Here and in the six companion pieces in this special issue, we discuss a variety of challenges and opportunities for studying cognitive aging across species. We identify tests of associative memory, recognition memory, spatial and contextual memory, and working memory and executive function as cognitive domains that are age-sensitive and amenable to testing with parallel means in both humans and animal models. We summarize some of the important challenges in using animal models to test cognition. We describe unique opportunities to study cognitive aging in human subjects, such as those provided by recent large-scale initiatives to characterize cognition in large groups of subjects across the lifespan. Finally, we highlight some of the challenges of studying cognitive aging in human subjects.
cognition; aging; memory; human; animal models
An overview is provided of the simple single-cue delay and trace eyeblink conditioning paradigms as techniques to assess associative learning and memory in the aged. We highlight and focus this review on the optimization of the parameter space of eyeblink conditioning designs in the aged to avoid and control for potential confounds that may arise when studying aged mammals. The need to examine the contribution of non-associative factors that can contribute to performance outcomes is emphasized, and how age-related changes in the central nervous system as well as peripheral sensory factors can potentially bias the interpretation of the data in the aged is discussed. The way in which slight alterations of the parameter space in the delay and trace eyeblink conditioning paradigms can lead to delayed but intact conditioning, rather than impaired performance in aged animals is also discussed. Overall, the eyeblink conditioning paradigm, when optimized for the age of the animal in the study, is an elegantly simple technique for assessment of associative learning and memory. When design caveats described above are taken into account, this important type of memory, with its well-defined neural substrates, should definitely be included in cognitive assessment batteries for the aged.
associative learning; delay conditioning; trace conditioning; optimization; cognitive assessment battery
Analyses of complex behaviors across the lifespan of animals can reveal the brain regions that are impacted by the normal aging process, thereby, elucidating potential therapeutic targets. Recent data from rats, monkeys, and humans converge, all indicating that recognition memory and complex visual perception are impaired in advanced age. These cognitive processes are also disrupted in animals with lesions of the perirhinal cortex, indicating that the the functional integrity of this structure is disrupted in old age. This current review summarizes these data, and highlights current methodologies for assessing perirhinal cortex-dependent behaviors across the lifespan.
medial temporal lobe; monkey; perception; perirhinal cortex; rat
CA1 cells receive direct input from space-responsive cells in medial entorhinal cortex (MEC), such as grid cells, as well as more non-spatial cells in lateral entorhinal cortex (LEC). Because MEC projects preferentially to the proximal part of the CA1, bordering CA2, whereas LEC innervates only the distal part, bordering subiculum, we asked if spatial tuning is graded along the transverse axis of CA1. Tetrodes were implanted along the entire proximodistal axis of dorsal CA1 in rats. Data were recorded in cylinders large enough to elicit firing at more than one location in many neurons. Distal CA1 cells showed more dispersed firing and had a larger number of firing fields than proximal cells. Phase-locking of spikes to MEC theta oscillations was weaker in distal CA1 than in proximal CA1. The findings suggest that spatial firing in CA1 is organized transversally, with the strongest spatial modulation occurring in the MEC-associated proximal part.
Over the past half century, remarkable progress has been made in understanding the biological basis of memory and how it changes over the lifespan. An important conceptual advance during this period was the realization that normative cognitive trajectories can exist independently of dementing illness. In fact, mammals as different as rats and monkeys, who do not spontaneously develop Alzheimer’s disease, show memory impairments at advanced ages in similar domains as those observed in older humans. Thus, animal models have been particularly helpful in revealing brain mechanisms responsible for the cognitive changes that occur in aging. During these past decades, a number of empirical and technical advances enabled the discoveries that began to link age-related changes in brain function to behavior. The pace of innovation continues to accelerate today, resulting in an expanded window through which the secrets of the aging brain are being deciphered.
To rely on the anatomical organization of the hippocampal formation to understand how late-life diseases such as diabetes and stroke contribute to age-related cognitive decline.
Magnetic resonance imaging (MRI) was used to document brain infarcts and to generate high-resolution functional maps of the hippocampal formation in 240 community-based non-demented elders (mean age=79.7) who received a comprehensive medical evaluation. Sixty participants had type 2 diabetes mellitus while 74 had MRI-documented brain infarcts, and the first analysis was designed to pinpoint hippocampal subregions differentially linked to each disorder. Then, guided by the results, additional fMRI studies in aging rhesus monkeys and mice were used to test proposed mechanisms of dysfunction.
Although both diabetes and brain infarcts were associated with hippocampal dysfunction, each was linked to separate hippocampal subregions, suggesting distinct underlying mechanisms. The hippocampal subregion linked to diabetes implicated blood glucose as a pathogenic mechanism, a hypothesis confirmed by imaging aging rhesus monkeys and a mouse model of diabetes. The hippocampal subregion linked to infarcts suggested transient hypoperfusion as a pathogenic mechanism, a hypothesis provisionally confirmed by comparing anatomical patterns across subjects with infarcts in different vascular territories.
Taken together with previous findings, these results clarify how diseases of late-life differentially target the hippocampal formation, identify causes that contribute to age-related cognitive decline, and suggest specific interventions that can preserve cognitive health.
Previously, utilizing a series of genome-wide association, brain imaging and gene expression studies we implicated the KIBRA gene and the RhoA/ROCK pathway in hippocampal-mediated human memory. Here we show that peripheral administration of the ROCK inhibitor hydroxyfasudil improves spatial learning and working memory in the rodent model. This study supports the action of ROCK on learning and memory, suggests the potential value of ROCK inhibition for the promotion of cognition in humans and highlights the powerful potential of unbiased genome-wide association studies to inform potential novel uses for existing pharmaceuticals.
learning; memory; ROCK; fasudil; aging
This brief review will focus on a new hypothesis for the role of epigenetic mechanisms in aging-related disruptions of synaptic plasticity and memory. Epigenetics refers to a set of potentially self-perpetuating, covalent modifications of DNA and post-translational modifications of nuclear proteins that produce lasting alterations in chromatin structure. These mechanisms, in turn, result in alterations in specific patterns of gene expression. Aging-related memory decline is manifest prominently in declarative/episodic memory and working memory, memory modalities anatomically based largely in the hippocampus and prefrontal cortex, respectively. The neurobiological underpinnings of age-related memory deficits include aberrant changes in gene transcription that ultimately affect the ability of the aged brain to be “plastic”. The molecular mechanisms underlying these changes in gene transcription are not currently known, but recent work points toward a potential novel mechanism, dysregulation of epigenetic mechanisms. This has led us to hypothesize that dysregulation of epigenetic control mechanisms and aberrant epigenetic “marks” drive aging-related cognitive dysfunction. Here we focus on this theme, reviewing current knowledge concerning epigenetic molecular mechanisms, as well as recent results suggesting disruption of plasticity and memory formation during aging. Finally, several open questions will be discussed that we believe will fuel experimental discovery.
memory; hippocampus; histone; DNA methylation; epigenetics; gene transcription; aging; cognitive impairment
During aging, many experience a decline in cognitive function that includes memory loss. The encoding of long-term memories depends on new protein synthesis, and this is also reduced during aging. Thus, it is possible that changes in the regulation of protein synthesis contribute to the memory impairments observed in older animals. Several lines of evidence support this hypothesis. For instance, protein synthesis is required for a longer period following learning to establish long-term memory in aged rodents. Also, under some conditions, synaptic activity or pharmacological activation can induce de novo protein synthesis and lasting changes in synaptic transmission in aged, but not young, rodents; the opposite results can be observed in other conditions. These changes in plasticity likely play a role in manifesting the altered place field properties observed in awake and behaving aged rats. The collective evidence suggests a link between memory loss and the regulation of protein synthesis in senescence. In fact, pharmaceuticals that target the signaling pathways required for induction of protein synthesis have improved memory, synaptic plasticity, and place cell properties in aged animals. We suggest that a better understanding of the mechanisms that lead to different protein expression patterns in the neural circuits that change as a function of age will enable the development of more effective therapeutic treatments for memory loss.
protein synthesis; translation; transcription; aging; hippocampus; memory; plasticity; place cells
Place fields of hippocampal pyramidal cells expand asymmetrically when adult rats repeatedly follow the same route. This behaviorally-induced expression of neuronal plasticity utilizes an NMDAR-dependent, LTP-like mechanism and could be used by hippocampal networks to store information. Aged spatial memory-impaired rats exhibit defective experience-dependent place field expansion plasticity. One possible explanation for this aged-associated deficit is alterations in glutamatergic function. In fact, both NMDAR- and AMPAR-mediated field excitatory postsynaptic potentials in CA1 decrease with aging. The current study investigated whether modulation of either AMPA or NDMA receptor activity could restore this experience-dependent plasticity by prolonging AMPAR activity with the ampakine CX516, and modulating the NMDAR with the noncompetitive antagonist memantine. The spatial firing characteristics of multiple CA1 pyramidal cells were monitored under both treatment conditions as aged rats repeatedly traversed a circular track. Compared to the saline baseline condition, acute administration of memantine but not CX516, reinstated experience-dependent place field expansion. Taken together, these data suggest that pharmacological manipulation of the NMDAR can improve the function of hippocampal networks critical to optimal cognition in aging.
aging; CA1; hippocampus; place cell; theta phase precession
Studies demonstrating recognition deficits with aging often use tasks in which subjects have an incentive to correctly encode or retrieve the experimental stimuli. In contrast to these tasks, which may engage strategic encoding and retrieval processes, the visual paired comparison (VPC) task measures spontaneous eye movements made toward a novel as compared with familiar stimulus. In the present study, seven rhesus macaques aged six to thirty years exhibited a dramatic age-dependent decline in preference for a novel image compared with one presented seconds earlier. The age effect could not be accounted for by memory deficits alone, since it was present even when familiarization preceded test by one second. It also could not be explained by an encoding deficit, since the effect persisted with increased familiarity of the sample stimulus. Reduced novelty preference did correlate with eye movement variables, including reaction time distributions and saccade frequency. At long delay intervals (24 or 48 hours) aging was paradoxically associated with increased novelty preference. Several explanations for the age effect are considered, including the possible role of dopamine.
aging; novelty; saccades; recognition memory; attention
The hippocampus is thought to coordinate memory consolidation by reactivating traces from behavioral experience when the brain is not actively processing new input. In fact, during slow-wave sleep the patterns of CA1 pyramidal cell ensemble activity correlations are reactivated in both young and aged rats. In addition to correlated activity patterns, repetitive track running also creates a recurring sequence of pyramidal cell activity. The present study compared CA1 sequence activity pattern replay in young and old animals during rest periods following behavior. While the young rats exhibited significant sequence reactivation, it was markedly impaired in the aged animals. When the spatial memory scores of all animals were compared to the degree of sequence reactivation, there was a significant correlation. The novel finding that weak replay of temporal patterns has behavioral consequences, strengthens the idea that reactivation processes are integral to memory consolidation.
aging; attractor dynamics; CA1; memory consolidation; place field; neural ensemble
Simultaneous imaging of multiple cellular components is of tremendous importance in the study of complex biological systems, but the inability to use probes with similar emission spectra and the time consuming nature of collecting images on a confocal microscope are prohibitive. Hyperspectral imaging technology, originally developed for remote sensing applications, has been adapted to measure multiple genes in complex biological tissues. A spectral imaging microscope was used to acquire overlapping fluorescence emissions from specific mRNAs in brain tissue by scanning the samples using a single fluorescence excitation wavelength. The underlying component spectra obtained from the samples are then separated into their respective spectral signatures using multivariate analyses, enabling the simultaneous quantitative measurement of multiple genes either at regional or cellular levels.
hyperspectral imaging; immediate early genes; fluorescence imaging; pushbroom line-imaging; multivariate image analysis; spectral unmixing
Introduction: Patients with complex medical care needs often embark on multiple care transitions over an extended period of time. As these patients or their caregivers often become the chief source of communication for complex medical situations, each transition can create an opportunity for health care errors. Combining the efforts of the established departments of Chronic Care Coordination (CCC), Clinical Pharmacy Call Center (CPCC), and Continuing Care, Kaiser Permanente Colorado created programs to further safe care transitions.
Methods: Two key goals for safe care transitions were established: 1) reductions in medication errors and 2) increased follow-up with care plans. To achieve these goals, communication plans targeted at medication reconciliation, patient education, and coordination of outpatient recommendations were established. Expected outcomes included reductions in medication errors, decreased Emergency Department and hospital admissions, decreased readmissions, and increased outpatient follow-up and medication compliance.
Results: A review of medication-reconciliation records for intervention patients indicated that >90% of all discharge summaries contained at least one potential drug-related problem including duplicative drugs, omitted therapy, and medication contraindications. After skilled nursing facility discharge, patients who were transitioned by CPCC clinical pharmacists were: 1) 78% less likely to die; 2) 29% less likely to need an Emergency Department visit; and 3) 17% more likely to follow up with primary physicians and clinicians than were patients in the usual care group. Health care cost savings for patients seen by the CCC program demonstrated, conservatively, an annualized per patient savings of $5276. For 763 patients enrolled in 2003, this amounts to an estimated, annualized savings of $4,025,588.
Conclusions: Patients are becoming more informed and involved in their care, but they require ongoing education and coaching to become effective advocates for themselves. Identification of unintended medication discrepancies and potential drug-related problems and increased follow-up during care transitions can improve patient safety and quality of care while saving health care resources.