Thousands of hippocampal neurons are born in adulthood, suggesting that new cells could be important for hippocampal function. To determine whether hippocampus-dependent learning affects adult-generated neurons, we examined the fate of new cells labeled with the thymidine analog bromodeoxyuridine following specific behavioral tasks. Here we report that the number of adult-generated neurons doubles in the rat dentate gyrus in response to training on associative learning tasks that require the hippocampus. In contrast, training on associative learning tasks that do not require the hippocampus did not alter the number of new cells. These findings indicate that adult-generated hippocampal neurons are specifically affected by, and potentially involved in, associative memory formation.

The vertebrate brain continues to produce new neurons throughout life. In the rat hippocampus, several thousand are produced each day, many of which die within weeks. Associative learning can enhance their survival; however, until now it was unknown whether new neurons are involved in memory formation. Here we show that a substantial reduction in the number of newly generated neurons in the adult rat impairs hippocampal-dependent trace conditioning, a task in which an animal must associate stimuli that are separated in time. A similar reduction did not affect learning when the same stimuli are not separated in time, a task that is hippocampal-independent. The reduction in neurogenesis did not induce death of mature hippocampal neurons or permanently alter neurophysiological properties of the CA1 region, such as long-term potentiation. Moreover, recovery of cell production was associated with the ability to acquire trace memories. These results indicate that newly generated neurons in the adult are not only affected by the formation of a hippocampal-dependent memory, but also participate in it.

ABSTRACT:The hippocampal formation generates new neurons throughout adulthood. Recent studies indicate that these cells possess the morphology and physiological properties of more established neurons. However, the function of adult generated neurons is still a matter of debate. We previously demonstrated that certain forms of associative learning can enhance the survival of new neurons and a reduction in neurogenesis coincides with impaired learning of the hippocampal-dependent task of trace eyeblink conditioning. Using the toxin methylazoxymethanol acetate (MAM) for proliferating cells, we tested whether reduction of neurogenesis affected learning and performance associated with different hippocampal dependent tasks: spatial navigation learning in a Morris water maze, fear responses to context and an explicit cue after training with a trace fear paradigm. We also examined exploratory behavior in an elevated plus maze. Rats were injected with MAM (7 mg/kg) or saline for 14 days, concurrent with BrdU, to label new neurons on days 10, 12, and 14. After treatment, groups of rats were tested in the various tasks. A significant reduction in new neurons in the adult hippocampus was associated with impaired performance in some tasks, but not with others. Specifically, treatment with the antimitotic agent reduced the amount of fear acquired after exposure to a trace fear conditioning paradigm but did not affect contextual fear conditioning or spatial navigation learning in the Morris water maze. Nor did MAM treatment affect exploration in the elevated plus maze. These results combined with previous ones suggest that neurogenesis may be associated with the formation of some but not all types of hippocampal-dependent memories.

During the past several years, evidence has accumulated suggesting a relationship between newly born cells in the hippocampus and various types of learning and memory. However, most of the evidence is correlational and some of it does not agree. This review discusses both sides of this issue, considering the effects of learning on the production of new neurons in the dentate gyrus and the question of whether newly born cells participate in learning and memory.

Dendritic spines are postsynaptic sites of excitatory input in the mammalian nervous system. Despite much information about their structure, their functional significance remains unknown. It has been reported that females in proestrus, when estrogen levels are elevated, have a greater density of apical dendritic spines on pyramidal neurons in area CA1 of the hippocampus than females in other stages of estrous ). Here we replicate these findings and in addition, show that females in proestrus have a greater density of spines in area CA1 of the hippocampus than males. Moreover, this sex difference in spine density is affected in opposite directions by stressful experience. In response to one acute stressful event of intermittent tailshocks, spine density was enhanced in the male hippocampus but reduced in the female hippocampus. The decrease in the female was observed for those that were stressed during diestrus 2 and perfused 24 hr later during proestrus. The opposing effects of stress were not evident immediately after the stressor but rather occurred within 24 hr and were evident on apical and to a lesser extent on basal dendrites of pyramidal cells in area CA1. Neither sex nor stress affected spine density on pyramidal neurons in somatosensory cortex. Sex differences in hippocampal spine density correlated with sex hormones, estradiol and testosterone, whereas stress effects on spine density were not directly associated with differences in the stress hormones, glucocorticoids. In summary, males and females have different levels of dendritic spine density in the hippocampus under unstressed conditions, and their neuronal anatomy can respond in opposite directions to the same stressful event.

Synaptic strengthening induced by brain-derived neurotrophic factor (BDNF) is associated with learning and is coupled to transcriptional activation. However, identification of the spectrum of genes associated with BDNF-induced synaptic plasticity and the correlation of expression with learning paradigms in vivo has not yet been studied. Transcriptional analysis of BDNF-induced synaptic strengthening in cultured hippocampal neurons revealed increased expression of the immediate early genes (IEGs), c-fos, early growth response gene 1 (EGR1), activity-regulated cytoskeletal-associated protein (Arc) at 20 min, and the secreted peptide VGF (non-acronymic) protein precursor at 3 hr. The induced genes served as prototypes to decipher mechanisms of both BDNF-induced transcription and plasticity. BDNF-mediated gene expression was tyrosine kinase B and mitogen-activated protein kinase-dependent, as demonstrated by pharmacological studies. Single-cell transcriptional analysis of Arc after whole-cell patch-clamp recordings indicated that increased gene expression correlated with enhancement of synaptic transmission by BDNF. Increased expression in vitro predicted elevations in vivo: VGF and the IEGs increased after trace eyeblink conditioning, a hippocampal-dependent learning paradigm. VGF protein was also upregulated by BDNF treatment and was expressed in a punctate manner in dissociated hippocampal neurons. Collectively, these findings suggested that the VGF neuropeptides may regulate synaptic function. We found a novel function for VGF by applying VGF peptides to neurons. C-terminal VGF peptides acutely increased synaptic charge in a dose-dependent manner, whereas N-terminal peptide had no effect. These observations indicate that gene profiling in vitro can reveal new mechanisms of synaptic strengthening associated with learning and memory.

A group of rats was trained to escape low-intensity shock in a shuttle-box test, while another group of yoked controls could not escape but was exposed to the same amount and regime of shock. After 1 week of training, long-term potentiation (LTP) was measured in vitro in hippocampal slices. Exposure to uncontrollable shock massively impaired LTP relative to exposure to the same amount and regime of controllable shock. These results provide evidence that controllability modulates plasticity at the cellular-neuronal level.

Dendritic spines are sources of synaptic contact that can be altered by experience and, as such, may be involved in memories for that experience. Here we tested whether the acquisition of new memories is associated with changes in the density of dendritic spines. Adult male rats were trained using the trace eyeblink conditioning paradigm, an associative learning task that requires the hippocampus for acquisition. Additional groups were exposed to the same number of stimuli presented in an explicitly unpaired manner or were naive. Twenty-four hours later, the density of dendritic spines was measured using Golgi impregnation. Trace conditioning was associated with an increase in the density of dendritic spines on the pyramidal cells of area CA1 of the hippocampus, an effect that was prevented by blocking acquisition of the learned response with a competitive NMDA receptor antagonist. Training with delay conditioning, a similar task that does not require the hippocampus, also produced an increase in spine density. The learning-induced increase in dendritic spine density was specific to basal dendrites of pyramidal cells in the CA1 region of the hippocampus. Changes did not occur on their apical dendrites or on cells in the dentate gyrus or somatosensory cortex. These results suggest that the formation and expression of associative memories increase the availability of dendritic spines and the potential for synaptic contact.