Published by Wiley
Online ISSN: 1098-1063
Print ISSN: 1050-9631
Carbachol, a muscarinic receptor agonist, produced three distinct spontaneous oscillations in the CA3 region of rat hippocampal slices. Carbachol concentrations in the 4-13 microM range produced regular synchronized CA3 discharges at 0.5-2 Hz (carbachol-delta). Higher concentrations (13-60 microM) produced short episodes of 5-10 Hz (carbachol-theta) oscillations separated by nonsynchronous activity. Concentrations of carbachol ranging from 8-25 microM also produced irregular episodes of high-frequency discharges (carbachol-gamma, 35-70 Hz), in isolation or mixed with carbachol-theta and carbachol-delta. At carbachol concentrations sufficient to induce carbachol-theta, low concentrations of APV reversibly transformed carbachol-theta into carbachol-delta. Higher concentrations of D,L-2-amino-5-phosphonopentanoic acid (APV) reversibly and completely blocked carbachol-theta. A systematic study of the effects of carbachol shows that the frequency of spontaneous oscillations depended nonlinearly on the level of muscarinic activation. Field and intracellular recordings from CA1 and CA3 pyramidal cells and interneurons during carbachol-induced rhythms revealed that the hippocampal circuitry preserved in the slice was capable of spontaneous activity over the range of frequencies observed in vivo and suggests that the presence of these rhythms could be under neuromodulatory control.
Effect of acute administration of JWH-081 on ERK, CaMKIV, and CREB phosphorylation levels in the hippocampus. (A) Western blot analyses were performed to determine the levels of pERK1/2 and ERK1/2 in hippocampal total extracts. β-actin was used as a loading control. Representative blots are shown for the hippocampal total extracts (n = 6 mice/group). (B) Hippocampal nuclear extracts from WT and KO mice treated with or without JWH-081 (1.25 mg/kg) for 30 or 60 min were processed for Western blot to analyze the levels of pCaMKIV, pCREB, CaMKIV and CREB. β-actin was used as a loading control. ***P < 0.001; **P < 0.01. (C) Hippocampal nuclear extracts from vehicle and SR pretreated C57BL/6J mice treated with or without JWH-081 (1.25 mg/kg) for 30 min were processed for Western blot to analyze the levels of pCaMKIV (# and $ P < 0.05), pCREB (# and $ P < 0.01), CaMKIV and CREB. β-actin was used as a loading control. Representative blots are shown for the hippocampal nuclear extracts (n = 6 mice/group). One-way ANOVA with Bonferroni's post hoc test was used for statistical analysis. The error bars represent SEM. ***P < 0.001 vs. vehicle (V); # vs. vehicle; $ vs. JWH-081.
JWH-081 inhibits LTP in hippocampal slices from adult CB1 receptor WT but not KO mice. (A) A schematic diagram showing the stimulating and recording electrode positions in the CA1 region of the hippocampus. (B) A summary graph showing the field input/output relationships in slices after treating WT slices with vehicle or JWH-081. (C) A time course of the averages of the fEPSP slopes in slices after treating WT slices with vehicle or JWH-081. (D) A summary graph showing the field input/output relationships in slices after treating KO slices with vehicle or JWH-081. (E) A time course of the averages of the fEPSP slopes in slices after treating KO slices with vehicle or JWH-081.The fEPSP slopes were normalized to the average value 10 min before stimulation in each experiment. Arrows denote the time of tetanic stimulation (4 pulses at 100 Hz, with bursts repeated at 5 Hz, each tetanus included three 10-burst trains separated by 15 s). For JWH-081 treatment experiments, after 10 min of baseline recording, hippocampal slices were perfused with JWH-081 (1.0 μM in DMSO) or vehicle (0.001% DMSO) for 30 min before inducing (solid line) LTP with tetanic stimulation of the Schaeffer collateral pathway. (F) A combined plot of the averages of the fEPSP slopes for WT and KO slices with and without JWH-081 at several time points. Each point is presented as the mean ± SEM (n= 5 mice/group; 10 slices/group). One-way ANOVA with Bonferroni's post hoc tests; ***P < 0.001.
Acute administration of JWH-081 impairs spontaneous alternation performance in adult mice. (A) Spatial working memory of CB1 receptor WT and KO mice treated with or without JWH-081 was tested using spontaneous alternation performance in the Y-maze. Note that JWH-081 treated WT mice perform poorly (well below 50% chance levels) compared with WT mice treated with vehicle (***P < 0.001). Overall, KO mice treated with either vehicle or JWH-081 show significantly enhanced levels of alternation performance (#P < 0.05 vs. CB1WT + V; @P < 0.05 vs. CB1WT + V; $P < 0.05 vs. CB1WT + JWH-081) when compared with WT mice with or without JWH-081 treatment. (B) The number of arm entries was not different between WT and KO mice (P > 0.05) but JWH-081 treatment enhanced the number of arm entries in both WT (***P < 0.001 vs. CB1WT + V) and KO mice (***P < 0.001 vs. CB1KO + V). (C) The time spent in each arm was not different between WT and KO mice (P > 0.05) with or without JWH-081 treatment (P > 0.05 vs. CB1WT + V). Each point is the mean ± SEM (n = 8 mice/group). One-way ANOVA with Bonferroni's post hoc test.
Recently, synthetic cannabinoids have been sprayed onto plant material, which is subsequently packaged and sold as "Spice" or "K2" to mimic the effects of marijuana. A recent report identified several synthetic additives in samples of "Spice/K2"including JWH-081, a synthetic ligand for the cannabinoid receptor 1 (CB1). The deleterious effects of JWH-081 on brain function are not known, particularly on CB1 signaling, synaptic plasticity, learning and memory. Here, we evaluated the effects of JWH-081 on pCaMKIVpCREB and pERK1/2 signaling events followed by long-term potentiation (LTP), hippocampal-dependent learning and memory tasks using CB1 receptor wild type (WT) and knockout (KO) mice. Acute administration of JWH-081 impaired CaMKIV phosphorylation in a dose-dependent manner, whereas inhibition of CREB phosphorylation in CB1 receptor WT mice was observed only at higher dose of JWH-081 (1.25 mg/kg). JWH-081 at higher dose impaired CaMKIV and CREB phosphorylation in a time -dependent manner in CB1 receptor WT mice but not in KO mice and failed to alter ERK1/2 phosphorylation. In addition, SR treated or CB1 receptor KO mice have a lower pCaMKIV/CaMKIV ratio and higher pCREB/CREB ratio compared to vehicle or WT littermates. In hippocampal slices, JWH-081 impaired LTP in CB1 receptor WT but not in KO littermates. Furthermore, JWH-081 at higher dose impaired object recognition, spontaneous alternation and spatial memory on the Y-maze in CB1 receptor WT mice but not in KO mice. Collectively our findings suggest that deleterious effects of JWH-081 on hippocampal function involves CB1 receptor mediated impairments in CaMKIV and CREB phosphorylationLTP, learning and memory in mice.
In this study, we employed 1α-hydroxylase knockout (1α-(OH)ase(-/-) ) mice to investigate the influence of 1,25-dihydroxy vitamin D(3) (1,25-(OH)(2) D(3) ) deficiency on the adult neurogenesis in the hippocampal dentate gyrus (DG). The numbers of both 24-hr-old BrdU(+) cells and proliferating cell nuclear antigen positive cells in 8-week-old 1α-(OH)ase(-/-) mice increased approximately twofold compared with wild-type littermates. In contrast, the numbers of 7- and 28-day-old BrdU(+) cells in 1α-(OH)ase(-/-) mice decreased by 50% compared with wild-type mice, while the proportion of BrdU(+) /NeuN(+) cells in BrdU(+) population showed no difference between 1α-(OH)ase(-/-) and wild-type mice. Apoptotic cells in the subgranular zone (SGZ) of DG markedly increased in 1α-(OH)ase(-/-) mice. Replenishment of 1,25-(OH)(2) D(3) , but not correction of serum calcium and phosphorus levels, completely prevented changes in the neurogenesis in 1α-(OH)ase(-/-) mice. The absence of 1,25-(OH)(2) D(3) led to an increase in the expression of L-type voltage-gated calcium channel (L-VGCC) and a decrease in the nerve growth factor (NGF) mRNA level. Treatment with the L-VGCC inhibitor nifedipine blocked the increased cell proliferations by 1,25-(OH)(2) D(3) deficiency. Administration of NGF significantly attenuated the loss of newborn neurons in 1α-(OH)ase(-/-) mice.
Long-lasting effects of a single exposure to IMO on Morris water maze learning. Mean and SEM are represented. There were no differences between groups in visible platform training in the water maze regarding latency (although all groups showed a good learning curve) (A) or average speed to find the visible platform (B). Invisible platform training showed no differences between groups in latency (C) or average speed to find the platform (D), although latency decreased over the days. Percent time spent in the target area (T) and in the opposite area (O) was evaluated in three different probe trials by circular areas analysis (E): probe trials 1 and 2 were considered a measure of ST memory, whereas the 24-h probe trial measured LT memory. No differences between groups were observed in probe trials 1 and 2, although, there were differences between time spent in T and O areas, indicating preference for the platform area in all four groups: probe trial 1 (P1), +++P < 0.001; probe trial 2 (P2), ++P < 0.01. In the 24-h probe trial, only control groups (Control LL3 and Control LL9) showed preference for the T area (+++P < 0.001 percent time in T as compared to O area), whereas no differences between the percent time spent in both areas were found in IMO groups (IMO LL3 and IMO LL9). Moreover, there were differences in the percent time spent in the target area in control versus IMO groups (**P < 0.01).
Effects of 7,8-DHF administration on the HPA activity under resting conditions or in response to IMO. Mean and SEM of plasma ACTH (A) and corticosterone (B) are represented. Rats received vehicle (Veh) or 7,8-DHF 2 h before stress. Half of the animals of each treatment were exposed to 2-h IMO and sampled, whereas the remaining (controls) were handled. ACTH and corticosterone were evaluated: immediately after stress and at 45 and 90-min poststress (R45 and R90). No effect of the drug was found either in control or IMO groups. Significant increases in ACTH and corticosterone were observed just after IMO and at R45 (***P < 0.001 vs. respective sampling time in controls).
7,8-DHF, given before or after IMO, blocks long-term stress-induced impairment of spatial memory. Mean and SEM of percent time spent in the circular target area (T) and in the circular opposite area (O) are represented. In Experiment 2, rats were given vehicle (Veh) or 7,8-DHF and 2 h later exposed to 2 h IMO. Spatial-learning procedure started 3 days after IMO. The results can be seen in panel A: probe trials 1 and 2 reflect ST memory and 24-h probe trial reflects LT memory. No differences between groups are observed in ST memory, and all of them learned the task as revealed by statistically differences between percent time in T versus O areas (+++P < 0.001 in the two probe trials). In the 24-h probe trial, all the groups except Veh–IMO showed preference for T as compared to O (++P < 0.01; +P < 0.05). In addition, there were significant differences between the Veh–IMO and the other three groups in the time spent in T and O areas (*P at least <0.05 vs. all other groups within the same area). In Experiment 3 (panel B), rats remained unstressed (controls) or were exposed to 2 h IMO and 8 h after the termination of IMO received vehicle or 7,8-DHF (IMO–Veh and IMO–7,8-DHF). Three days later, spatial-learning procedure started. Again, no differences between groups were observed in ST memory (probe trials 1 and 2), and all rats spent more time in T versus O areas (+++P < 0.001). In contrast, group differences were found in LT memory: IMO–Veh group showed no preference for T as compared to O areas, whereas the other two groups showed the expected difference (+++P < 0.001). Finally, the IMO–Veh group spent less time than the other two groups in the T area (**P < 0.01).
Post-traumatic stress disorder (PTSD) patients show cognitive deficits, but it is unclear whether these are a consequence of the pathology or a pre-existing factor of vulnerability to PTSD. Animal models may help to demonstrate whether or not exposure to certain stressors can actually induce long-lasting (LL; days) impairment of hippocampus-dependent memory tasks and to characterize neurobiological mechanisms. Adult male rats were exposed to 2-h immobilization on boards (IMO), a severe stressor, and spatial learning in the Morris water maze (MWM) was studied days later. Exposure to IMO did not modify learning or short-term memory in the MWM when learning started 3 or 9 days after IMO, but stressed rats did show impaired long-term memory at both times, in accordance with the severity of the stressor. New treatments to prevent PTSD symptoms are needed. Thus, considering the potential protective role of brain-derived neurotrophic factor (BDNF) on hippocampal function, 7,8-dihydroxyflavone (7,8-DHF), a recently characterized agonist of the BDNF receptor TrkB, was given before or after IMO in additional experiments. Again, exposure to IMO resulted in LL deficit in long-term memory, and such impairment was prevented by the administration of 7,8-DHF either 2 h prior IMO or 8 h after the termination of IMO. The finding that IMO-induced impairment of spatial memory was prevented by pharmacological potentiation of TrkB pathway with 7,8-DHF even when the drug was given 8 h after IMO suggests that IMO-induced impairment is likely to be a LL process that is strongly dependent on the integrity of the BDNF-TrkB system and is susceptible to poststress therapeutic interventions. 7,8-DHF may represent a new therapeutic approach for early treatment of subjects who have suffered traumatic experiences.
The anatomical distribution of sensory-evoked activity recorded from the hippocampal long-axis can shift depending on prior experience. In accordance with Marr's computational model of hippocampal function, CA3 NMDA receptors have been hypothesized to mediate this experience-dependent shift in hippocampal activity. Here we tested this hypothesis by investigating genetically-modified mice in which CA3 NMDA receptors are selectively knocked-out (CA3-NR1 KO). First, we were required to develop an fMRI protocol that can record sensory-evoked activity from the mouse hippocampal long-axis. This goal was achieved in part by using a dedicated mouse scanner to image odor-evoked activity, and by using non-EPI (echo planer imaging) pulse sequences. As in humans, odors were found to evoke a ventral-predominant activation pattern in the mouse hippocampus. More importantly, odor-evoked activity shifted in an experience-dependent manner. Finally, we found that the experience-dependent shift in hippocampal long-axis activity is blocked in CA3-NR1 knock-out mice. These findings establish a cellular mechanism for the plasticity imaged in the hippocampal long-axis, suggesting how experience-dependent modifications of hippocampal activity can contribute to its mnemonic function.
Summary of studies examining RA of contextual fear from our laboratory. A: From Kim and Fanselow (1992). Rats were given 10 tone-shock pairings and then given electrolytic lesions of the DH 1, 7, 14, or 28 days after training. After recovery, contextual freezing (% time, mean SEM) was assessed. A substantial deficit was observed when lesions were made 1 day, but not 28 days, after training. B: From Maren et al. (1997). Rats were given three tone-shock pairings and then given excitotoxic NMDA lesions of the DH 1, 28, or 100 days after training. After recovery, contextual freezing (peak % time, mean SEM) was assessed. A severe deficit was observed in rats that received lesions 1 day after training, but only a very mild deficit
Remote context preexposure protects rats from hippocampal lesion-induced RA of recent contextual fear. Rats were preexposed to one context for 60 min (without any shock or tone) and 35 days later were given Pavlovian fear conditioning in the same context and a highly distinct novel context. One to 2 days after conditioning, the animals received either sham or electrolytic DH lesions. Plotted here are the results of two 6-min context fear tests taken after recovery (% time freezing, mean SEM). DH lesions produced a severe RA of fear to the not-preexposed context, but only a mild, nonsignificant impairment of fear to the preexposed context.
Dorsal hippocampal (DH) lesions produce a severe deficit in recently, but not remotely, acquired contextual fear without impairing memory of discrete training stimuli, i.e., DH lesions produce an anterograde and time-limited retrograde amnesia specific to contextual memory. These data are consistent with the standard model which posits temporary involvement of the hippocampus in recent memory maintenance. However, three recent controversies apparently weaken the case for a selective mnemonic role for the hippocampus in contextual fear. First, although retrograde amnesia (from posttraining lesions) is severe, anterograde amnesia (from pretraining lesions) may be mild or nonexistent. Second, a performance, rather than mnemonic, account of contextual freezing deficits in hippocampal-lesioned animals has been offered. Third, damage to the entire hippocampus, including the ventral hippocampus, can produce a dramatic and temporally stable disruption of context and tone fear. These data are reviewed and explanations are offered as to why they do not necessarily challenge the standard model of hippocampal memory function in contextual fear. Finally, a more complete description of the hippocampus' proposed role in contextual fear is offered, along with new data supporting this view. In summary, the data support a specific mnemonic role for the DH in the acquisition and consolidation of contextual representations.
Neurogranin/RC3 is a protein that binds calmodulin and serves as a substrate for protein kinase C. Neuronally distributed in the hippocampus and forebrain, neurogranin is highly expressed in dendritic spines of hippocampal pyramidal cells, implicating this protein in long-term potentiation and in learning and memory processes. Null mutation of the neurogranin gene Ng generated viable knockout mice for analysis of the behavioral phenotype resulting from the absence of neurogranin protein. Ng -/- mice were normal on measures of general health, neurological reflexes, sensory abilities, and motor functions, as compared to wild type littermate controls. On the Morris water task, Ng -/- mice failed to reach acquisition criterion on the hidden platform test and did not show selective search on the probe trial. In the Barnes circular maze, another test for spatial navigation learning, Ng -/- mice showed impairments on some components of transfer, but normal performance on time spent around the target hole. Abnormal and idiosyncratic behaviors were detected, that appeared to represent an anxiogenic phenotype in Ng -/- mice, as measured in the light<-->dark exploration test and the open field center time parameter. These findings of apparent deficits in spatial learning and anxiety-like tendencies in Ng -/- support a role for neurogranin in the hippocampally-mediated interaction between stress and performance.
Differences between isogenic mouse strains in cellular expression of the neuronal nicotinic acetylcholine (ACh) receptor subunit alpha 4 (nAChR alpha 4) by the dorsal hippocampus are well known. To investigate further the genetic basis of these variations, expression of the nAChR alpha 4 subunit was measured in congenic mouse lines derived from two strains exhibiting notable divergence in the expression of this subunit: C3H and C57BL/6. Congenic lines carrying reciprocally introgressed regions (quantitative trait loci; QTL) from chromosomes 4, 5, and 12 each retained the phenotype most closely associated with the parental strain. However, in congenic lines harboring the reciprocal transfer of a chromosome 11 QTL, a characteristic difference in the ratio of interneurons versus astrocytes expressing nAChR alpha 4 in the CA1 region is reversed relative to the parental strain. These finding suggest that this chromosomal segment harbors genes that regulate strain distinct hippocampal morphology that is revealed by nAChR alpha 4 expression.
Hippocampal neurogenesis declines substantially in chronic temporal lobe epilepsy (TLE). However, it is unclear whether this decline is linked to altered production of new cells and/or diminished survival and neuronal fate-choice decision of newly born cells. We quantified different components of hippocampal neurogenesis in rats exhibiting chronic TLE. Through intraperitoneal administration of 5'-bromodeoxyuridine (BrdU) for 12 days, we measured numbers of newly born cells in the subgranular zone-granule cell layer (SGZ-GCL) at 24 h and 2.5 months post-BrdU administration. Furthermore, the differentiation of newly added cells into neurons and glia was quantified via dual immunofluorescence for BrdU and various markers of neurons and glia. Addition of new cells to the SGZ-GCL over 12 days was comparable between the chronically epileptic hippocampus and the age-matched intact hippocampus. Furthermore, comparison of BrdU+ cells measured at 24 h and 2.5 months post-BrdU administration revealed similar survival of newly born cells between the two groups. However, only 4-5% of newly born cells (i.e., BrdU+ cells) differentiated into neurons in the chronically epileptic hippocampus, in comparison to 73-80% of such cells exhibiting neuronal differentiation in the intact hippocampus. Moreover, differentiation of newly born cells into S-100beta+ astrocytes or NG2+ oligodendrocyte progenitors increased to approximately 79% in the chronically epileptic hippocampus from approximately 25% observed in the intact hippocampus. Interestingly, the extent of proliferation of astrocytes and microglia (identified through Ki-67 and S-100beta and Ki-67 and OX-42 dual immunofluorescence) in the SGZ-GCL was similar between the chronically epileptic hippocampus and the age-matched intact hippocampus, implying that the proliferation of neural stem/progenitor cells in the SGZ-GCL of the chronically epileptic hippocampus was not obscured by an increased division of glia. Thus, severely diminished DG neurogenesis in chronic TLE is not associated with either decreased production of new cells or reduced survival of newly born cells in the SGZ-GCL. Rather, it is linked to a dramatic decline in the neuronal fate-choice decision of newly generated cells. Overall, the differentiation of newly born cells turns mainly into glia with chronic TLE from predominantly neuronal differentiation seen in control conditions.
To explore the role of adult hippocampal neurogenesis in novelty processing, we assessed novel object recognition (NOR) in mice after neurogenesis was arrested using focal x-irradiation of the hippocampus, or a reversible, genetic method in which glial fibrillary acidic protein-positive neural progenitor cells are ablated with ganciclovir. Arresting neurogenesis did not alter general activity or object investigation during four exposures with two constant objects. However, when a novel object replaced a constant object, mice with neurogenesis arrested by either ablation method showed increased exploration of the novel object when compared with control mice. The increased novel object exploration did not manifest until 4-6 weeks after x-irradiation or 6 weeks following a genetic ablation, indicating that exploration of the novel object is increased specifically by the elimination of 4- to 6-week-old adult born neurons. The increased novel object exploration was also observed in older mice, which exhibited a marked reduction in neurogenesis relative to young mice. Mice with neurogenesis arrested by either ablation method were also impaired in one-trial contextual fear conditioning (CFC) at 6 weeks but not at 4 weeks following ablation, further supporting the idea that 4- to 6-week-old adult born neurons are necessary for specific forms of hippocampal-dependent learning, and suggesting that the NOR and CFC effects have a common underlying mechanism. These data suggest that the transient enhancement of plasticity observed in young adult-born neurons contributes to cognitive functions.
Károly Schaffer (1864-1939) was a Hungarian neurologist who distinguished himself through original discoveries in human neuropathology. At the beginning of his scientific carrier, he described the cellular and fiber structure of the hippocampus, earning him a high reputation in neuroscience. Schaffer (1892) described the so-called "collateral fiber system" that connects the CA3 and CA1 regions of the hippocampus, known today as Schaffer collaterals. To decipher the history of this well-known eponym, we review Schaffer's original German publication and follow the impact of his research in the contemporary literature.
Early stressful adverse situations may increase the vulnerability to cognitive deficits and psychiatric disorders, such as depression. Maternal separation (MS) has been used as an animal model to study changes in neurochemistry and behavior associated with exposure to early-life stress. This study investigated the effects of neonatal stress (MS) on the expression of synaptic plasticity markers in the hippocampus and a purported relationship to cognitive processes. Spatial learning (Morris water maze) significantly increased the expression of total levels of the neural cell adhesion molecule (NCAM), as well as its three major isoforms (NCAM-120, -140, and -180) both in the control and MS groups. Interestingly, these increases in NCAM expression after learning were lower in MS animals when compared with control rats. MS induced a significant decrease in total levels of NCAM, and specifically, in the NCAM-140 isoform expression. In the hippocampus of MS rats there was a significant decrease in brain-derived neurotrophic factor and synaptophysin mRNA densities. Cell proliferation, measured as BrdU-positive cells, was also decreased in the dentate gyrus of MS rats. Altogether these results suggest that MS can alter normal brain development, providing a potential mechanism by which early environmental stressors may influence vulnerability to show cognitive impairments later in life.
The cyclic AMP (cAMP)-response element binding protein (CREB) is an activity-dependent transcription factor that plays a role in synaptic plasticity and memory storage in Aplysia, Drosophila, and rodents. Mice with targeted deletions of two CREB isoforms (alpha and delta; CREB alphadelta mice) have been characterized on a mixed genetic background of C57BL/6 (B6) and 129/SvEv (129), as well as on a defined F1 hybrid of B6 and FVB/N, and these results suggest that the phenotype of CREB alphadelta mice depends critically on genetic background. In an examination of the hypothesis that the role of CREB in learning and memory can be influenced by strain differences, we analyzed mice with the CREB alphadelta mutation on an F1 hybrid background of B6 and 129 strains. CREB alphadelta mice on this background had impaired short-term and long-term cued and contextual fear conditioning and normal spatial learning in the Morris water maze. Our results suggest that at least some aspects of hippocampal function are normal in CREB alphadelta mice, and that CREB alphadelta mice on the B6/129 F1 background have alterations in amygdala function. These studies underscore the importance of controlling for genetic background in the behavioral analysis of knockout and transgenic mice.
Firing of place cells in the exploring rat conveys doubly coded spatial information: both the rate of spikes and their timing relative to the phase of the ongoing field theta oscillation are correlated with the location of the animal. Specifically, the firing rate of a place cell waxes and wanes, while the timing of spikes precesses monotonically as the animal traverses the portion of the environment preferred by the cell. We propose a mechanism for the generation of this firing pattern that can be applied for place cells in all three hippocampal subfields and that encodes spatial information in the output of the cell without relying on topographical connections or topographical input. A single pyramidal cell was modeled so that the cell received rhythmic inhibition in phase with theta field potential oscillation on the soma and was excited on the dendrite with input depending on the speed of the rat. The dendrite sustained an intrinsic membrane potential oscillation, frequency modulated by its input. Firing probability of the cell was determined jointly by somatic and dendritic oscillations. Results were obtained on different levels of abstraction: a purely analytical derivation was arrived at, corroborated by numerical simulations of rate neurons, and an extension of these simulations to spiking neurons was also performed. Realistic patterns of rate and temporal coding emerged and were found to be inseparable. These results may have implications on the robustness of information coding in place cell firing and on the ways information is processed in structures downstream to the hippocampus.
Phorbol esters, which activate protein kinase C (PKC), enhance synaptic transmission in the CA1 subfield of hippocampus, both in situ and in vitro. The increase in synaptic transmission could be the consequence of enhanced Ca influx into nerve terminals, and perhaps a more general increase in voltage-dependent Ca currents. The effects of phorbol 12,13-diacetate (PDAc) on the high-voltage activated (HVA) Ca currents, as well as spontaneous transient currents were therefore investigated by intracellular recording in hippocampal slices. PDAc selectively augmented, by 45% +/- 10%, the early peak of the HVA Ca current (but not its sustained component), and also spontaneous inhibitory postsynaptic currents. The inactive phorbol ester, 4 alpha-PDAc, had no comparable effects. The actions of PDAc were reversible on prolonged washing, and they were antagonized by the PKC inhibitors (1-(5-isoquinolinesulfonyl)-2-methyl piperazine (H-7) and monosialoganglioside (GM1). In addition, GM1, which also activates the Ca/calmodulin-dependent kinase, enhanced spontaneous excitatory postsynaptic currents, while inhibiting the IPSCs. It is concluded that activation of PKC increases HVA (probably N-type) Ca current and facilitates ongoing GABAergic IPSCs.
The effects of chronic exposure to cannabinoids on short-term memory in rats were assessed during repeated daily injections of an initially debilitating dose (3.75 mg/kg) of the potent CB1 cannabinoid receptor ligand, WIN 55,212-2. Delayed nonmatch to sample (DNMS) performance was assessed over a 35-day exposure period in which performance was initially disrupted during the first 21 days of exposure but recovered by day 30 and was stable at pre-drug levels for 5 days thereafter. Withdrawal was precipitated by injections of the CB1 receptor antagonist SR141716A and transiently reduced performance for 2 days but was restabilized to pre-drug levels within 3-4 days. Concomitant recording from identified CA1 and CA3 hippocampal neurons demonstrated a marked correspondence in the time course of suppression of peak firing in the sample and delay phases of the task to the drug-induced performance deficits over the same days of exposure. Hippocampal encoding of task-relevant events and performance levels "tracked" each other on a daily basis throughout the chronic cannabinoid treatment and withdrawal regimen. However, hippocampal neuronal activity in the nonmatch phase of the task was unaffected by the chronic cannabinoid treatment or withdrawal, suggesting that only a select population of hippocampal neurons and synapses are involved in cannabinoid-sensitive short-term memory processes.
Transcripts of primary and mature-miR-132 progressively increase during postnatal development in various brain regions. (A) Typical qPCR amplification plot of hippocampal mature-miR-132 levels expressed during postnatal Days 10, 17, and 24. (B–D) Relative levels of primary (left y-axis) and mature mir-132 (right y-axis) expression levels in the hippocampus, olfactory bulb, and striatum are depicted for the indicated developmental time points (n 5 5–6 mice/group, analyzed in triplicate). One-way ANOVA **P < 0.001 (Tukey's post hoc test).  
Activity-dependent changes in gene-expression are believed to underlie the molecular representation of memory. In this study, we report that in vivo activation of neurons rapidly induces the CREB-regulated microRNA miR-132. To determine if production of miR-132 is regulated by neuronal activity its expression in mouse brain was monitored by quantitative RT-PCR (RT-qPCR). Pilocarpine-induced seizures led to a robust, rapid, and transient increase in the primary transcript of miR-132 (pri-miR-132) followed by a subsequent rise in mature microRNA (miR-132). Activation of neurons in the hippocampus, olfactory bulb, and striatum by contextual fear conditioning, odor-exposure, and cocaine-injection, respectively, also increased pri-miR-132. Induction kinetics of pri-miR-132 were monitored and found to parallel those of immediate early genes, peaking at 45 min and returning to basal levels within 2 h of stimulation. Expression levels of primary and mature-miR-132 increased significantly between postnatal Days 10 and 24. We conclude that miR-132 is an activity-dependent microRNA in vivo, and may contribute to the long-lasting proteomic changes required for experience-dependent neuronal plasticity.
The central nervous system (CNS) exhibits remarkable plasticity in early life and can be altered significantly by various prenatal influences. We previously showed that prenatal exposure to morphine altered kinetic properties of N-methyl-D-aspartate (NMDA) receptor-mediated synaptic currents in the hippocampus of young rat offspring at the age of 14 days (P14). The present study further investigates whether NMDA receptor-mediated synaptic plasticity and/or cyclic adenosine monophosphate-responsive element-binding protein (CREBSerine-133), an important transcription factor underlying learning and memory, can be altered by prenatal morphine exposure in these offspring. Subsequently, the Morris water maze task was performed at the older ages (P28-P31). The magnitude of long-term depression (LTD) generated by a low-frequency stimulation (LFS, 1 Hz for 15 min) in hippocampal slices from the vehicle-control offspring (P14) was significantly larger than that in slices from the morphine-treated offspring, although there was no such difference in the magnitude of long-term potentiation (LTP) elicited by a high-frequency stimulation (100 Hz for 1 s) between the two groups. Comparison of the expression range of glutamatergic synaptic plasticity in slices from the vehicle-control and morphine-treated offspring, calculated as the difference in the maximal magnitude between LTP and LTD, demonstrated a remarkably smaller range in the slices from the morphine-treated offspring. In addition, the decreased phosphorylation of CREBSerine-133 and the impaired ability of spatial learning were also seen in the morphine-treated offspring, as compared with the vehicle-control offspring. Collectively, the study suggests that maternal exposure to morphine reduces the range of synaptic plasticity by decreasing the expression of LTD, but not of LTP, in CA1 pyramidal neurons of the hippocampus from rat offspring. More importantly, decreased phosphorylation of CREBSerine-133 may play a role for the impaired spatial learning and memory in rat offspring exposure to prenatal morphine. Thus, the findings here may provide important insights into cellular/molecular mechanisms underlying pathophysiological changes in the CNS of young offspring from morphine-addicted mothers and serve as a basis for possible therapeutic intervention.
Recent observations have caused a drastic shift in the conception of the hippocampus as a homogeneous structure that subserves cognitive functions, either spatial maps or short term episodic memory, to a structure that is associated with both cognitive and emotional functions. In fact, the assignment of cognitive functions to the hippocampus is restricted to its dorsal sector. In contrast, the ventral hippocampus (VH) appears to be associated with control of behavioral inhibition, stress and emotional memory, but not with strictly cognitive functions. Curiously, the VH but not the dorsal hippocampus (DH) is associated with the development of affective disorders. In line with these collective observations, we and others have found that the ability to evoke a sustained long term potentiation (LTP), a cellular correlate of learning and memory, is much lower in the VH compared to the DH. Strikingly, acute stress as well as direct exposure to corticosterone affect DH and VH in an opposite manner; causing facilitation of LTP in the VH and its suppression in the DH. This double dissociative action results from activation of different steroid receptor species in the DH and VH. Since the DH and VH differ in efferent connectivity, and since the strength of LTP can be considered as an indicator of strength of synaptic connectivity, these results suggest that stress regulates the routes by which the hippocampus is functionally linked to the rest of the brain such that under stress, the ventral route to the amygdala is enabled while the dorsal route to the neocortex is suppressed. This selective routing may underlie the complex outcome of stress on hippocampal and amygdala physiology and behavior.
The differentiation between the CA3 and CA1 fields of the mammalian hippocampus is one of the salient traits that set it apart from the organization of the homologue medial wall in reptiles and birds. CA3 is widely thought to function as an autoassociator, but what do we need CA1 for? Based on evidence for a specific role of CA1 in temporal processing, I have explored the hypothesis that the differentiation between CA3 and CA1 may help solve a computational conflict. The conflict is between pattern completion, or integrating current sensory information on the basis of memory, and prediction, or moving from one pattern to the next in a stored sequence. CA3 would take care of the former, while CA1 would concentrate on the latter. I have found the hypothesis to be only weakly supported by neural network simulations. The conflict indeed exists, but two mechanisms that would relate more directly to a functional CA3-CA1 differentiation were found unable to produce genuine prediction. Instead, a simple mechanism based on firing frequency adaptation in pyramidal cells was found to be sufficient for prediction, with the degree of adaptation as the crucial parameter balancing retrieval with prediction. The differentiation between the architectures of CA3 and CA1 has a minor but significant, and positive, effect on this balance. In particular, for a fixed anticipatory interval in the model, it increases significantly the information content of hippocampal outputs. There may therefore be just a simple quantitative advantage in differentiating the connectivity of the two fields. Moreover, different degrees of adaptation in CA3 and CA1 cells were not found to lead to better performance, further undermining the notion of a functional dissociation.
The hippocampus is an essential neural structure for spatial memory. Computational models suggest that the CA3 subregion of the hippocampus plays an essential role in encoding and retrieval of spatial memory. The perforant path (PPCA3) and dentate gyrus (DG)-mediated mossy fibers (MFs) compose major afferent inputs into CA3. A possible functional dissociation between these afferent inputs was attempted using a simple navigation test (i.e., the modified Hebb-Williams maze). Behavioral testing was combined with electrolytic lesions of PPCA3 or neurotoxic lesions of the DG, to eliminate each afferent input into CA3. Lesions in either afferent input into CA3 affected learning of an effective navigational path on the maze. The contributions of the two CA3 afferent inputs, however, were different regarding encoding and retrieval of memory measured based on indices operationally defined for the behavioral paradigm (i.e., encoding, the number of errors reduced within a day; retrieval, the number of errors reduced between days). The DG-lesioned animals exhibited deficits regarding the encoding index, but not the retrieval index, whereas the PPCA3-lesioned rats displayed deficits regarding the retrieval index, but not the encoding index. The results suggest that the two major afferent inputs of CA3 may contribute differentially to encoding and retrieval of spatial memory.
Growing evidence indicates that the amygdala modulates hippocampal functions. To test the hypothesis that this modulation may involve long-lasting effects on interneuronal networks in the hippocampus, changes in the expression of neurochemical markers specific for different interneuronal subpopulations were assessed in adult rats 96 h following acute infusion of low doses of the GABAA receptor antagonist picrotoxin into the amygdala. The numerical density (Nd) of somata showing immunoreactivity (IR) for parvalbumin (PVB) was decreased in dentate gyrus (DG) and the CA4-2 region, while that of calretinin (CR)-IR was decreased in DG and CA2. The Nd of calbindin D28k (CB)-IR somata was decreased in CA3-2. The densities of axon terminals arising from PVB-IR and cholecystokinin (CCK)-IR basket neurons were also altered, with those of CCK-IR terminals increased across all sectors, while PVB-IR terminals were decreased only in the CA region. Increases in CCK-IR terminals were paralleled by increases of terminals with IR for the 65-kD isoform of glutamate decarboxylase (GAD65). Mixed-effects statistical models, adapted specifically for these analyses, indicated that perturbations of amygdalar inputs to the hippocampus significantly alter the drive that hippocampal PVB-, CR-, and CB-IR neurons within the dentate gyrus/CA4 region exercise on CCK-IR terminals within the same region as well as in CA3-1. These results suggest that amygdalar modulation of specific neuronal subpopulations may induce lasting and far-reaching changes in the hippocampus during normal functioning, as well as in diseases involving a disruption of amygdalar activity. In particular, changes in specific interneuronal markers within selective hippocampal sectors detected in the present results are strikingly similar to those reported in this region in schizophrenia. These similarities suggest that, in this disease, a disruption of GABAergic transmission within the amygdala may play a significant role in the induction of abnormalities in the hippocampus.
The over-activation of glutamate receptors can lead to excitotoxic cell death and is believed to be involved in the progression of neurodegenerative events in the vulnerable hippocampus. Here, we used an in vitro slice model to study toxicity produced in the hippocampus by the mitochondrial toxin 3-nitropropionic acid (3-NP). The organotypic slice cultures exhibit native cellular organization as well as dense arborization of neuronal processes and synaptic contacts. The hippocampal slices were exposed to 3-NP for 2-20 days, causing calpain-mediated breakdown of the spectrin cytoskeleton, a loss of pre- and postsynaptic markers, and neuronal atrophy. The N-methyl-D-aspartate (NMDA) receptor antagonist memantine reduced both the cytoskeletal damage and synaptic decline in a dose-dependent manner. 3-NP-induced cytotoxicity, as determined by the release of lactate dehydrogenase, was also reduced by memantine with EC50 values from 1.7 to 2.3 microM. Propidium iodide fluorescence and phase contrast microscopy confirmed memantine neuroprotection against the chronic toxin exposure. In addition, the protected tissue exhibited normal neuronal morphology in the major hippocampal subfields. These results indicate that antagonists of NMDA-type glutamate receptors are protective during the toxic outcome associated with mitochondrial dysfunction. They also provide further evidence of memantine's therapeutic potential against neurodegenerative diseases.
It has been proposed that declarative memories can be dependent on both an episodic and a semantic memory system. While the semantic system deals with factual information devoid of reference to its acquisition, the episodic system, characterized by mental time travel, deals with the unique past experience in which an event took place. Episodic memory is characteristically hippocampus-dependent. Place cells are recorded from the hippocampus of rodents and their firing reflects many of the key characteristics of episodic memory. For example, they encode information about "what" happens "where," as well as temporal information. However, when these features are expressed during an animal's behavior, the neuronal activity could merely be categorizing the present situation and could therefore reflect semantic memory rather than episodic memory. We propose that mental time travel is the key feature of episodic memory and that it should take a form, in the awake animal, similar to the replay of behavioral patterns of activity that has been observed in hippocampus during sleep. Using tasks designed to evoke episodic memory, one should be able to see memory reactivation of behaviorally relevant sequences of activity in the awake animal while recording from hippocampus and other cortical structures.
Single neuron activity was recorded in the granular layer of the fascia dentata in freely moving rats, while the animals performed a spatial "working" memory task on an eight-arm maze. Using recording methods that facilitate detection of units with low discharge rates, it was found that the majority (88%) of cells in this layer have mean rates below 0.5 Hz, with a minimum of 0.01 Hz or less. The remaining recorded cells exhibited characteristics typical of the theta interneurons found throughout the hippocampus. Based on several criteria including relative proportion and the relation of their evoked discharges to the population spike elicited by perforant path stimulation, it was concluded that the low-rate cells correspond to granule cells. Granule cells exhibited clear spatially and directionally selective discharge that was at least as selective as that of a sample of CA3 pyramidal cells recorded under the same conditions. Granule cells had significantly smaller place fields than pyramidal cells, and tended to have more discontiguous subfields. There was no spatial correlation among simultaneously recorded adjacent granule cells. Granule cells also exhibited burst discharges reminiscent of complex spikes from pyramidal cells while the animals sat quietly; however, the spike duration of granule cells was significantly shorter than CA3 pyramidal cell spike durations. Under conditions of environmental stability, granule cell place fields were stable for at least several days. Following occasional maze rotations relative to the (somewhat impoverished) visual stimuli of the recording room, granule cell place fields were maintained relative to the distal spatial cues; however, frequent rotations of the maze sometimes resulted in a shift in the reference frame to the maze itself. These observations indicate that granule cells of the fascia dentata provide their CA3 targets with a high degree of spatial information, in the form of a sparsely coded, distributed representation.
The entorhinal cortex (EC) serves a pivotal role in corticohippocampal interactions, but a complete description of its extrinsic connections has not been presented. Here, we have summarized the cortical, subcortical, and hippocampal connections of the lateral entorhinal area (LEA) and the medial entorhinal area (MEA) in the rat. We found that the targets and relative strengths of the entorhinal connections are strikingly different for the LEA and MEA. For example, the LEA receives considerably heavier input from the piriform and insular cortices, whereas the MEA is more heavily targeted by the visual, posterior parietal, and retrosplenial cortices. Regarding subcortical connections, the LEA receives heavy input from the amygdala and olfactory structures, whereas the MEA is targeted by the dorsal thalamus, primarily the midline nuclei and also the dorsolateral and dorsoanterior thalamic nuclei. Differences in the LEA and MEA connections with hippocampal and parahippocampal structures are also described. In addition, because the EC is characterized by bands of intrinsic connectivity that span the LEA and MEA and project to different septotemporal levels of the dentate gyrus, special attention was paid to the efferents and afferents of those bands. Finally, we summarized the connections of the dorsocaudal MEA, the region in which the entorhinal "grid cells" were discovered. The subregional differences in entorhinal connectivity described here provide further evidence for functional diversity within the EC. It is hoped that these findings will inform future studies of the role of the EC in learning and memory.
Extinction of fear conditioning in animals is an excellent model for the study of fear inhibition in humans. Substantial evidence has shown that extinction is a new learning process that is highly context-dependent. Several recovery effects (renewal, spontaneous recovery, and reinstatement) after extinction suggest that the contextual modulation of extinction is a critical behavioral mechanism underlying fear extinction. In addition, recent studies demonstrate a critical role for hippocampus in the context control of extinction. A growing body of evidence suggests that the hippocampus not only plays a role in contextual encoding and retrieval of fear extinction memories, but also interacts with other brain structures to regulate context-specificity of fear extinction. In this article, the authors will first discuss the fundamental behavioral features of the context effects of extinction and its underlying behavioral mechanisms. In the second part, the review will focus on the brain mechanisms for the contextual control of extinction.
Chronic stress may have different effects on hippocampal CA3 and CA1 neuronal morphology and function depending upon hormonal status, but rarely are manipulations of stress and gonadal steroids combined. Experiment 1 investigated the effects of chronic restraint and 17beta-estradiol replacement on CA3 and CA1 dendritic morphology and spatial learning in ovariectomized (OVX) female Sprague-Dawley rats. OVX rats were implanted with 25% 17beta-estradiol, 100% cholesterol, or blank silastic capsules and then chronically restrained (6h/d/21d) or kept in home cages. 17beta-Estradiol or cholesterol prevented stress-induced CA3 dendritic retraction, increased CA1 apical spine density, and altered CA1 spine shape. The combination of chronic stress and 17beta-estradiol facilitated water maze acquisition compared to chronic stress + blank implants and nonstressed controls + 17beta-estradiol. To further investigate the interaction between 17beta-estradiol and stress on hippocampal morphology, experiment 2 was conducted on gonadally intact, cycling female rats that were chronically restrained (6h/d/21d), and then euthanized at proestrus (high ovarian hormones) or estrus (low ovarian hormones). Cycling female rats failed to show chronic stress-induced CA3 dendritic retraction at either estrous phase. Chronic stress enhanced the ratio of CA1 basal spine heads to headless spines as found in experiment 1. In addition, proestrous rats displayed increased CA1 spine density regardless of stress history. These results show that 17beta-estradiol or cholesterol protect against chronic stress-induced CA3 dendritic retraction in females. These stress- and 17beta-estradiol-induced morphological changes may provide insight into how dendritic complexity and spine properties contribute to spatial ability.
Beta-amyloid (Abeta) is a neuro-peptide implicated in the pathogenesis of Alzheimer's disease (AD). Abeta-peptide is known to disrupt cellular processes, including synaptic plasticity. To date, the precise mechanisms leading to the Abeta-mediated impairment of normal neurophysiological function still remains elusive. A rise in the pro-inflammatory cytokine interleukin-1-beta (IL-1beta) has been previously reported, following Abeta peptide insult. IL-1beta in turn, activates a cascade of pro-apoptotic markers, gradually leading to cell death. In this work, we have investigated the possible protective effects of interleukin-1 receptor antagonist (IL-1ra) on the effects of Abeta-peptide on long-term potentiation (LTP) in the CA1 region of the rat hippocampus in vivo. We observed a significant depression of LTP in the group of animals that received intracerebroventricular (icv) injection of Abeta-peptide (1-40) compared with control animals injected with vehicle. Administration of IL-1ra alone (icv) also resulted in a depression of LTP; however, there was no change in the baseline synaptic response. Combined injection of Abeta(1-40) + IL-1ra caused an attenuation of the effects observed with Abeta(1-40) alone for a period of up to 15 min following LTP induction; rescuing post-tetanicpotentiation (PTP). Gradually however, EPSP-values declined to produce a level of LTP similar to that observed following treatment with Abeta(1-40) alone. These results suggest that the acute Abeta-mediated impairment of PTP and LTP may be partial as a result of activation of an inflammatory response and the release of IL-1beta. The attenuation of plasticity by IL-1ra alone supports the theory that low levels of IL-1beta are required for normal synaptic plasticity. The limited rescue of the Abeta-mediated effects on LTP, in the presence of IL-1ra, may represent the short half life found with this receptor antagonist in vivo.
Progenitor cells that endure in different regions of the CNS after the initial neurogenesis can be expanded in culture and used as a source of donor tissue for grafting in neurodegenerative diseases. However, the proliferation and differentiation characteristics of residual neural progenitor cells from distinct regions of the CNS are mostly unknown. This study elucidated the characteristics of progenitor cells that endure in the CA3 region of the hippocampus after neurogenesis, by in vitro analyses of cells that are responsive to epidermal growth factor (EGF) or fibroblast growth factor-2 (FGF-2) in the embryonic day 19 (E19) rat hippocampus. Isolated cells from the E19 CA3 region formed neurospheres in the presence of either EGF or FGF-2, but the yield of neurospheres was greater with FGF-2 exposure, Differentiation cultures revealed a greater yield of neurons from FGF-2 neurospheres (60%) than from EGF neurospheres (35%). Exposure to brain-derived neurotrophic factor (BDNF) enhanced the yield of neurons from EGF neurospheres but had no consequence on FGF-2 neurospheres. A large number of neurons from EGF/FGF-2 neurospheres demonstrated clearly palpable morphological features of CA3 pyramidal neurons and lacked gamma-aminobutyric acid (GABA) expression. However, a fraction of neurons (17-20%) from EGF/FGF-2 neurospheres expressed GABA, and exposure to BDNF increased the number of GABAergic neurons (30%) from EGF neurospheres. Neurons from EGF/FGF-2 neurospheres also contained smaller populations of calbindin- and calretinin-positive interneuron-like cells. Thus, progenitor cells responsive to FGF-2 are prevalent in the CA3 region of the E19 rat hippocampus and give rise to a greater number of neurons than progenitor cells responsive to EGF. However, both FGF-2- and EGF-responsive progenitor cells from E19 CA3 region are capable of giving rise to CA3 field-specific phenotypic neurons. These results imply that progenitor cells that persist in the hippocampus after neurogenesis remain regionally restricted and hence retain their ability to give rise to region-specific phenotypic neurons even after isolation and expansion in vitro.
Hippocampal atrophy is a characteristic and early feature of Alzheimer's disease. Volumetry of the hippocampus using T1-weighted magnetic resonance imaging (MRI) has been used not only to assess hippocampal involvement in different neurodegenerative diseases as a potential diagnostic biomarker, but also to understand the natural history of diseases, and to track changes in volume over time. Assessing change in structure circumvents issues surrounding interindividual variability and allows assessment of disease progression. Disease-modifying effects of putative therapies are important to assess in clinical trials and are difficult using clinical scales. As a result, there is increasing use of serial MRI in trials to detect potential slowing of atrophy rates as an outcome measure. Automated and yet reliable methods of quantifying such change in the hippocampus would therefore be very valuable. Algorithms capable of measuring such changes automatically have been developed and may be applicable to predict decline to a diagnosis of dementia in the future. This article details the progress in using MRI to understand hippocampal changes in the degenerative dementias and also describes attempts to automate hippocampal segmentation in these diseases.
Hippocampal function varies in a subregion-specific fashion: spatial processing is thought to rely on the dorsal hippocampus, whereas anxiety-related behavior relies more on the ventral hippocampus. During development, neurogenesis in the dentate gyrus (DG) proceeds along ventral to dorsal as well as suprapyramidal to infrapyramidal gradients, but it is unclear whether regional differences in neurogenesis are maintained in adulthood. Moreover, it is unknown whether young neurons in the adult exhibit subregion-specific patterns of activation. We therefore examined the magnitude of neurogenesis and the activation of young and mature granule cells in DG subregions in adult rats that learned a spatial water maze task, swam with no platform, or were left untouched. We found that both adult neurogenesis and granule cell activation, as defined by c-fos expression in the granule cell population as a whole, were higher in the dorsal than the ventral DG. In contrast, c-fos expression in adult-born granule cells, identified by PSA-NCAM or location in the subgranular zone, occurred at a higher rate in the opposite subregion, the ventral DG. Interestingly, c-fos expression in the entire granule cell population was equivalent in water maze-trained rats and swim control rats, but was increased in the young granule cells only in the learning condition. These results provide new evidence that hippocampally-relevant experience activates young and mature neurons in different DG subregions and with different experiential specificity, and suggest that adult-born neurons may play a specific role in anxiety-related behavior or other nonspatial aspects of hippocampal function.
The hippocampus is involved in declarative memory and produces new neurons throughout adulthood. Numerous experiments have been aimed at testing the possibility that adult neurogenesis is required for learning and memory. However, progress has been encumbered by the fact that abating adult neurogenesis usually affects other biological processes, confounding the interpretation of such experiments. In an effort to circumvent this problem, we used a reverse approach to test the role of neurogenesis in hippocampus-dependent learning, exploiting the low levels of adult neurogenesis in the MRL/MpJ strain of mice compared with other mouse strains. We observed that adult MRL/MpJ mice produce 75% fewer new neurons in the dentate gyrus than age-matched C57BL/6 mice. Learning-induced synaptic remodeling, spatial learning, and visual recognition learning were reduced in MRL/MpJ mice compared with C57BL/6 mice. When MRL/MpJ mice were allowed unlimited access to running wheels, neurogenesis along with spatial learning and visual recognition learning were increased to levels comparable to those in running C57BL/6 mice. Together, these results suggest that adult neurogenesis is correlated with spatial learning and visual recognition learning, possibly by modulating morphological plasticity in the dentate gyrus.
Conditioned responses in Experiment 1. (A) Average freezing and standard error (s.e.) bars during re-exposure to the context (5 min). HPC: rats with total hippocampal lesions; Sham: control rats with sham lesions; D: Delay Group; T30: Trace Group. (B) Mean freezing and s.e. in the lesioned and sham rats during the 10 s presentation of the tone, average of three presentations. (C) Pretone, tone, and post-tone response pattern, in blocks of 10 s, average of three presentations of the tone. Alt CX: Mean freezing during the first 5 min of altered context. PRE-CS: Mean freezing in the 10 s preceding the three tone presentations, including carry over of freezing from previous presentations. CS: Mean freezing during the tone. POST 10 to POST 50: Freezing after the tone, in blocks of 10 s. Arrow: expected time of shock in the Trace group. 
Intrahippocampal injection sites. (A) Photomicrographs of a brain section at Bregma 22.8 stained with thionin showing the track of the guide cannula and the site of injection (arrow) in the dorsal hippocampus as indicated by the schematic representation on the right. (B) Location of the injection sites in the dorsal hippocampus. Gray dots: aCSF injection sites; Black dots: muscimol injection sites. (C) Photomicrographs of a brain section at Bregma 25.6 showing the track of the guide cannula and the site of injection (arrow) in the ventral hippocampus. (D) Location of the injection sites in the ventral hippocampus. Diagrams adapted from Paxinos and Watson (1998), with distance from Bregma in millimeters. 
Trace conditioning relies on the maintained representation of a stimulus across a trace interval, and may involve a persistent trace of the conditioned stimulus (CS) and/or a contribution of contextual conditioning. The role of hippocampal structures in these two types of conditioning was studied by means of pretraining lesions and reversible inactivation of the hippocampus in rats. Similar levels of conditioning to a tone CS and to the context were obtained with a trace interval of 30 s. Neurotoxic lesions of the whole hippocampus or reversible muscimol inactivation of the ventral hippocampus impaired both contextual and tone freezing in both trace- and delay-conditioned rats. Dorsal hippocampal injections impaired contextual freezing and trace conditioning, but not delay conditioning. No dissociation between trace and contextual conditioning was observed under any of these conditions. Altogether, these data indicate that the ventral and dorsal parts of the hippocampus compute different aspects of trace conditioning, with the ventral hippocampus being involved in fear and anxiety processes, and the dorsal hippocampus in the temporal and contextual aspects of event representation.
To lesion the cholinergic input to the hippocampus, rats received injections of 192 IgG-saporin into the medial septum/vertical limb of the diagonal band (MS/VDB). The lesions produced near-total loss of choline acetyltransferase (ChAT)-positive neurons in the MS/VDB. The loss was accompanied, however, by only partial decreases (to 40% of control levels) in acetylcholine (ACh) release in the hippocampus. Moreover, ACh release in the hippocampus increased when lesioned and control rats were tested on a spontaneous alternation task, indicating that there was significant residual cholinergic function in the hippocampus. The lesions were sufficient to impair spontaneous alternation scores. However, this impairment could be reversed by either systemic or intra-hippocampal injections of the indirect cholinergic agonist, physostigmine, providing additional evidence of residual and effective cholinergic functions in the hippocampus of lesioned rats. Moreover, systemic injections of physostigmine at doses that produced mild tremors in control rats led to more severe tremors in the lesioned rats, suggesting upregulation of cholinergic mechanisms after saporin lesions, likely in brain areas other than the hippocampus. Thus, these findings provide evidence for decreases in cholinergic input to the hippocampus accompanied by deficits on a spontaneous alternation tasks. The findings also provide evidence for considerable residual cholinergic input to the hippocampus after saporin lesions of the MS/VDB. Together, the results suggest that 192 IgG-saporin lesions of the MS/VDB, using methods often employed, do not fully remove septohippocampal cholinergic input to the hippocampus but are nonetheless sufficient to produce impairments on a task impaired by hippocampal lesions.
The extent to which small ensembles of neighboring hippocampal neurons alter their spatial firing patterns concurrently in response to stimulus manipulations was examined in young adult rats as well as in aged rats with and without memory impairment. Recordings from CA1 and CA3 cells were taken as rats performed a spatial radial-maze task that employed prominent distal visual stimuli attached to dark curtains surrounding the maze and local cues on each maze arm provided by inserts with distinctive visual, tactile, and olfactory stimuli. To test the influence of the different stimulus subsets, the distal and local cues were rotated 90° in opposite directions (a Double Rotation). In response to this manipulation, place fields could maintain a fixed position to room coordinates, rotate with either the local or the distal cues, disappear, or new fields could appear. On average 79% of the cells within an ensemble responded in the same way, but only 37% of all ensembles were fully concordant. Typically discordant ensembles had place fields that rotated with one set of cues, whereas the other fields disappeared or new fields appeared. Ensembles in which the place fields rotated in two opposite directions were less frequent in young rats than would be expected by the occurrence of the individual responses, indicating selective competition between directly conflicting representations and ultimate suppression of one. These findings indicate that hippocampal neurons independently encode distinct subsets of the cues in a complex environment, although processing within the hippocampal network may actively reduce the simultaneous representation of conflicting orientation information. This kind of population activity might reflect the higher-order organization of new memories within an established knowledge framework or schema.
In the study of temporal lobe epilepsy (TLE) the characterization of genes expressed in the hippocampus is of central importance for understanding their roles in epileptogenic mechanisms. Although several large-scale studies on TLE gene expression have been reported, precise assignment of individual genes associated with this syndrome is still debatable. Here we investigated differentially expressed genes by comparison of mRNAs from normal and epileptic rat hippocampus in the pilocarpine model of epilepsy. For this we used a powerful EST sequencing methodology, ORESTES (Open Reading frame Expressed Sequence Tags), which generates sequence datasets enriched for mRNAs open reading frames (ORFs) rather than simple 5' and 3' ends of mRNAs. Analysis of our sequences shows that ORESTES readily enables the identification of epilepsy associated ORFs. PFAM analysis of protein motifs present in our ORESTES epilepsy database revealed diverse important protein family domains, such as cytoskeletal, cell signaling and protein kinase domains, which could be involved in processes underlying epileptogenesis. More importantly, we show that the expression of homer 1a, known to be coupled to mGluR and NMDA synaptic transmission, is associated with pilocarpine induced status epilepticus (SE). The combined use of the pilocarpine model of epilepsy with the ORESTES technique can significantly contribute to the identification of specific genes and proteins related to TLE. This is the first study applying a large-scale method for rapid shotgun sequencing directed to ORFs in epilepsy research.
Cocaine induces an increase in hippocampal and nucleus accumbens (Nac) serotonin (5-HT) concentration parallel to locomotor activation. Both effects can be modulated by systemic 5-HT(1A)-receptor agonism/antagonism. Given the contribution of the hippocampus to spontaneous behavioral activity, these observations suggest a role for hippocampal 5-HT as well in the modulation of cocaine effects on behavior. To determine the role of hippocampal 5-HT(1A)-receptors in cocaine effects on behavior and hippocampal 5-HT release, we used in vivo microdialysis in freely moving rats. The 5-HT(1A)-receptor agonist, 8-OH-DPAT (0, 0.1, 1 and 10 microM), was applied locally into the hippocampus by reversed dialysis followed by a cocaine (10 mg/kg) or saline i.p. injection. The hippocampal 5-HT(1A)-receptor activation attenuated cocaine-induced hyperlocomotion and rearing behavior dose-dependently. Parallel to that, the cocaine-induced 5-HT increase was attenuated dose-dependently in the hippocampus but was left unaffected in the Nac. The intra-hippocampal application of 8-OH-DPAT affected neither behavioral activity nor 5-HT concentration in the hippocampus and in the Nac. In accord with these findings, hippocampal 5-HT(1A)-receptors may not be directly involved in the regulation of spontaneous behavior or basal 5-HT concentration in the hippocampus and Nac. However, the results indicate an inhibitory role of hippocampal 5-HT(1A)-receptors in cocaine-induced hyperactivity and in the 5-HT increase evoked by cocaine in the hippocampus but not in the Nac.
Although the role of 1alpha,25-dihydroxyvitamin D3 in calcium homeostasis of bone tissue is clear, evidence of the involvement of vitamin D3 in the central nervous system functions is increasing. In fact, vitamin D3 regulates vitamin D receptor and nerve growth factor expression, modulates brain development, and reverses experimental autoimmune encephalomyelitis. Only few studies, however, address vitamin D3 effect on embryonic hippocampal cell differentiation. In this investigation, the HN9.10e cell line was used as experimental model; these cells, that are a somatic fusion product of hippocampal cells from embryonic day-18 C57BL/6 mice and N18TG2 neuroblastoma cells, show morphological and cytoskeletal features similar to their neuronal precursors. By this model, we have studied the time course of vitamin D3 localization in the nucleus and its effect on proteins involved in proliferation and/or differentiation. We found that the translocation of vitamin D3 from cytoplasm to the nucleus is transient, as the maximal nuclear concentration is reached after 10 h of incubation with (3)H-vitamin D3 and decreases to control values by 12 h. The appearance of differentiation markers such as Bcl2, NGF, STAT3, and the decrease of proliferation markers such as cyclin-1 and PCNA are late events. Moreover, physiological concentrations of vitamin D3 delay cell proliferation and induce cell differentiation of embryonic cells characterized by modification of soma lengthening and formation of axons and dendrites.
Cannabinoids regulate numerous physiological and pathological events like inflammation or neurodegeneration via CB(1) and CB(2) receptors. The mechanisms behind cannabinoid effects show a high variability and may also involve transient receptor potential channels (TRP) and N-type voltage-gated Ca(2+) channels (Ca(v) 2.2). In the present study we investigated the neuroprotective effects of the synthetic cannabinoid WIN 55,212-2 (WIN) on dentate gyrus (DG) granule cells and elucidated the involvement of TRP and Ca(v) 2.2 that are shown to participate in inflammatory processes. Organotypic hippocampal slice cultures were excitotoxically lesioned using NMDA and subsequently incubated with different WIN concentrations (0.001-10 μM). WIN showed neuroprotective properties in an inverse concentration-dependent manner, most effectively at 0.01 μM. The CB(1) receptor antagonist AM251 blocked neuroprotection mediated by WIN whereas the CB(2) receptor antagonist AM630 showed no effects. Application of the TRPA1 blocker HC-030031 enhanced the neuroprotective efficacy of high (10 μM) WIN concentrations and the number of degenerating neurons became equal to that seen after application of the most effective WIN dose (0.01 μM). In contrast, the application of TRPA1 agonist icilin or allyl isothiocyanate (AITC) led to a stronger neurodegeneration. The use of TRPV1 blocker 6-iodo-nordihydrocapsaicin did not affect WIN-mediated neuroprotection. The selective Ca(v) 2.2 blocker ω-conotoxin (GVIA) completely blocked neuroprotection shown by 10 μM WIN. GVIA and HC-030031 exerted no effects at WIN concentrations lower than 10 μM. Our data show that WIN protects dentate gyrus granule cells in a concentration dependent manner by acting upon CB(1) receptors. At high (10 μM) concentrations WIN additionally activates TRPA1 and Ca(v) 2.2 within the hippocampal formation that both interfere with CB(1) receptor-mediated neuroprotection. This leads to the conclusion that physiological and pharmacological effects of cannabinoids strongly depend on their concentration and the neuroprotective efficacy of cannabinoids may be determined by interaction of activated CB(1) receptor, TRPA1, and Ca(v) 2.2.
Rats treated with low dose irradiation, to inhibit adult hippocampal neurogenesis, and control rats were administered a non-matching-to-sample (NMTS) task, which measured conditional rule learning and memory for specific events, and a test of fear conditioning in which a discrete CS was paired with an aversive US in a complex environment. Irradiated rats were impaired on the NMTS task when the intervals between sample and test trials were relatively long, and in associating the shock-induced fear with contextual cues in the fear conditioning task. Irradiated rats were not impaired in learning the basic NMTS rule or in performing that task when the intervals between the sample and test trials were short. Nor were there group differences in conditioning the fear response to the CS in the fear conditioning task. The results, which extend the range of hippocampus-dependent tasks that can be said to be vulnerable to the effects of neurogenesis suppression, support the hypothesis that new hippocampal cells generated in adulthood participate in a broad range of hippocampal functions.
In celebration of the 2014 Nobel Prize in Physiology or Medicine, this issue of Hippocampus includes a collection of commentaries from a broad range of perspectives on the significance of position coding neurons in the hippocampal region. From the perspective of this student of hippocampal physiology, it is argued that place cells and grid cells reflect the outcome of experiments that strongly select the information available and correspondingly observe singular "trigger features" of these neurons. Notably, however, in more naturalistic situations where multiple dimensions of information are available, hippocampal neurons have mixed selectivity wherein population-firing patterns reflect the organization of many features of experience. Thus, while discoveries on position coding were major breakthroughs in penetrating the hippocampal code, future studies exploring more complex behaviors hold the promise of revealing the full contribution of the hippocampal region to cognition and memory. This article is protected by copyright. All rights reserved. © 2015 Wiley Periodicals, Inc.
The award of the Nobel Prize in Medicine or Physiology in 2014 for the discovery of place and grid cells was both a personal award to three great scientists and also a mark of the maturity of systems neuroscience as a discipline. This article offers both personal and scientific reflections on these discoveries, detailing both how getting to know all three winners had an impact on my life and the research questions we shared in common work together. It ends with brief reflections on three important outstanding questions. This article is protected by copyright. All rights reserved. © 2015 Wiley Periodicals, Inc.
Episodic memory and semantic memory are two types of declarative memory. There have been two principal views about how this distinction might be reflected in the organization of memory functions in the brain. One view, that episodic memory and semantic memory are both dependent on the integrity of medial temporal lobe and midline diencephalic structures, predicts that amnesic patients with medial temporal lobe/diencephalic damage should be proportionately impaired in both episodic and semantic memory. An alternative view is that the capacity for semantic memory is spared, or partially spared, in amnesia relative to episodic memory ability. This article reviews two kinds of relevant data: 1) case studies where amnesia has occurred early in childhood, before much of an individual's semantic knowledge has been acquired, and 2) experimental studies with amnesic patients of fact and event learning, remembering and knowing, and remote memory. The data provide no compelling support for the view that episodic and semantic memory are affected differently in medial temporal lobe/diencephalic amnesia. However, episodic and semantic memory may be dissociable in those amnesic patients who additionally have severe frontal lobe damage.
Nonchimeric polytransgenic 152F7 mice encompassing four human chromosome 21 genes (DSCR3, DSCR5, TTC3, and DYRK1A) within the Down syndrome critical region present with learning and memory impairment. However, no abnormalities were shown by in vitro electrophysiological or neuroanatomical findings in hippocampus of 152F7 mice. To search for molecular changes that may be linked to cognitive impairment, we compared hippocampal protein levels between nontransgenic (WT) and 152F7 mice by a proteomic approach. Protein extracts were run on two-dimensional gel electrophoresis, protein spots were analyzed by mass spectrometry (MALDI-TOF-TOF) followed by quantification by specific software. Three hundred and nineteen different gene products were identified, and 48 proteins were assigned as signaling-related proteins. Stringent statistical analysis considering P < 0.005 as statistically significant based upon multiple testing revealed that growth factor receptor-bound protein 2 (Grb2) levels were decreased and an expression form of fascin 1 was increased in 152F7 mice when compared with WT. A series of proteins showed trends for increased and decreased hippocampal levels (P > 0.005 and P < 0.05). Only 2 out of 319 different gene products were dysregulated, pointing to the specificity of the analysis. Decreased Grb2 levels in the hippocampus of 152F7 mice may contribute to impaired cytoskeleton functions because dynamin 1 binds to Grb2 and involved in the formation of the endocytic process. Fascin dysregulation is of relevance for actin bundling in vesicle trafficking and may represent or lead to impaired neurotransmission that, in turn, may lead to the cognitive defect observed in this mouse model of Down syndrome.
The hippocampus is a key brain structure for the encoding of new experiences and environments. Hippocampal activity shows distinct oscillatory patterns, but the relationships between oscillations and memory are not well understood. Here we describe bursts of hippocampal approximately 23-30 Hz (beta2) oscillations in mice exploring novel, but not familiar, environments. In marked contrast to the relatively invariant approximately 8 Hz theta rhythm, beta2 power was weak during the very first lap of the novel environment, increased sharply as the mice reencountered their start point, then persisted for only a few minutes. Novelty-evoked oscillations reflected precise synchronization of individual neurons, and participating pyramidal cells showed a selective enhancement of spatial specificity. Through focal viral manipulations, we found that novelty-evoked oscillations required functional NMDA receptors in CA3, a subregion critical for fast oscillations in vitro. These findings suggest that beta2 oscillations indicate a hippocampal dynamic state that facilitates the formation of unique contextual representations.
Top-cited authors
Menno P Witter
  • Norwegian University of Science and Technology
Gyorgy Buzsáki
  • NYU Langone Medical Center
Raymond P Kesner
  • University of Utah
Hans Markowitsch
  • Bielefeld University
David Amaral
  • University of California, Davis