Spatial coding and physiological properties of hippocampal neurons in the Cornu Ammonis subregions

It is well‐established that the feed‐forward connected main hippocampal areas, CA3, CA2, and CA1 work cooperatively during spatial navigation and memory. These areas are similar in terms of the prevalent types of neurons; however, they display different spatial coding and oscillatory dynamics. Understanding the temporal dynamics of these operations requires simultaneous recordings from these regions. However, simultaneous recordings from multiple regions and subregions in behaving animals have become possible only recently. We performed large‐scale silicon probe recordings simultaneously spanning across all layers of CA1, CA2, and CA3 regions in rats during spatial navigation and sleep and compared their behavior‐dependent spiking, oscillatory dynamics and functional connectivity. The accuracy of place cell spatial coding increased progressively from distal to proximal CA1, suddenly dropped in CA2, and increased again from CA3a toward CA3c. These variations can be attributed in part to the different entorhinal inputs to each subregions, and the differences in theta modulation of CA1, CA2, and CA3 neurons. We also found that neurons in the subregions showed differences in theta modulation, phase precession, state‐dependent changes in firing rates and functional connectivity among neurons of these regions. Our results indicate that a combination of intrinsic properties together with distinct intra‐ and extra‐hippocampal inputs may account for the subregion‐specific modulation of spiking dynamics and spatial tuning of neurons during behavior. © 2016 Wiley Periodicals, Inc.     

Redefining the boundaries of the hippocampal CA2 subfield in the mouse using gene expression and 3-dimensional reconstruction

The morphology of neurons in the main divisions of the hippocampal complex allow the easy identification of granule cells in the dentate gyrus and pyramidal cells in the CA1 and CA3 regions of Ammon's horn. However, neurons in the CA2 subfield have been much more difficult to reliably identify. We have recently identified a set of genes whose expression is restricted to either the dentate gyrus, CA1, CA2, or CA3. Here we show that these genes have an essentially nonoverlapping distribution throughout the entire septotemporal extent of the hippocampus. 3-Dimensional reconstruction of serial sections processed for in situ hybridization of mannosidase 1, alpha (CA1), bcl-2-related ovarian killer protein (CA3), and Purkinje cell protein 4 (dentate gyrus+CA2) was used to define the boundaries of each subregion throughout the entire hippocampus. The boundaries observed for these three genes are recapitulated across a much larger set of genes similarly enriched in specific hippocampal subregions. The extent of CA2 defined on the basis of gene expression is somewhat larger than that previously described on the basis of structural anatomical criteria, particularly at the rostral pole of the hippocampus. These results indicate that, at least at the molecular level, there are robust, consistent genetic boundaries between hippocampal subregions CA1, CA2, CA3, and the dentate gyrus, allowing a redefinition of their boundaries in order to facilitate functional studies of different neuronal subtypes in the hippocampus.     

Test–retest reliability and longitudinal analysis of automated hippocampal subregion volumes in healthy ageing and Alzheimer's disease populations

The hippocampal formation is a complex brain structure that is important in cognitive processes such as memory, mood, reward processing and other executive functions. Histological and neuroimaging studies have implicated the hippocampal region in neuropsychiatric disorders as well as in neurodegenerative diseases. This highly plastic limbic region is made up of several subregions that are believed to have different functional roles. Therefore, there is a growing interest in imaging the subregions of the hippocampal formation rather than modelling the hippocampus as a homogenous structure, driving the development of new automated analysis tools. Consequently, there is a pressing need to understand the stability of the measures derived from these new techniques. In this study, an automated hippocampal subregion segmentation pipeline, released as a developmental version of Freesurfer (v6.0), was applied to T1‐weighted magnetic resonance imaging (MRI) scans of 22 healthy older participants, scanned on 3 separate occasions and a separate longitudinal dataset of 40 Alzheimer's disease (AD) patients. Test–retest reliability of hippocampal subregion volumes was assessed using the intra‐class correlation coefficient (ICC), percentage volume difference and percentage volume overlap (Dice). Sensitivity of the regional estimates to longitudinal change was estimated using linear mixed effects (LME) modelling. The results show that out of the 24 hippocampal subregions, 20 had ICC scores of 0.9 or higher in both samples; these regions include the molecular layer, granule cell layer of the dentate gyrus, CA1, CA3 and the subiculum (ICC > 0.9), whilst the hippocampal fissure and fimbria had lower ICC scores (0.73–0.88). Furthermore, LME analysis of the independent AD dataset demonstrated sensitivity to group and individual differences in the rate of volume change over time in several hippocampal subregions (CA1, molecular layer, CA3, hippocampal tail, fissure and presubiculum). These results indicate that this automated segmentation method provides a robust method with which to measure hippocampal subregions, and may be useful in tracking disease progression and measuring the effects of pharmacological intervention.     

Effects of discrete kainic acid-induced hippocampal lesions on spatial and contextual learning and memory in rats

Substantial information is available concerning the influence of global hippocampal lesions on spatial learning and memory, however the contributions of discrete subregions within the hippocampus to these functions is less well understood. The present investigation utilized kainic acid to bilaterally lesion specific areas of the rat hippocampus. These animals were subsequently tested on a spatial orientation task using a circular water maze, and on an associative/contextual task using passive avoidance conditioning. The results indicate that both the dorsal CA1 and the ventral CA3 subregions play important roles in learning. Specifically, CA1 lesions produced a deficit in the acquisition of the water maze task and a significant memory impairment on the passive avoidance task. CA3 lesions also caused learning deficits in the acquisition of the water maze task, and produced even greater impairments in performance on the passive avoidance task. We conclude that CA1 and CA3 hippocampal subregions each play significant roles in the overall integration of information concerning spatial and associative learning.     

Parallel activation of field CA2 and dentate gyrus by synaptically elicited perforant path volleys

Previous studies showed that dorsal psalterium (PSD) volleys to the entorhinal cortex (ENT) activated in layer II perforant path neurons projecting to the dentate gyrus. The discharge of layer II neurons was followed by the sequential activation of the dentate gyrus (DG), field CA3, field CA1. The aim of the present study was to ascertain whether in this experimental model field, CA2, a largely ignored sector, is activated either directly by perforant path volleys and/or indirectly by recurrent hippocampal projections. Field potentials evoked by single-shock PSD stimulation were recorded in anesthetized guinea pigs from ENT, DG, fields CA2, CA1, and CA3. Current source-density (CSD) analysis was used to localize the input/s to field CA2. The results showed the presence in field CA2 of an early population spike superimposed on a slow wave (early response) and of a late and smaller population spike, superimposed on a slow wave (late response). CSD analysis during the early CA2 response showed a current sink in stratum lacunosum-moleculare, followed by a sink moving from stratum radiatum to stratum pyramidale, suggesting that this response represented the activation and discharge of CA2 pyramidal neurons, mediated by perforant path fibers to this field. CSD analysis during the late response showed a current sink in middle stratum radiatum of CA2 followed by a sink moving from inner stratum radiatum to stratum pyramidale, suggesting that this response was mediated by Schaffer collaterals from field CA3. No early population spike was evoked in CA3. However, an early current sink of small magnitude was evoked in stratum lacunosum-moleculare of CA3, suggesting the presence of synaptic currents mediated by perforant path fibers to this field. The results provide novel information about the perforant path system, by showing that dorsal psalterium volleys to the entorhinal cortex activate perforant path neurons that evoke the parallel discharge of granule cells and CA2 pyramidal neurons and depolarization, but no discharge of CA3 pyramidal neurons. Consequently, field CA2 may mediate the direct transfer of ENT signals to hippocampal and extrahippocampal structures in parallel with the DG-CA3-CA1 system and may provide a security factor in situations in which the latter is disrupted.     

Hippocampal neuron and glial cell numbers in Parkinson's disease--a stereological study

Hippocampal atrophy and neuron loss are early and reproducible findings in Alzheimer's disease, and recent magnetic resonance imaging studies indicate that hippocampal atrophy may also be present in Parkinson's disease (PD). To determine whether or not cell loss occurs in PD, we estimated the total neuron and glial cell numbers as well as the total volume unilaterally in the hippocampi of eight demented PD patients and eight control subjects. Cell numbers were estimated in the neuron-containing layers of CA1, CA2-(3), CA4, the dentate gyrus, and subiculum using the optical-fractionator technique. The Cavalieri method was used to estimate the volume of the total hippocampus and its subregions. We did not find significant differences in cell numbers or volumes in PD brains when compared with control subjects. Our results thus indicate that hippocampal atrophy and cell loss are not necessarily involved in the memory impairment and dementia observed in PD.     

Spatio-temporal Distribution of Epileptiform Potentials in the Hippocampal Slice: Recordings with Voltage-sensitive Dyes

Voltage-sensitive dyes and fast optical recording techniques were used to monitor the spatio-temporal activity pattern of epileptiform potentials in hippocampal slices from guinea pigs. Epileptiform potentials were induced by adding 4-aminopyridine to the bath solution and applying single pulse stimulation either to the stratum pyramidale of area CA3 or to the stratum radiatum of area CA1. Optical activity as well as intra- or extracellular electrical activity were recorded from area CA1. There was a good correlation between optical and intracellular records from the same site. The spatio-temporal activity pattern of control and epileptiform potentials elicited by stimulation of CA1 was similar for the initial part of the potential. Then, epileptiform changes became apparent throughout the vertical extent of pyramidal neurons. Qualitative changes occurred in the stratum moleculare, reflecting activity of apical dendrites, such changes occurred even more strongly in the stratum oriens, reflecting activity of basal dendrites. The activity in the stratum oriens occurred relatively late, so that it cannot account for the initiation of epileptic discharges. It might, however, play an important role in the synchronization and spread of epileptiform potentials. The investigation of the horizontal distribution of potentials throughout the entire area CA1 indicates that different mechanisms are involved in the spread of epileptiform activity elicited by stimulation of CA1 and stimulation of CA3.     

Differential contributions of dorsal hippocampal subregions to memory acquisition and retrieval in contextual fear-conditioning

The hippocampus is an essential neural structure in developing contextual memory in a situation in which rapid development of associative learning should occur. We tested a subregion-specific contribution in the hippocampus to memory acquisition and retrieval, using the contextual fear-conditioning paradigm. The current results suggest that all three subregions (i.e., CA3, CA1, and dentate gyrus) of the hippocampus contribute to rapid acquisition of contextual memory in the initial phase of acquisition. The involvement of CA3 seems to be important at the earliest stage of acquisition, presumably for developing instant representation of a context. The role of CA3, however, was minimal in retrieving contextual memory after a long time period (i.e., 24 h), whereas the other subregions (i.e., CA1 and dentate gyrus) were critically involved. The results indicate time-dependent differential contributions of the hippocampal subregions to memory acquisition and retrieval in contextual fear-conditioning.     

ACh对正常大鼠和吗啡成瘾大鼠海马CA1区痛反应电活动的影响

目的研究ACh对正常大鼠和吗啡成瘾大鼠海马CA1区痛兴奋神经元(pain-excitation neurons,PEN)和痛抑制神经元(pain-inhibitationneurons,PIN)电活动的影响,进一步探讨ACh对正常和吗啡成瘾状态下CA1区痛觉调制的作用及机制。方法电刺激坐骨神经作为伤害性电刺激,在细胞外用玻璃微电极记录CA1区PEN和PIN的放电,观察ACh对正常大鼠和吗啡成瘾大鼠CA1区PEN和PIN电活动的影响。结果伤害性刺激能够增强PEN的电活动,而减弱PIN的电活动。正常大鼠中,ACh使PEN的痛诱发放电频率降低,PIN的放电频率增加;ACh的作用在注射后4 min达到峰值。吗啡成瘾大鼠中,ACh同样也抑制了PEN的电活动,兴奋PIN的电活动,但是作用的高峰出现在注射后6min。胆碱能受体拮抗剂阿托品可阻断ACh的作用。结论海马CA1区内的胆碱能神经元和毒蕈碱受体参与了伤害性信息的处理,并且起到了镇痛作用。吗啡成瘾可以降低CA1区痛反应神经元对伤害性刺激的敏感性。     

Episodic autobiographical memory is associated with variation in the size of hippocampal subregions

Striking individual differences exist in the human capacity to recollect past events, yet, little is known about the neural correlates of such individual differences. Studies investigating hippocampal volume in relation to individual differences in laboratory measures of episodic memory in young adults suggest that whole hippocampal volume is unrelated (or even negatively associated) with episodic memory. However, anatomical and functional specialization across hippocampal subregions suggests that individual differences in episodic memory may be linked to particular hippocampal subregions, as opposed to whole hippocampal volume. Given that the DG/CA_2/3 circuitry is thought to be especially critical for supporting episodic memory in humans, we predicted that the volume of this region would be associated with individual variability in episodic memory. This prediction was supported using high‐resolution MRI of the hippocampal subfields and measures of real‐world (autobiographical) episodic memory. In addition to the association with DG/CA_2/3, we further observed a relationship between episodic autobiographical memory and subiculum volume, whereas no association was observed with CA₁ or with whole hippocampal volume. These findings provide insight into the possible neural substrates that mediate individual differences in real‐world episodic remembering in humans.     

Electrophysiological evidence for a rapid membrane action of the gonadal steroid, 17β-estradiol, on CA1 pyramidal neurons of the rat hippocampus

The rapid electrophysiological effects of 17β-estradiol on CA1 pyramidal neurons(n = 86) were investigated utilizing intracellular recording from the rat hippocampal slice preparation. Bath application of 17β-estradiol, but not 17α-estradiol, caused a reversible depolarization and increased input resistance with a latency of less than 1 min in 19.8% of CA1 neurons tested. There was no significant difference in the percentage of estradiol-responsive cells between male and female rats. Estradiol-responsive cells were identified from prepubertal female rats, as well as females in all stages of the estrous cycle. 17β-estradiol had no effect on the slow afterhyperpolarization or accommodative properties of CA1 neurons. In 2 out of 4 cells tested, the specific antiestrogen, tamoxifen, blocked the excitatory response to 17β-estradiol.     

Long-lasting effects of partial hippocampal kindling on hippocampal physiology and function

The objective of this project was to study the behavioral and physiological effects at 6–9 weeks after evoking 15 afterdischarges (ADs) in hippocampal CA1 (partial hippocampal kindling). Rats were trained on the open radial arm maze (RAM) with all eight arms baited, kindled, and then tested again on the RAM, followed by in vitro recordings at 8–9 weeks after kindling. Partial kindling was manifested by an increase in hippocampal AD duration. Enhancement of the commissural basal dendritic excitatory postsynaptic potential (EPSP) was observed for at least 1 day after the ADs kindled rats performed worse than control rats during the 1st but not during the 7th or 8th week after kindling. Rats that were slow in acquiring the RAM showed more RAM errors after kindling than those that showed fast acquisition. At 8–9 weeks after kindling, as shown by field potential recording in the hippocampal slice in vitro, kindled rats showed an increase in paired-pulse facilitation (PPF) of the EPSP in CA1 but a decreased PPF of the perforant path to dentate gyrus EPSP; no change in the PPF of the population spike was found in CA1 or DG. In a second group of rats that were not run on the RAM, at 6 weeks after kindling, PPF of the population EPSP and population spike were enhanced in the kindled rats compared to the control rats in CA1, but not in DG or CA3 in vitro (at 1.5, 2, or 4 times threshold intensity). In conclusion, partial hippocampal kindling induced persistent physiological effects for up to 8–9 weeks, and it is suggested that the normalization of the paired-pulse population spike response in CA1 and DG at more than 6 weeks after kindling may be accompanied by a recovery of RAM performance. © 1994 Wiley-Liss, Inc.     

Heterogeneity among hippocampal pyramidal neurons revealed by their relation to theta-band oscillation and synchrony

Intracellular recordings were made in the dorsal hippocampal formation of urethane-anesthetized rats as the local field activity spontaneously cycled between a synchronous condition termed theta and an asynchronous condition termed LIA. All cells reported in this study were labeled with Neurobiotin and classified as theta-related or non-theta-related according to the system of Colom and Bland [Colom, L.V., Bland, B.H., 1987. State-dependent spike train dynamics of hippocampal formation neurons: evidence for theta-ON and theta-OFF cells. Brain Res. 422; 277-286]. The findings are the first demonstration that hippocampal pyramidal cells are functionally heterogeneous in relation to the generation of theta-band oscillation and synchrony. In field CA1 pyramidal cells formed theta-related subsets of phasic theta-ON cells and tonic theta-ON cells and non-theta-related subsets of simple spike discharging cells, complex spike discharging cells and "silent" cells. Similar findings were evident for CA3 pyramidal cells.     

ACh对正常大鼠和吗啡成瘾大鼠海马CA1区痛反应电活动的影响

目的 研究ACh对正常大鼠和吗啡成瘾大鼠海马CA1区痛兴奋神经元(pain-excitation neurons,PEN)和痛抑制神经元(pain-inhibitation neurons,PIN)电活动的影响,进一步探讨ACh对正常和吗啡成瘾状态下CA1区痛觉调制的作用及机制.方法 电刺激坐骨神经作为伤害性电刺激,在细胞外用玻璃微电极记录CA1区PEN和PIN的放电,观察ACh对正常大鼠和吗啡成瘾大鼠CA1区PEN和PIN电活动的影响.结果 伤害性刺激能够增强PEN的电活动,而减弱PIN的电活动.正常大鼠中,ACh使PEN的痛诱发放电频率降低,PIN的放电频率增加;ACh的作用在注射后4 min达到峰值.吗啡成瘾大鼠中,ACh同样也抑制了PEN的电活动,兴奋PIN的电活动,但是作用的高峰出现在注射后6min.胆碱能受体拮抗剂阿托品可阻断ACh的作用.结论 海马CA1区内的胆碱能神经元和毒蕈碱受体参与了伤害性信息的处理,并且起到了镇痛作用.吗啡成瘾可以降低CA1区痛反应神经元对伤害性刺激的敏感性.     

Somatostatin hyperpolarizes hippocampal pyramidal cells in vitro

Superfusion onto hippocampal slices of low concentrations of the tetradecapeptide somatostatin (SS), or an SS analogue having CNS activity, reversibly hyperpolarized pyramidal neurons, as revealed by intracellular recording. The hyperpolarizations were accompanied by reductions in spontaneous and evoked spike discharge and in input resistance; the magnitude of the hyperpolarizations was not influenced by 10 mM MgCl₂ added to the perfusate to block synaptic transmission.     

Differential effects of kainic-acid-induced hippocampal lesions on the acquisition of the jump-box avoidance task in mice

This research examined the effects of kainic-acid (KA)-induced hippocampal lesions on the acquisition of a jump-box avoidance task in mice. When the hippocampal lesion was unilateral, mice with lesions in the lateral portion of the CA1 region showed deficits in acquisition while those with lesions in the CA3 region and/or the medial portion of the CA1 region did not. When the hippocampal lesion was bilateral, on the other hand, mice with lesions in the CA3 region and/or the lateral portion of the CA1 region were deficient in acquisition while those with lesions in the medial portion of the CA1 region were not. Among the mice with various KA-induced hippocampal lesions avoidance responses were not correlated with the level of open field activity, indicating that acquisition differences observed are not dependent upon, nor secondary to, levels of spontaneous activity. These results strongly suggest that there exist regional differences in the involvement of hippocampal pyramidal cell layers in acquisition of the jump-box avoidance task: both the CA3 region and the lateral portion of the CA1 region are involved in acquisition, while the medial portion of the CA1 region is not.     

Histamine modulates reactivity of hippocampal CA3 neurons to afferent stimulation in vitro

Intracellular activity was recorded from hippocampal CA3 pyramidal cells maintained in vitro. Histamine (HA) produced a slow depolarization associated with minimal conductance changes. In addition, there was an increase in action potential discharge rates and the emergence of bursting firing patterns. EPSP size increased by about 50% and spontaneous dendritic spikes were observed. These effects were markedly reduced by retrodotoxin. Extracellular recording of population spikes revealed a marked difference between CA1 and CA3 regions; in the former HA produced an increase in population spike size whereas in the latter this increase was larger and was associated with the appearance of secondary and tertiary population spikes. It is suggested that HA produces its effects by enhancing release of neurotransmitters from excitatory synapses on the recorded neurons.     

The role of the dentate gyrus, CA3a,b, and CA3c for detecting spatial and environmental novelty

It has been suggested that the dentate gyrus (DG) and CA3 cooperate to efficiently process spatial information. The DG has been proposed to be important for fine spatial discrimination, and the CA3 has been proposed to mediate larger scale spatial information processing. To evaluate the roles of the DG and CA3a,b for spatial processing, we developed a task that measures responses to either overall environmental novelty or a response to more subtle changes within the environment. Animals with lesions to the DG showed impaired novelty detection for both environment as well as smaller changes in the environment, whereas animals with lesions to CA3a,b showed no such deficits. A closer look at the lesions suggested that the CA3 lesions included only CA3a and CA3b, but spared CA3c. To test the role of the spared CA3c region, animals with selective lesions to CA3c that spared CA3a,b were run on the same task and showed an intermediate pattern of deficits. These results suggest that the DG is critical for spatial information processing. These data also suggest that CA3 is a heterogeneous structure, with CA3c lesioned animals showing greater spatial processing deficits than CA3a,b lesioned animals. These findings extend our knowledge of hippocampal function and need to be accounted for in future computational models.     

An analysis of cholinoceptive neurons in the hippocampal formation by direct microinfusion

Microinfusions of cholinergic agents were made in various sites in the dorsal hippocampal formation of urethane anaesthetized rats. Infusions of eserine or carbachol elicited hippocampal theta activity when made in areas containing high levels of cholinergic markers: the stratum oriens and radiatum of the CA1 and CA3, the stratum moleculare and stratum granulosum of the dentate gyrus and the infragranular region of the hilus. Subsequent infusions of atropine sulfate antagonized the theta activity. Control infusions of equal volumes of saline in active sites were without effect. Infusions of eserine or carbachol in the vicinity of the hippocampal fissure, the stratum lacunosum/moleculare of the CA1 or CA3, in the deep regions of the hilus, and in the lateral ventricle and overlying neocortex, were also without effect. Furthermore, in active sites, the latency to onset of theta and subsequent theta frequency, were both directly related to the total amount of carbachol infused. Thus, areas in which theta could be elicited with a cholinergic agonist (carbachol), or an anticholinesterase (eserine) and antagonized with atropine, were found to correspond well to areas previously found to contain a high density of cholinoceptive neurons, using autoradiographic and immunohistochemical techniques. These results provide further support for the involvement of acetylcholine as a neurotransmitter in the generation of type 2 theta in the hippocampal formation.