A role for hilar cells in pattern separation in the dentate gyrus: a computational approach

We present a simple computational model of the dentate gyrus to evaluate the hypothesis that pattern separation, defined as the ability to transform a set of similar input patterns into a less-similar set of output patterns, is dynamically regulated by hilar neurons. Prior models of the dentate gyrus have generally fallen into two categories: simplified models that have focused on a single granule cell layer and its ability to perform pattern separation, and large-scale and biophysically realistic models of dentate gyrus, which include hilar cells, but which have not specifically addressed pattern separation. The present model begins to bridge this gap. The model includes two of the major subtypes of hilar cells: excitatory hilar mossy cells and inhibitory hilar interneurons that receive input from and project to the perforant path terminal zone (HIPP cells). In the model, mossy cells and HIPP cells provide a mechanism for dynamic regulation of pattern separation, allowing the system to upregulate and downregulate pattern separation in response to environmental and task demands. Specifically, pattern separation in the model can be strongly decreased by decreasing mossy cell function and/or by increasing HIPP cell function; pattern separation can be increased by the opposite manipulations. We propose that hilar cells may similarly mediate dynamic regulation of pattern separation in the dentate gyrus in vivo, not only because of their connectivity within the dentate gyrus, but also because of their modulation by brainstem inputs and by the axons that "backproject" from area CA3 pyramidal cells.     

On the number of neurons in the dentate gyrus of the rat

We have estimated the number of dentate granule cells in Sprague-Dawley and Wistar rats at 1, 4 and 12 months of age. In Sprague-Dawley rats the number of granule cells is relatively constant throughout this period at about 1 million. In Wistar rats, on the other hand, there is a progressive increase in the number from about 700,000 at 1 month to 1 million at 4 months; thereafter the number declines to about 800,000 at 1 year. Estimates of the numbers of cells in the polymorphic zone that can be stained immunohistochemically for somatostatin, cholecystokinin, vasoactive-intestinal peptide, and glutamic acid decarboxylase show no appreciable differences in the two strains.     

Cholinergic innervation of the rat dentate gyrus: an immunocytochemical and electron microscopical study

Immunocytochemical studies using a monoclonal antibody to choline acetyltransferase (ChAT) were performed on sections of rat dentate gyrus. Light microscopical analysis of the immunoreactivity revealed dense fiber networks and many punctate structures predominantly located at the interface of the granule cell layer and molecular layer. In the elctron microscope, the immunostained punctate structures were identified as synaptic boutons which formed mainly symmetrical contacts onto dendritic elements. Few ChAT-immunoreactive boutons formed axosomatic contacts.     

Influence of neurons of the parafascicular region on neuronal transmission from perforant pathway through dentate gyrus

We have previously reported that activation of an ascending brainstem pathway by stimulation of the median raphe nucleus (MR) influences neuronal transmission from the perforant pathway through the dentate gyrus in a behaviorally dependent manner. In particular, stimulation of the MR markedly facilitated such transmission when applied during slow-wave sleep (SWS), but was ineffective when applied during the still-alert state (SAL). We present here evidence for a relay in this circuit located rostral to the MR in cells proximal to the fasciculus retroflexus (PF, parafascicular region). In contrast to stimulation of the MR, stimulation of the PF facilitates neuronal transmission from the perforant pathway through the dentate gyrus during both SWS and SAL indicating the presence of a gate at or proximal to the PF that is preferentially closed during SAL.     

Granule cells of the dentate gyrus with basal and recurrent dendrites in schizophrenic patients and controls. A comparative Golgi study

We report on structural variability of granule cells in the human dentate gyrus. Granule cells with basal and recurrent dendrites are a normal finding in the human brain. We detect 28.3% granule cells with basal dendrites in non-psychiatrically ill humans compared to rats (2%) and primates (10%). This can be seen as an indication for the higher phylogeny of the human brain. In addition we find a significantly higher incidence of granule cells with basal dendrites (45.7%) in brains of schizophrenic patients. Whereas drug influences during lifetime cannot fully be excluded, we tend to interpret this finding as a plastic reaction to prenatal developmental malformations of the impinging rostral entorhinal region.     

Guided saccades modulate object and face-specific activity in the fusiform gyrus

We investigated the influence of saccadic eye movements on the magnitude of functional MRI (fMRI) activation in brain regions known to participate in object and face perception. In separate runs, subjects viewed a static image of a uniform gray field, a face, or a flower. Every 500 ms a small fixation cross made a discrete jump within the image and subjects were required to make a saccade and fixate the cross at its new location. Each run consisted of alternating blocks in which the subject was guided to make small and large saccades. A comparison of large vs. small saccade blocks revealed robust activity in the oculomotor system, particularly within the frontal eye fields (FEF), intraparietal sulcus (IPS), and superior colliculi regardless of the background image. Activity within portions of the ventral occipitotemporal cortex (VOTC) including the lingual and fusiform gyri was also modulated by saccades, but here saccade-related activity was strongly influenced by the background image. Activity within the VOTC was strongest when large saccadic eye movements were made over an image of a face or a flower compared to a uniform gray image. Of most interest was activity in the functionally predefined face-specific region of the fusiform gyrus, where large saccades made over a face increased activity, but where similar large saccades made over a flower or a uniform gray field did not increase activity. These results demonstrate the potentially confounding influence of uncontrolled eye movements for neuroimaging studies of face and object perception.     

Axon arbors and synaptic connections of hippocampal mossy cells in the rat in vivo

The axon collateralization patterns and synaptic connections of intracellularly labeled and electrophysiologically identified mossy cells were studied in rat hippocampus. Light microscopic analysis of 11 biocytin-filled cells showed that mossy cell axon arbors extended through an average of 57% of the total septotemporal length of the hippocampus (summated two-dimensional length, not adjusted for tissue shrinkage). Axon collaterals were densest in distant lamellae rather than in lamellae near the soma. Most of the axon was concentrated in the inner one-third of the molecular layer, with the hilus containing an average of only 26% of total axon length and the granule cell layer containing an average of only 7%. Ultrastructural analysis was carried out on three additional intracellularly stained mossy cells, in which axon collaterals and synaptic targets were examined in serial sections of chosen axon segments. In the central and subgranular regions of the hilus, mossy cell axons established a low density of synaptic contacts onto dendritic shafts, neuronal somata, and occasional dendritic spines. Most hilar synapses were made relatively close to the mossy cell somata. At greater distances from the labeled mossy cell (1–2 mm along the septotemporal axis), the axon collaterals ramified predominantly within the inner molecular layer and made a high density of asymmetric synaptic contacts almost exclusively onto dendritic spines. Quantitative measurements indicated that more than 90% of mossy cell synaptic contacts in the ipsilateral hippocampus are onto spines of proximal dendrites of presumed granule cells. These results are consistent with a primary mossy cell role in an excitatory associational network with granule cells of the dentate gyrus. © 1996 Wiley-Liss, Inc.     

The histochemical localization of cytochrome oxidase in the dentate gyrus of the rat hippocampus

In the dentate gyrus of the rat's hippocampal formation, the activity of an oxidative enzyme, cytochrome oxidase, has been localized mostly to the molecular layer with histochemical methods that utilize diaminobenzidine. The electron microscopic localization of cytochrome oxidase indicated that mitochondria within granule cell dendrites were very reactive while those within the somata and mossy fiber terminals of this neuronal type were less reactive. Caution must be used when predicting the relative physiological activities of neurons with this method because differential activities of this enzyme occur within separate parts of the same neuronal population.     

Cannabinoid regulation of neurons in the dentate gyrus during epileptogenesis: Role of CB1R-associated proteins and downstream pathways

The hippocampal formation plays a central role in the development of temporal lobe epilepsy (TLE), a disease characterized by recurrent, unprovoked epileptic discharges. TLE is a neurologic disorder characterized by acute long-lasting seizures (i.e., abnormal electrical activity in the brain) or seizures that occur in close proximity without recovery, typically after a brain injury or status epilepticus. After status epilepticus, epileptogenic hyperexcitability develops gradually over the following months to years, resulting in the emergence of chronic, recurrent seizures. Acting as a filter or gate, the hippocampal dentate gyrus (DG) normally prevents excessive excitation from propagating through the hippocampus, and is considered a critical region in the progression of epileptogenesis in pathological conditions. Importantly, lipid-derived endogenous cannabinoids (endocannabinoids), which are produced on demand as retrograde messengers, are central regulators of neuronal activity in the DG circuit. In this review, we summarize recent findings concerning the role of the DG in controlling hyperexcitability and propose how DG regulation by cannabinoids (CBs) could provide avenues for therapeutic interventions. We also highlight possible pathways and manipulations that could be relevant for the control of hyperexcitation. The use of CB compounds to treat epilepsies is controversial, as anecdotal evidence is not always validated by clinical trials. Recent publications shed light on the importance of the DG as a region regulating incoming hippocampal excitability during epileptogenesis. We review recent findings concerning the modulation of the hippocampal DG circuitry by CBs and discuss putative underlying pathways. A better understanding of the mechanisms by which CBs exert their action during seizures may be useful to improve therapies.     

Activation of local inhibitory circuits in the dentate gyrus by adult‐born neurons

Robust incorporation of new principal cells into pre‐existing circuitry in the adult mammalian brain is unique to the hippocampal dentate gyrus (DG). We asked if adult‐born granule cells (GCs) might act to regulate processing within the DG by modulating the substantially more abundant mature GCs. Optogenetic stimulation of a cohort of young adult‐born GCs (0 to 7 weeks post‐mitosis) revealed that these cells activate local GABAergic interneurons to evoke strong inhibitory input to mature GCs. Natural manipulation of neurogenesis by aging—to decrease it—and housing in an enriched environment—to increase it—strongly affected the levels of inhibition. We also demonstrated that elevating activity in adult‐born GCs in awake behaving animals reduced the overall number of mature GCs activated by exploration. These data suggest that inhibitory modulation of mature GCs may be an important function of adult‐born hippocampal neurons. © 2015 Wiley Periodicals, Inc.     

Neuronal gap junctions in the polymorph layer of the rat dentate gyrus

In thin sections of the rat dentate gyrus, neuronal gap junctions were observed in the polymorph layer. Gap junctions were seen on dendritic stems, on smooth and/or varicose dendrites, on spine-like appendages, and in one case on a soma. Somata of gap junction-bearing neurons showed indented nuclei with intranuclear inclusions, and received many asymmetrical and a few symmetrical synapses.     

Morphometry of hilar ectopic granule cells in the rat

Granule cell (GC) neurogenesis in the dentate gyrus (DG) does not always proceed normally. After severe seizures (e.g., status epilepticus [SE]) and some other conditions, newborn GCs appear in the hilus. Hilar ectopic GCs (EGCs) can potentially provide insight into the effects of abnormal location and seizures on GC development. Additionally, hilar EGCs that develop after SE may contribute to epileptogenesis and cognitive impairments that follow SE. Thus, it is critical to understand how EGCs differ from normal GCs. Relatively little morphometric information is available on EGCs, especially those restricted to the hilus. This study quantitatively analyzed the structural morphology of hilar EGCs from adult male rats several months after pilocarpine-induced SE, when they are considered to have chronic epilepsy. Hilar EGCs were physiologically identified in slices, intracellularly labeled, processed for light microscopic reconstruction, and compared to GC layer GCs, from both the same post-SE tissue and the NeuroMorpho database (normal GCs). Consistently, hilar EGC and GC layer GCs had similar dendritic lengths and field sizes, and identifiable apical dendrites. However, hilar EGC dendrites were topologically more complex, with more branch points and tortuous dendritic paths. Three-dimensional analysis revealed that, remarkably, hilar EGC dendrites often extended along the longitudinal DG axis, suggesting increased capacity for septotemporal integration. Axonal reconstruction demonstrated that hilar EGCs contributed to mossy fiber sprouting. This combination of preserved and aberrant morphological features, potentially supporting convergent afferent input to EGCs and broad, divergent efferent output, could help explain why the hilar EGC population could impair DG function.     

The process of reinnervation in the dentate gyrus of the adult rat: A quantitative electron microscopic analysis of terminal proliferation and reactive synaptogenesis

The present study defined the time course of terminal proliferation (the growth of presynaptic processes) and reactive Synaptogenesis in the dentate gyrus of the adult rat. Quantitative electron microscopic analyses were carried out in the dentate gyrus 2, 4, 6, 8, 10, 12, 14 days and 7 months after destruction of the ipsilateral entorhinal cortex and in the contralateral (control) dentate gyrus. At each survival interval, counts were made from photographic montages of (1) terminals (presynaptic processes with or without contacts with postsynaptic elements), (2) intact synapses, (3) degenerating synapses, (4) degeneration (degenerating presynaptic processes), and (5) multiple synapses (terminals making more than one synaptic contact). Terminal density was initially reduced to about 13% of control in the middle molecular layer at 2 and 4 days postlesion, and to about 26% of control in the outer. The density of terminals began to increase between 4 and 6 days postlesion, reaching a plateau by day 12. Synapse density was reduced to about 8% and 12% of control in the middle and outer molecular layer respectively. Synapse density increased about 5-fold between 8 and 12 days postlesion, but continued to increase in the period between 14 days and 7 months postlesion. At 2 days postlesion, the number of intact terminals that are lost corresponds to the number of degenerating presynaptic processes. This correspondence is not present at 4 days postlesion, however, suggesting a rapid removal of degenerating terminals. In contrast, even at 2 days post-lesion, the number of intact synapses that are lost does not correspond to the number of degenerating synapses. Between 2 and 10 days postlesion, the number of postsynaptic specializations is about 60% of control, but recovers slightly by 12-14 days postlesion. Qualitative and quantitative evidence suggested a collapse of spines into configurations that resembled shaft synapses. There appeared to be a deformation of degenerating presynaptic processes resulting in the appearance of multiple synapse configurations prior to reinnervation. The combined results suggest that terminal proliferation precedes reactive Synaptogenesis in the dentate gyrus by 2-4 days, that terminal proliferation is essentially complete by 12 days while reactive Synaptogenesis continues, and that multiple synapses arise at least in part as a result of a deformation of degenerating presynaptic processes rather than as a consequence of the induction of additional contacts on existing presynaptic terminals.     

GABAergic synaptic boutons in the granule cell layer of rat dentate gyrus

GABAergic synapses in the granule cell layer of the rat dentate gyrus were examined light and electron microscopically with glutamate decarboxylase (GAD) immunocytochemistry. GAD-immunoreactive synaptic boutons formed synapses with axon initial segments and somatic spines as well as somata and dendritic shafts of the granule cell. Most of these synapses were symmetrical, while a few were asymmetrical.     

Ex vivo study of dentate gyrus neurogenesis in human pharmacoresistant temporal lobe epilepsy

M. Paradisi, M. Fernández, G. Del Vecchio, G. Lizzo, G. Marucci, M. Giulioni, E. Pozzati, T. Antonelli, G. Lanzoni, G. P. Bagnara, L. Giardino and L. Calzà (2010) Neuropathology and Applied Neurobiology 36, 535–550
Ex vivo study of dentate gyrus neurogenesis in human pharmacoresistant temporal lobe epilepsy Aims: Neurogenesis in adult humans occurs in at least two areas of the brain, the subventricular zone of the telencephalon and the subgranular layer of the dentate gyrus in the hippocampal formation. We studied dentate gyrus subgranular layer neurogenesis in patients subjected to tailored antero‐mesial temporal resection including amygdalohippocampectomy due to pharmacoresistant temporal lobe epilepsy (TLE) using the in vitro neurosphere assay. Methods: Sixteen patients were enrolled in the study; mesial temporal sclerosis (MTS) was present in eight patients. Neurogenesis was investigated by ex vivo neurosphere expansion in the presence of mitogens (epidermal growth factor + basic fibroblast growth factor) and spontaneous differentiation after mitogen withdrawal. Growth factor synthesis was investigated by qRT‐PCR in neurospheres. Results: We demonstrate that in vitro proliferation of cells derived from dentate gyrus of TLE patients is dependent on disease duration. Moreover, the presence of MTS impairs proliferation. As long as in vitro proliferation occurs, neurogenesis is maintained, and cells expressing a mature neurone phenotype (TuJ1, MAP2, GAD) are spontaneously formed after mitogen withdrawal. Finally, formed neurospheres express mRNAs encoding for growth (vascular endothelial growth factor) as well as neurotrophic factors (brain‐derived neurotrophic factor, ciliary neurotrophic factor, glial‐derived neurotrophic factor, nerve growth factor). Conclusion: We demonstrated that residual neurogenesis in the subgranular layer of the dentate gyrus in TLE is dependent on diseases duration and absent in MTS.     

Morphometry of a peptidergic transmitter system: Dynorphin B‐like immunoreactivity in the rat hippocampal mossy fiber pathway before and after seizures

While the morphometry of classical transmitter systems has been extensively studied, relatively little quantitative information is available on the subcellular distribution of peptidergic dense core vesicles (DCVs) within axonal arbors and terminals, and how distribution patterns change in response to neural activity. This study used correlated quantitative light and electron microscopic immunohistochemistry to examine dynorphin B‐like immunoreactivity (dyn B‐LI) in the rat hippocampal mossy fiber pathway before and after seizures. Forty‐eight hours after seizures induced by two pentylenetetrazol injections, light microscopic dyn B‐LI was decreased dorsally and increased ventrally. Ultrastructural examination indicated that, in the hilus of the dentate gyrus, these alterations resulted from changes that were almost entirely restricted to the profiles of the large mossy‐like terminals formed by mossy fiber collaterals (which primarily contact spines), compared to the profiles of the smaller, less‐convoluted terminals found on the same collaterals (which primarily contact aspiny dendritic shafts). Dorsally, mossy terminal profile labeled DCV (lDCV) density dropped substantially, while ventrally, both mossy terminal profile perimeter and lDCV density increased. In all terminal profiles examined, lDCVs also were closely associated with the plasma membrane. Following seizures, there was a reorientation of lDCVs along the inner surface of mossy terminal profile membranes, in relation to the types of profiles adjacent to the membrane: in both the dorsal and ventral hilus, significantly fewer lDCVs were observed at sites apposed to dendrites, and significantly more were observed at sites apposed to spines. Thus, after seizures, changes specific to: (1) the dorsoventral level of the hippocampal formation, (2) the type of terminal, and (3) the type of profile in apposition to the portion of the terminal membrane examined were all observed. An explanation of these complex, interdependent alterations will probably require evoking multiple interrelated mechanisms, including selective prodynorphin synthesis, transport, and release. Hippocampus 1999; 9:255–276. © 1999 Wiley‐Liss, Inc.     

Disturbed spatial cognitive processing of body-related stimuli in a case of a lesion to the right fusiform gyrus

The fusiform gyrus (FG) is well known as one of the main neural sites of human face and body processing. We report the case of a young male patient with epilepsy and a circumscribed lesion in the right FG who presented with isolated impairments in spatial cognitive processing of body-related stimuli. However, he did not show any clinical signs of prosopagnosia. In particular, handling/processing of body and face stimuli was impaired, when stimuli were presented in unconventional views and orientations, thus requiring additional spatial cognitive operations. In this case study, we discuss the patient’s selective impairment from the view of current empirical and theoretical work on the segregation of functions in the FG.     

Reticular formation influence on neuronal transmission from perforant pathway through dentate gyrus

Electrical stimulation of the perforant pathway discharges granule cell synchronously, giving rise to a characteristic evoked potential in the granule cell layer termed here the evoked action potential or EAP. In freely moving rats, we applied 3 pulses of low intensity electrical stimulation to the medullary reticular formation prior to the application of the perforant path pulse. The effect of prior reticular formation stimulation was a marked augmentation of the normal EAP response to the perforant path stimulus. The augmentation was dependent on the behavioral state of the experimental animal (it occurred during slow-wave sleep but not during still, alert behavior) and was eliminated by anesthetic agents. The latency of EAP augmentation effect (minimum effective time interval between application of the reticular formation stimulus and the perforant path pulse) was 13--18 msec. In order to localize the sites in the medullary reticular formation from which EAP augmentation could be elicited, threshold currents for producing the effect were determined during dorso-ventral penetrations of a reticular formation stimulating electrode. EAP augmentation was elicited at low stimulus currents from a relatively broad region of the reticular formation. It was also noted that reticular formation stimulation which produced EAP augmentation always elicited one or more motor responses of the neck, back, face or vibrissae. Subsequent investigation of the pathways underlying these motor responses suggested that the effect of reticular formation stimulation on granule cell excitability was mediated by a polysynaptic pathway, the first segment of which was a projection to cells of nucleus gigantocellularis of the caudal medulla.