Bilateral Dentate Gyrus Research Articles

Electroacupuncture promotes neurogenesis in the dentate gyrus and improves pattern separation in an early Alzheimer's disease mouse model

BackgroundImpaired pattern separation occurs in the early stage of Alzheimer’s disease (AD), and hippocampal dentate gyrus (DG) neurogenesis participates in pattern separation. Here, we investigated whether spatial memory discrimination impairment can be improved by promoting the hippocampal DG granule cell neogenesis-mediated pattern separation in the early stage of AD by electroacupuncture (EA).MethodsFive familial AD mutations (5 × FAD) mice received EA treatment at Baihui and Shenting points for 4 weeks. During EA, mice were intraperitoneally injected with BrdU (50 mg/kg) twice a day. rAAV containing Wnt5a shRNA was injected into the bilateral DG region, and the viral efficiency was evaluated by detecting Wnt5a mRNA levels. Cognitive behavior tests were conducted to assess the impact of EA treatment on cognitive function. The hippocampal DG area Aβ deposition level was detected by immunohistochemistry after the intervention; The number of BrdU+/CaR+ cells and the gene expression level of calretinin (CaR) and prospero homeobox 1(Prox1) in the DG area of the hippocampus was detected to assess neurogenesis by immunofluorescence and western blotting after the intervention; The gene expression levels of FZD2, Wnt5a, DVL2, p-DVL2, CaMKII, and p-CaMKII in the Wnt signaling pathway were detected by Western blotting after the intervention.ResultsCognitive behavioral tests showed that 5 × FAD mice had impaired pattern separation (P < 0.001), which could be improved by EA (P < 0.01). Immunofluorescence and Western blot showed that the expression of Wnt5a in the hippocampus was decreased (P < 0.001), and the neurogenesis in the DG was impaired (P < 0.001) in 5 × FAD mice. EA could increase the expression level of Wnt5a (P < 0.05) and promote the neurogenesis of immature granule cells (P < 0.05) and the development of neuronal dendritic spines (P < 0.05). Interference of Wnt5a expression aggravated the damage of neurogenesis (P < 0.05), weakened the memory discrimination ability (P < 0.05), and inhibited the beneficial effect of EA (P < 0.05) in AD mice. The expression level of Wnt pathway related proteins such as FZD2, DVL2, p-DVL2, CAMKII, p-CAMKII increased after EA, but the effect of EA was inhibited after Wnt5a was knocked down. In addition, EA could reduce the deposition of Aβ plaques in the DG without any impact on Wnt5a.ConclusionEA can promote hippocampal DG immature granule cell neogenesis-mediated pattern separation to improve spatial memory discrimination impairment by regulating Wnt5a in 5 × FAD mice.

Unilateral optogenetic kindling of hippocampus leads to more severe impairments of the inhibitory signaling in the contralateral hippocampus

The kindling model has been used extensively by researchers to study the neurobiology of temporal lobe epilepsy (TLE) due to its capacity to induce intensification of seizures by the progressive recruitment of additional neuronal clusters into epileptogenic networks. We applied repetitive focal optogenetic activation of putative excitatory neurons in the dorsal CA1 area of the hippocampus of mice to investigate the role of inhibitory signaling during this process. This experimental protocol resulted in a kindling phenotype that was maintained for 2 weeks after the animals were fully kindled. As a result of the different phases of optogenetic kindling (OpK), key inhibitory signaling elements, such as KCC2 and NKCC1, exhibited distinct temporal and spatial dynamics of regulation. These alterations in protein expression were related to the distinct pattern of ictal activity propagation through the different hippocampal sublayers. Our results suggest the KCC2 disruption in the contralateral hippocampus of fully kindled animals progressively facilitated the creation of pathological pathways for seizure propagation through the hippocampal network. Upon completion of kindling, we observed animals that were restimulated after a rest period of 14-day showed, besides a persistent KCC2 downregulation, an NKCC1 upregulation in the bilateral dentate gyrus and hippocampus-wide loss of parvalbumin-positive interneurons. These alterations observed in the chronic phase of OpK suggest that the hippocampus of rekindled animals continued to undergo self-modifications during the rest period. The changes resulting from this period suggest the possibility of the development of a mirror focus on the hippocampus contralateral to the site of optical stimulations. Our results offer perspectives for preventing the recruitment and conversion of healthy neuronal networks into epileptogenic ones among patients with epilepsy.

Damage to the human dentate gyrus impairs the perceptual discrimination of complex, novel objects

The hippocampus (HPC), and the dentate gyrus (DG) subregion in particular, is purported to be a pattern separator, orthogonally representing similar information so that distinct memories may be formed. The HPC may also be involved in complex perceptual discrimination. It is unclear if this role is limited to spatial/scene stimuli or extends to the discrimination of objects. Also unclear is whether the DG itself contributes to pattern separation beyond memory. BL, an individual with bilateral DG lesions, was previously shown to have poor discrimination of similar, everyday objects in memory. Here, we demonstrate that BL's deficit extends to complex perceptual discrimination of novel objects. Specifically, BL was presented with closely matched possible and impossible objects, which give rise to fundamentally different 3D perceptual representations despite being visually similar. BL performed significantly worse than controls when asked to select an odd object (e.g., impossible) amongst three identical counterpart objects (e.g., possible) presented at different rotations. His deficit was also evident in an atypical eye fixation pattern during this task. In contrast, BL's performance was indistinguishable from that of controls on other tasks involving the same objects, indicating that he could visually differentiate the object pairs, that he perceived the objects holistically in 3D, and that he has only a mild weakness in categorizing object possibility. Furthermore, his performance on standardized neuropsychological measures indicated intact mental rotation, visual-spatial attention, and working memory (visual and auditory). Collectively, these results provide evidence that the DG is necessary for complex perceptual discrimination of novel objects, indicating that the DG might function as a generic pattern separator of a wide range of stimuli within high-level perception, and that its role is not limited to memory.

Learning to forget: Hippocampal-amygdala connectivity partially mediates the effect of sexual trauma severity on verbal recall in older women undiagnosed with posttraumatic stress disorder.

Verbal learning deficits are common among sexually traumatized women who have not been formally diagnosed with posttraumatic stress disorder (PTSD). Aberrant resting-state functional connectivity (rsFC) of the amygdala and hippocampus are implicated in PTSD and verbal memory impairment. We tested rsFC between bilateral dentate gyrus (DG) and both centromedial (CM) and basolateral (BL) nuclei of the amygdala as statistical mediators for the effect of sexual trauma-related symptom severity on delayed verbal recall performance in 63 older women (age: 60-85 years) undiagnosed with PTSD. Participant data were drawn from the NKI-Rockland Study. Individuals completed a 10-min resting-state scan, Rey Auditory Verbal Learning Test (RAVLT), and the Sexual Abuse Trauma Index (SATI) from the Trauma Symptom Checklist. Z-scores indicating rsFC of DG with BL and CM amygdala seeds were evaluated in two separate mediation models. Higher SATI scores were associated with lower RAVLT after controlling for age, β=-.23, 95% CI [.48, .03], p=.039. This effect was negated upon adding a negative path from SATI to rsFC of left DG and right CM, β=-.29, 95% CI [-.52, -.02], p=.022, and a positive path from that seed pair to RAVLT List A recall, β=.28, 95% CI [.03, 0.48], p=.015. Chi-square fit indices supported partial mediation by this seed pair, p=.762. In the absence of PTSD sexual trauma symptoms partially relate to verbal learning deficits as a function of aberrant rsFC between left hippocampus DG and right amygdala CM nuclei.

Hippocampal subfields atrophy contribute more to cognitive impairment in middle-aged patients with type 2 diabetes rather than microvascular lesions.

Neurodegeneration and microvascular lesions are related to cognitive impairment in type 2 diabetes mellitus (T2DM). We aimed to use the volume of hippocampal subfields and the burden of white matter hyperintensities (WMH) as neurodegeneration and microangiopathy markers, respectively, to investigate their potential associations with cognitive impairment in T2DM patients. A total of 76 T2DM patients and 45 neurologically unimpaired normal controls were enrolled between February 2016 to August 2018. All participants underwent structural magnetic resonance imaging (MRI) and Montreal Cognitive Assessment (MoCA). The T2DM patients were divided into the T2DM without mild cognitive impairment (T2noMCI) group (n = 44) and the T2DM with mild cognitive impairment (T2MCI) group (n = 32) according to MoCA scores. We used automatic brain segmentation and quantitative technique to assess the volume of twelve hippocampal subfields and WMH on MRI. We used age, sex, education, and total intracranial volume as our covariates and the Bonferroni method for multiple comparison correction. Both the T2MCI group and T2noMCI group showed significant hippocampal subfields atrophy compared to the controls, which were mainly in the left hippocampal tail, left CA1, bilateral molecular layer, bilateral dentate gyrus, and bilateral CA4 (all p < 0.0042). No significant differences in the volume of total WMH, deep-WMH, and periventricular-WMH were found among the three groups. The HbA1c levels were significantly negatively correlated with hippocampal atrophy, and the MoCA scores were positively correlated with bilateral hippocampal volume in T2DM patients and all samples. Mediation analyses demonstrated that the association of HbA1c levels with cognitive function was mediated by hippocampal subfields volume. Widespread hippocampal atrophies across the subfields have been found in middle-aged T2DM patients, which was positively correlated with the MoCA scores and negatively correlated with the HbA1c levels. The association of HbA1c levels with cognitive function was mediated by some crucial hippocampal subfields volume. In middle-aged patients with T2DM, the neurodegeneration is more strongly associated with cognitive impairment than microvascular lesions. Trail Registeration This study was registered on Clinical-Trails.gov (NCT02738671).

Role of Ten eleven translocation-2 (Tet2) in modulating neuronal morphology and cognition in a mouse model of Alzheimer's disease.

Abnormal expression of Ten eleven translocation-2 (Tet2) contributes to the pathogenesis of Alzheimer's disease (AD). However, to date, the role of Tet2 in modulating neuronal morphology upon amyloid-β (Aβ)-induced neurotoxicity has not been shown in a mouse model of AD. Here, we have developed a model of injured mouse hippocampal neurons induced by Aβ42 oligomers in vitro. We also investigated the role of Tet2 in injured neurons using recombinant plasmids-induced Tet2 inhibition or over-expression. We found that the reduced expression of Tet2 exacerbated neuronal damage, whereas the increased expression of Tet2 was sufficient to protect neurons against Aβ42 toxicity. Our results indicate that the brains of aged APPswe/PSEN1 double-transgenic (2×Tg-AD) mice exhibit an increase in Aβ plaque accumulation and a decrease in Tet2 expression. As a result, we have also explored the underlying mechanisms of Tet2 in cognition and amyloid load in 2×Tg-AD mice via adeno-associated virus-mediated Tet2 knockdown or over-expression. Recombinant adeno-associated virus was microinjected into bilateral dentate gyrus regions of the hippocampus of the mice. Knocking down Tet2 in young 2×Tg-AD mice resulted in the same extent of cognitive dysfunction as aged 2×Tg-AD mice. Importantly, in middle-aged 2×Tg-AD mice, knocking down Tet2 accelerated the accumulation of Aβ plaques, whereas over-expressing Tet2 alleviated amyloid burden and memory loss. Furthermore, our hippocampal RNA-seq data, from young 2×Tg-AD mice, were enriched with aberrantly expressed lncRNAs and miRNAs that are modulated by Tet2. Tet2-modulated lncRNAs (Malat1, Meg3, Sox2ot, Gm15477, Snhg1) and miRNAs (miR-764, miR-211, and miR-34a) may play a role in neuron formation. Overall, these results indicate that Tet2 may be a potential therapeutic target for repairing neuronal damage and cognitive impairment in AD.

Neuroprotective Effect of Optogenetics Varies With Distance From Channelrhodopsin-2 Expression in an Amyloid-β-Injected Mouse Model of Alzheimer's Disease.

Background: Alzheimer’s disease (AD) is a progressive neurodegenerative disease that is the most common cause of dementia. Optogenetics uses a combination of genetic engineering and light to activate or inhibit specific neurons in the brain.Objective: The objective of the study was to examine the effect of activation of glutamatergic neurons in the hippocampus of mice injected with Aβ1-42 on memory function and biomarkers of neuroinflammation and neuroprotection in the brain to elucidate the clinical utility of optogenetic neuromodulation in AD.Methods: AAV5–CaMKII–channelrhodopsin-2 (CHR2)–mCherry (Aβ-CHR2 mice) or AAV5—CaMKII–mCherry (Aβ-non-CHR2 mice) was injected into the dentate gyrus (DG) of the bilateral hippocampus of an Aβ1-42-injected mouse model of AD. The novel object recognition test was used to investigate working memory (M1), short-term memory (M2), and long-term memory (M3) after Aβ1-42 injection. Hippocampus tissues were collected for immunohistochemical analysis.Results: Compared to controls, M1 and M2 were significantly higher in Aβ-CHR2 mice, but there was no significant difference in M3; NeuN and synapsin expression were significantly increased in the DG of Aβ-CHR2 mice, but not in CA1, CA3, the subventricular zone (SVZ), or the entorhinal cortex (ENT); GluR2 and IL-10 expressions were significantly increased, and GFAP expression was significantly decreased, in CA1, CA3, the DG, and the SVZ of Aβ-CHR2 mice, but not in the ENT.Conclusion: Activation of glutamatergic neurons by optogenetics in the bilateral DG of an Aβ-injected mouse model of AD improved M1 and M2, but not M3. A single-target optogenetics strategy has spatial limitations; therefore, a multiple targeted optogenetics approach to AD therapy should be explored.

2235 15N-Leucine transport across the blood brain barrier is significantly impaired in the glutamine synthetase-inhibited brain

OBJECTIVES/SPECIFIC AIMS: Astroglial glutamine synthetase (GS), which metabolizes glutamate and ammonia to glutamine, is critical for the detoxification of brain ammonia, clearance of synaptic glutamate, and production of brain glutamine. Perturbations in the expression and activity of GS are thought to play a causative role in the pathogenesis of several conditions of abnormal neurotransmission. Although the long-term consequences of GS inhibition on amino acid homeostasis in the brain are unknown, it is thought that amino acid influx in the brain is tightly coupled with glutamine efflux via the L-type amino acid transporter. Both glutamine and leucine serve many critical functions in the brain including protein synthesis, gene expression, insulin regulation, and immune signaling. The objective of this study was to determine the effects of chronic GS inhibition with methionine sulfoximine (MSO) on glutamine and leucine homeostasis in the brain. METHODS/STUDY POPULATION: In total, 12 rats were surgically implanted with microdialysis guide cannulas in the bilateral dentate gyrus. Rats were randomly divided for surgical implantation of either a MSO (n=6) or phosphate buffer saline (PBS; n=6) pump in the right dentate gyrus. After 7 days, bilateral microdialysis probes were placed under brief isoflurane anesthesia, and microdialysis flow was established by infusing 0.5 µL/min of artificial extracellular fluid. Dialysate samples were collected every 30 minutes for the duration of the experiment. A 113 mM 15N-Leucine (3.6 mL/h) and 2 M 2–13C-sodium acetate (0.0633 μL/g/min for t=0–5 min, 0.0316 μL/g/min for t=5–10 min, and 0.0253 μL/g/min for t&gt;10 min) solution was infused intravenously for 300 minutes. The EZ:Faast Free Amino Acid analysis kit and ultra-performance liquid chromatography/tandem mass spectrometry was used for quantification of amino acids in the dialysate fluid. RESULTS/ANTICIPATED RESULTS: At baseline (t=0 h), the concentrations of glutamine were significantly lower in MSO-treated rats (p&lt;0.001) in the ipsilateral (GS-inhibited) hippocampus. There were no differences in glutamine concentrations between MSO and PBS-treated rats in the contralateral hippocampus. In PBS-treated rats, there was a significant increase in 15N-leucine between t=0 hour and t=5 hour in the contralateral (p&lt;0.05) and ipsilateral (p&lt;0.05) hippocampus. In MSO-treated rats, there was a significant increase in 15N-leucine between t=0 and t=5 hours in the contralateral (p&lt;0.05) hippocampus, but not in the ipsilateral hippocampus (p=ns.). DISCUSSION/SIGNIFICANCE OF IMPACT: This study demonstrated for the first time that basal glutamine concentrations are low in areas of the brain where GS is acutely inhibited, and that leucine uptake in these brain areas are markedly decreased. Perturbations in glutamine and leucine homeostasis have been implicated in several disease processes including diabetes, obesity, liver disease, immune system dysfunction, epilepsy, and cancer, and the glutamine-dependent leucine influx in the brain may be a novel and important therapeutic target to treat these conditions.

Altered intrinsic activity and functional connectivity in two animal models with opposing fear memories.

Post-traumatic stress disorder and sepsis-associated encephalopathy are two common neuropsychiatric disorders, but to some extent with opposing cognitive performance. However, the neural mechanisms underlying these two different phenomena remain poorly understood. In our study, the amplitude of low-frequency fluctuations (ALFF) of spontaneous blood oxygen level-dependent signals was used to assess regional disturbances in two animal models with opposing cognitive performance, namely post-traumatic stress disorder and sepsis-associated encephalopathy. Then, we examined whether the memory-relevant brain regions with impaired ALFF altered functional connectivity within the whole brain. Here, we showed that lipopolysaccharide challenge and footshock induced significantly different cognitive performance. We found significant group differences in ALFF in the bilateral dentate gyrus. In addition, our study showed that footshocks induced a significant decrease in functional connectivity between the left dentate gyrus and the visual cortex compared with the lipopolysaccharide group. Notably, cognitive performance was associated with ALFF values in the right dentate gyrus. In conclusion, our data suggested that damaged regional spontaneous activity and abnormal functional connectivity might be differently involved in two opposing fear memories.

Hypoxic Brain Injury with Bilateral Dentate Gyrus Involvement

Hypoxic Brain Injury with Bilateral Dentate Gyrus Involvement

Patterns of hippocampal neuronal loss and axon reorganization of the dentate gyrus in the mouse pilocarpine model of temporal lobe epilepsy

The patterns of hippocampal neuronal loss and rewiring of the dentate gyrus (DG) were studied in the mouse model of temporal lobe epilepsy at 2 months after pilocarpine-induced status epilepticus (PISE). NeuN immunocytochemistry showed two patterns of neuronal damage, i.e., type 1 with partial loss of pyramidal neurons in CA3 area and type 2 with almost compete loss of CA3 pyramidal neurons. Anterograde tracing with Phaseolus vulgaris leucoagglutinin (PHA-L) demonstrated that, at different rostrocaudal segments of the hippocampus, associational and commissural connections in the DG changed differently between mice with type 1 vs. type 2 neuronal loss. Calretinin (CR)-immunopositive mossy cells in ventral hilus and its fibers in inner molecular layer of bilateral DG remained in mice with type 1 but almost disappeared in mice with type 2 neuronal loss, which was further supported by retrograde labeling with cholera toxin subunit B (CTB) showing colocalization of CTB with CR in the ventral hilus of bilateral DG in mice with type 1 neuronal loss, which was lost in those with type 2 neuronal loss. Furthermore, the sprouted PHA-L-immunopositive en passant and terminal boutons from the DG were found in CA1 area to contact with surviving calbindin-, CR-, and parvalbumin-immunopositive neurons. The present study therefore suggests that different patterns of neuronal loss in CA3 area may be linked to different axon reorganizations in the DG.

Highly polysialylated neural cell adhesion molecule positive cells increased and changed localization in rat hippocampus with exposure to repeated kindled seizures

Highly polysialylated neural cell adhesion molecule (PSA-NCAM) is involved in migration of neural stem cells as well as neural plasticity. Immunoreactive PSA-NCAM expression was examined in rat with repeated exposure to amygdaloid kindled generalized seizures (GS). The number of PSA-NCAM-positive cells in bilateral dentate gyrus (DG) increased significantly at GS. Although total positive cell number was not significantly different between 3 times GS (3 GS) and 30 times GS (30 GS) groups, a greater number of positive cells were located in the outer granule cell layer (GCL), and the immunopositive dendrite greatly extended to the molecular layer in the 30 GS group. These observations indicate that increased migration of newly generated cells as well as plastic change of originally existed neural cells may occur in response to the recurrent GS, which may contribute to abnormal reconstruction of synaptic network in hippocampus and epileptogenisity in kindling.

Highly polysialylated neural cell adhesion molecule (PSA-NCAM) positive cells are increased and change localization in rat hippocampus by exposure to repeated kindled seizures.

The highly polysialylated neural cell adhesion molecule (PSA-NCAM) is involved in migration of neural stem cells as well as in neural plasticity. Immunoreactive PSA-NCAM expression was examined in rats with repeated exposure to amygdaloid kindled generalized seizures (GS). The number of PSA-NCAM positive cells in the bilateral dentate gyrus (DG) increased significantly from GS. Although the total number of positive cells was not significantly different between animals with 3 times GS (3 GS) and 30 times GS (30 GS), in the latter group a greater number of positive cells was observed in the outer granule cell layer (GCL) and a marked extension of immunopositive dendrites to the molecular layer. These observations indicate that increased migration of newly generated cells as well as plastic changes of preexisting neural cells occur in response to recurrent GS. This may contribute to an abnormal reconstruction of the synaptic network in the hippocampus and, thus, epileptogenicity from kindling.

Changes of localization of highly polysialylated neural cell adhesion molecule (PSA-NCAM) in rat hippocampus with exposure to repeated kindled seizures

Highly polysialylated neural cell adhesion molecules (PSA-NCAMs) are involved in migration of neural stem cells as well as neural plasticity. Immunoreactive PSA-NCAM expression was examined in rat with repeated exposure to amygdaloid kindled generalized seizures (GS). The number of PSA-NCAM positive cells in bilateral dentate gyrus (DG) increased significantly at GS. Although total positive cell number was not significantly different between 3 times GS (3 GS) and 30 times GS (30 GS) groups, a greater number of positive cells was located in the outer granule cell layer (GCL), and the immunopositive dendrite greatly extended to the molecular layer in the 30 GS group. These observations indicate that increased migration of newly generated cells as well as plastic change of originally-existed neural cells may occur in response to the recurrent GS, which may contribute to abnormal reconstruction of synaptic network in hippocampus and epileptogenisity in kindling.

Increment of Synapsin I Protein in the Hippocampus of Kindled Rats

Introduction: Synapsin is a vesiele‐associated protein considered to be involved in synaptogenesis and neurotransmitter release. Previously, we havc reported that synapsin I mRNA is significantly increased in the bilalcral dentate gyrus after amygdala‐kindled scizures. In this study, we investigated amygdala kindling‐induced changes i n synapsin I protein using immunohistochemistry. Quantitativc densitomctry (Mathern ct al., Ncurosciene, 1997) was performed to evaluatc changes i n synapsin I protein i n the hippocampus.Method: Fifteen inale Sprague‐Dawley rats weighing 300–350 g wcre uscd. A tripolar elcctrode was implanted stcreotaxically in the hasolateral amygdala under pcntobarbital anesthesia. After a recovery period, the rats were subjected to daily kindling stimuli (a 2‐second train of 50 HL, I‐ins rectangular waves at a currcnt intensity of 75–250 PA). Animals with implanted electrodes without atimulation were used as controls (n = 7). They werc then pcrfused 8 hours aftcr the 24–32nd (average 293th) stage 4 or 5 (Racine's classification) generalized seiztirc under anesthesia. The brains were removed and placed in 4% paraloriiialdehyde for 2 days and then transferred to 15% sucrose dissolved in 0.1 M phosphate buffer (PB) for I week. They werc then quickly frozen with dry ice powder, and coronal scctions 20 +in thick wcrc cut with a cryostat. Immunohistochemistry was performcd with polyclonal synapsin I antibody (Calhiochem). The optical density of hippocampus staining was mcasured by a computer‐bascd image analyhtem as an average of the gray value, compensated by the original control sample between white (0) and black (255). Sections were imaged using a chat‐ged‐couplcd devicc camera attached to a microscope and captured and digitized using a Macintosh computer. Illuminance waa unformly maintained and checkcd after cvery I0 measurements using the original control samplc. The image was analysed using imaging software (NIH Image). Measurements were taken at the right and left hippocampus in &gt;2 coronal sections for each animal. Results and Discussion: The kindled slice exhibited bilatcrd increases i n synapsin I protein of approximately 20%–30% in the bilateral str. lucidurn of CA3, and by approximately 20%–40% in the bilateral dentate gyrus. In the hippocampus, neuronal plastic changes including sprouting have becn shown to develop as generalized kindled scizures are repeated. Therefore, the increased synapsin I protein in these brain regions may reilect the enhanced synaptic activity and the prcsynaptic plastic changes in the specific neural network of the hippocampus of kindled rats.

Subregional analysis of amino acid levels in the guinea pig hippocampus following unilateral vestibular deafferentation

Recent behavioural and neurophysiological studies suggest that the hippocampus uses vestibular information, along with information from other sensory systems, to accomplish its spatial computational processing. By contrast, there is a dearth of neurochemical data on the interactions between the vestibular system and hippocampus. The aim of the present study was to examine levels of glutamine, glutamate, gamma-amino-butyric acid (GABA), glycine, serine, threonine and taurine in the CA1, CA2, CA3 and dentate gyrus (DG) subregions of the hippocampus at 10 hs following UVD in guinea pig, using high-performance liquid chromatography (HPLC) with electrochemical detection (ECD). Glutamine levels were not significantly different between the UVD and control conditions, except in the bilateral DG where they were significantly lower (p < 0.05). However, no significant differences in glutamate levels were observed. Although there were no significant differences in GABA levels in the DG, CA1 and CA2 regions following UVD, there was a significant increase in GABA levels in the contralateral CA3 region in the sham group compared to intact anesthetic controls. Glycine levels were significantly higher in the contralateral DG and ipsilateral CA1 in the sham groups compared to intact anesthetic controls (p < 0.05). However, in contralateral CA2 and ipsilateral CA3, glycine levels were significantly higher for both the sham and UVD groups compared to anesthetic controls (p < 0.05). Threonine levels were not significantly different in CA1; however, in ipsilateral DG and ipsilateral CA3, they were significantly higher in the sham groups compared to anesthetic controls and the UVD groups (p < 0.05). In ipsilateral CA2, threonine levels were significantly higher in the UVD group compared to anesthetic controls (p < 0.05). Taurine levels did not differ significantly in the DG, CA1 or CA3 following UVD; however, they were significantly higher in ipsilateral CA2 following UVD (p < 0.05). Finally, serine levels were not significantly different in any area of the hippocampus following UVD or sham surgery. This study provides the first data on amino acid levels in the hippocampus following UVD and sham temporal bone surgery and suggests that peripheral vestibular damage may particularly affect glutamine, glycine, threonine and taurine levels in various subregions of the hippocampus.

Increases in mRNA levels for synapsin I but not synapsin II in the hippocampus of the rat kindling model of epilepsy

We studied brain synapsin I and II mRNA levels using the amygdala kindling model of epilepsy. There were significant increases in the synapsin I mRNA level in the granule cell layer of the hippocampal bilateral dentate gyrus. One to 8 h after seizures, the level in the dentate gyrus ipsilateral to stimulation increased by 44.2-73.2%, compared with the control level. Of the time points investigated, the greatest increase in expression was observed 8 h after the kindled seizures. Furthermore, the synapsin I mRNA levels in the dentate gyrus contralateral to stimulation increased by 28.0% and 51.1%, 2 and 8 h, respectively, after the kindled seizures. Expression of this mRNA, however, did not change significantly in other areas examined, including CA1, CA2, CA3 and the polymorphic layer of the hippocampus and the perirhinal and temporal cortices. Synapsin II mRNA levels did not change significantly in any of the regions studied for up to 24 h after the seizures and synapsin II was presumed to have little involvement in kindling. We considered the locally elevated synapsin I mRNA levels in the bilateral dentate gyrus associated with kindling indicate that excitatory changes occur in the synaptic circuit in which the dentate granule cells participate. Synapsin I may be involved in the presynaptic molecular mechanisms underlying the neuronal plasticity in kindling.

Increased Levels of mRNA for β- but not α-Subunit of Calmoduline Kinase II Following Kindled Seizures

We studied levels of mRNA for the α- and β-subunits of calmodulin (CaM) kinase II using the amygdaloid kindling model of epilepsy. There were significant increases in mRNA for the β-subunit of CaM kinase II in the hippocampus 4—24 h after stage 5-kindled seizures. Moreover, this mRNA was significantly increased by 20.0–26.5% in the bilateral dentate gyrus 8 to 24 h after kindled seizures. The β-subunit mRNA was also significantly increased by 13.5–19.0% in the CA3 on the side ipsilateral to the stimulation, 4 to 8 h after kindled seizures. mRNA for the α-subunit of CaM kinase II was not significantly changed in the regions examined for up to 24 h after the kindled seizures. These results suggest that CaM kinase II mediates the molecular processes that follow kindled seizures. It is possible that increases in CaM kinase II-dependent protein phosphorylation are associated with the plastic changes in kindling.

Associative synaptic potentiation and depression: Quantification of dissociable modifications in the hippocampal dentate gyrus favors a particular class of synaptic modification equations

This report further characterizes associative long-term synaptic modification of the ipsilateral and contralateral synapses formed by the bilateral entorhinal cortical (EC) projection to the dentate gyrus (DG). The experimental model is the anesthetized hooded rat. The quantitative results qualify this system as a model for studying the rules of associative synaptic modification formulated in terms of individual synapses. Bilateral DG microelectrodes recorded both ipsilateral and contralateral EC-DG responses before and after brief, high-frequency EC conditioning stimulation. The weak contralateral pathway received high-frequency conditioning before, during, or after similar conditioning of the strong, converging ipsilateral pathway. Statistical analyses revealed two types of significant, dissociated synaptic modifications, which depend on the relationship of the ipsilateral and contralateral afferents. First, contralateral EC-DG responses potentiated or depressed when the converging ipsilateral responses concurrently either potentiated or remained unchanged. Second, contralateral EC-DG responses potentiated, depressed, or showed no change when the collateral ipsilateral responses concurrently either potentiated or remained unchanged. Correlation and contingency table analyses indicated that changes in the contralateral synaptic responses are not well predicted by changes at either neighboring synapses of the converging ipsilateral pathway or at synapses of the collateral ipsilateral pathway. The contingencies of associated pre- and postsynaptic activation determined by the conditioning paradigm, however, accurately predicted the altered synaptic responses of both ipsilateral and contralateral EC-DG pathways. The results imply that associative synaptic modification in the EC-DG system is specific to individual synapses and requires both appropriate presynaptic and postsynaptic activation. Because this system provides suitable controls for nonspecific effects of conditioning stimulation and because modification of neighboring synapses is dissociable, the EC-DG system can be used to study further those rules of activity-dependent associative modification that are formulated in terms of individual synapses. The discussion briefly considers published rules of synaptic modification, pointing out several rules that are not consistent with the experimental observations and one that agrees with the present results.