Swimming exercise improves short- and long-term memories: Time-course changes.

The beneficial effects of exercise training on memory formation are well documented. However, the memory enhancement profile following the time-course of exercise training remains unknown. In this investigation, changes in the spatial hippocampal memory following a time-course of swimming exercise training were examined. Young adult Wistar rats were tested for both short-term and long-term memories using the radial arm water maize (RAWM) paradigm following 0, 1, 7, 14, and 28 days of swimming exercise training (60 min per day, 5 days/week)s. The mean total errors on RAWM during the learning phase and memory testing remained the same (p > 0.5) after 1 day of swimming exercise. On the other hand, swimming exercise-induced significant enhancement to the learning phase and memory formation after 7 days of training (p < 0.01). Errors decreased (p < 0.0001) after 7 days of training and remained lower (p < 0.0001) than baseline without differences between 7, 14, and 28 days (p > 0.5). Similarly, short- and long-term memories improved after 7 days (p < 0.05) of training as compared to the baseline without differences between 7, 14, and 28 days (p > 0.05). The time course of improvement of learning and both short- and long-term memories after swimming exercise were evident after 7 days and plateaued thereafter. Results of the current study could form the base for future utilization of exercises to enhance cognitive function in healthy individuals.

Protective Effects of Aquaporin-4 Deficiency on Longer-term Neurological Outcomes in a Mouse Model.

Traumatic brain injury (TBI) has been a crucial health problem, with more than 50 million patients worldwide each year. Glymphatic system is a fluid exchange system that relies on the polarized water channel aquaporin-4 (AQP4) at the astrocytes, accounting for the clearance of abnormal proteins and metabolites from brain tissues. However, the dysfunction of glymphatic system and alteration of AQP4 polarization during the progression of TBI remain unclear. AQP4-/- and Wild Type (WT) mice were used to establish the TBI mouse model respectively. Brain edema and Evans blue extravasation were conducted 24 h post-injury to evaluate the acute TBI. Morris water maze (MWM) was used to establish the long-term cognitive functions of AQP4-/- and WT mice post TBI. Western-blot and qRT-PCR assays were performed to demonstrate protective effects of AQP4 deficiency to blood-brain barrier (BBB) integrity and amyloid-ß clearance. The inflammation of cerebral tissues post TBI was estimated by ELISA assay. AQP4 deficiency alleviated the brain edema and neurological deficit in TBI mice. AQP4-knockout led to improved cognitive outcomes in mice post TBI. The BBB integrity and cerebral amyloid-ß clearance were protected by AQP4 deficiency in TBI mice. AQP4 deficiency ameliorated the TBI-induced inflammation. AQP4 deficiency improved longer-term neurological outcomes in a mouse model of TBI.

Chronic Cyanuric Acid Exposure Depresses Hippocampal LTP but Does Not Disrupt Spatial Learning or Memory in the Morris Water Maze.

Exposure to cyanuric acid (CA) causes multiple organ failure accompanied by the involvement in kinds of target proteins, which are detectable and play central roles in the CNS. The hippocampus has been identified as a brain area which was especially vulnerable in developmental condition associated with cognitive dysfunction. No studies have examined the effects of CA on hippocampal function after in vitro or in vivo treatment. Here, we aimed to examine hippocampal synaptic function and adverse behavioral effects using a rat model administered CA intraperitoneally or intrahippocampally. We found that infusion of CA induced a depression in the frequency but not the amplitude of spontaneous excitatory postsynaptic currents (sEPSCs), miniature excitatory postsynaptic currents (mEPSCs), or N-methyl-D-aspartate (NMDA)-mediated excitatory postsynaptic currents (EPSCs) of the CA1 neurons in dose-dependent pattern. Both intraperitoneal and intrahippocampal injections of CA suppressed hippocampal LTP from Schaffer collaterals to CA1 regions. Paired-pulse facilitation (PPF), a presynaptic phenomenon, was enhanced while the total and phosphorylated expression of NMDA-GluN1, NMDA-GluN2A, and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-GluA1 subunits were comparable between CA-treated and control groups. In Morris water maze test, both groups could effectively learn and retain spatial memory. Our studies provide the first evidence for the neurotoxic effect of CA and the insight into its potential mechanisms.

Sex differences in the IntelliCage and the Morris water maze in the APP/PS1 mouse model of amyloidosis.

Transgenic rodent models were created to decipher pathogenic mechanisms associated with Alzheimer's disease (AD), and behavioral apparatuses such as the Morris water maze (MWM) are used to assess cognition in mice. The IntelliCage was designed to circumvent issues of traditional behavioral tests, such as frequent human handling. The motivation to complete IntelliCage tasks is water consumption, which is less stressful than escaping from a pool in the MWM. Here, we examined behavioral performances of mice in the IntelliCage and MWM tasks. Twelve-month-old male and female APP/PS1 and non-transgenic mice first underwent 42 days of IntelliCage testing to assess prefrontal cortical and hippocampal function followed by MWM testing for six days. We found that females performed better in the IntelliCage while males performed superiorly in the MWM. Mechanistically, female APP/PS1 mice had a higher Amyloid-ß plaque load throughout the brain, which is inconsistent with their performance in the IntelliCage. Collectively, these results inform scientists about the sex-based differences when testing animals in different behavioral paradigms that tap similar cognitive functions.

Hippocampal Aromatase Knockdown Aggravates Ovariectomy-Induced Spatial Memory Impairment, Aß Accumulation and Neural Plasticity Deficiency in Adult Female Mice.

Ovarian estrogens (mainly 17ß estradiol, E2) have been involved in the regulation of the structure of hippocampus, the center of spatial memory. In recent years, high levels of aromatase (AROM), the estrogen synthase, has been localized in hippocampus; and this hippocampus-derived E2 seems to be functional in synaptic plasticity and spatial memory as ovarian E2 does. However, the contribution of ovarian E2 and hippocampal E2 to spatial memory and neural plasticity remains unclear. In this study, AROM-specific RNA interference AAVs (shAROM) were constructed and injected into the hippocampus of control or ovariectomized (OVX) mice. Four weeks later the spatial learning and memory behavior was examined with Morris water maze, the expression of hippocampal Aß related proteins, selected synaptic proteins and CA1 synapse density, actin polymerization related proteins and CA1 spine density were also examined. The results showed that while OVX and hippocampal shAROM contributed similarly to most of the parameters examined, shAROM induced more increase in BACE1 (amyloidogenic ß-secretase), more decrease in neprilysin (Aß remover) and Profilin-1 (actin polymerization inducer). More importantly, combined OVX and shAROM treatment displayed most significant impairment of spatial learning and memory as well as decrease in synaptic plasticity compared to OVX or shAROM alone. In conclusion, the above results clearly demonstrated the crucial role of hippocampal E2 in the regulation of the structure and function of hippocampus besides ovarian E2, indicating that hippocampal E2 content should also be taken into consideration during estrogenic replacement.

Prenatal Hypoxia Induces Premature Aging Accompanied by Impaired Function of the Glutamatergic System in Rat Hippocampus.

Prenatal hypoxia is among leading causes of progressive brain pathologies in postnatal life. This study aimed to analyze the characteristics of the hippocampal glutamatergic system and behavior of rats in early (2 weeks), adult (3 months) and advanced (18 months) postnatal ontogenesis after exposure to prenatal severe hypoxia (PSH, 180 Torr, 5% O2, 3 h) during the critical period in the formation of the hippocampus (days 14-16 of gestation). We have shown an age-dependent progressive decrease in the hippocampal glutamate levels, a decrease of the neuronal cell number in the CA1 hippocampal region, as well as impairment of spatial long-term memory in the Morris water navigation task. The gradual decrease of glutamate was accompanied by decreased expression of the genes that mediate glutamate metabolism and recycling in the hippocampus. That deficiency apparently correlated with an increase of the metabotropic glutamate receptor type 1 (mGluR1) and synaptophysin expression. Generation of the lipid peroxidation products in the hippocampus of adult rats subjected to prenatal severe hypoxia (PSH rats) was not increased compared to the control animals when tested in a model of glutamate excitotoxicity induced by severe hypoxia. This demonstrates that excessive glutamate sensitivity in PSH rats does not compensate for glutamate deficiency. Our results show a significant contribution of the glutamate system dysfunction to age-associated decrease of this mediator, cognitive decline, and early neuronal loss in PSH rats.

Drinking water temperature affects cognitive function and progression of Alzheimer's disease in a mouse model.

Lifestyle factors may affect mental health and play a critical role in the development of neurodegenerative diseases including Alzheimer's disease (AD). However, whether the temperatures of daily beverages have any impact on cognitive function and AD development has never been studied. In this study, we investigated the effects of daily drinking water temperatures on cognitive function and AD development and progression in mice and the underlying mechanisms. Cognitive function of mice was assessed using passive avoidance test, open field test, and Morris water maze. Wild-type Kunming mice receiving intragastric water (IW, 10 mL/kg, 2 times/day) at 0 °C for consecutive 15 days displayed significant cognitive defects accompanied by significant decrease in gain of body weight, gastric emptying rate, pepsin activity, and an increase in the energy charge in the cortex when compared with mice receiving the same amount of IW at 25 °C (a temperature mimicking most common drinking habits in human), suggesting the altered neuroenergetics may cause cognitive decline. Similarly, in the transgenic APPwse/PS1De9 familial AD mice and their age- and gender-matched wild-type C57BL/6 mice, receiving IW at 0 °C, but not at 25 °C, for 35 days caused a significant time-dependent decrease in body weight and cognitive function, accompanied by a decreased expression of PI3K, Akt, the glutamate/GABA ratio, as well as neuropathy with significant amyloid lesion in the cortex and hippocampus. All of these changes were significantly aggravated in the APPwse/PS1De9 mice than in the control C57BL/6 mice. These data demonstrate that daily beverage at 0 °C may alter brain insulin-mediated neuroenergetics, glutamate/GABA ratio, cause cognitive decline and neuropathy, and promote AD progression.

Accelerated Failure Time Survival Model to Analyze Morris Water Maze Latency Data.

Traumatic brain injury (TBI) induces cognitive deficits clinically and in animal models. Learning and memory testing is critical when evaluating potential therapeutic strategies and treatments to manage the effects of TBI. We evaluated three data analysis methods for the Morris water maze (MWM), a learning and memory assessment widely used in the neurotrauma field, to determine which statistical tool is optimal for MWM data. Hidden platform spatial MWM data aggregated from three separate experiments from the same laboratory were analyzed using 1) a logistic regression model, 2) an analysis of variance (ANOVA) model, and 3) an accelerated failure time (AFT) time-to-event model. The logistic regression model showed no significant evidence of differences between treatments among any swims over all days of the study, p > 0.11. Although the ANOVA model found significant evidence of differences between sham and TBI groups on three out of four swims on the third day, results are potentially biased due to the failure of this model to account for censoring. The time-to-event AFT model showed significant differences between sham and TBI over all swims on the third day, p < 0.045, taking censoring into account. We suggest AFT models should be the preferred analytical methodology for latency to platform associated with MWM studies.

High-Frequency rTMS Improves Cognitive Function by Regulating Synaptic Plasticity in Cerebral Ischemic Rats.

Poststroke cognitive impairment (PSCI) is one of the most severe sequelae of stroke and lacks effective treatment. Previous studies have shown that high-frequency repetitive transcranial magnetic stimulation (rTMS) may be a promising PSCI therapeutic approach, but the underlying mechanism is unclear. To uncover the effect of rTMS on PSCI, a transient middle cerebral artery occlusion (tMCAO) model was established. Modified Neurological Severity Score (mNSS) test and Morris Water Maze (MWM) test were performed to assess the neurological and cognitive function of rats. Furthermore, to explore the underlying mechanism, differentially expressed genes (DEGs) in the hippocampus of rats in the rTMS group and tMCAO group were compared using RNA sequencing. Then, bioinformatics analysis, including gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and protein-protein interaction (PPI) network analysis, was conducted to elaborate these DEGs. Our results indicated that high-frequency rTMS could significantly improve neurological and cognitive function, according to mNSS and MWM tests. We found 85 DEGs, including 71 upregulated genes and 14 downregulated genes, between the rTMS group and tMCAO group. The major functional category was related to chemical synaptic transmission modulation and several DEGs were significantly upregulated in processes related to synaptic plasticity, such as glutamatergic synapses. Calb2, Zic1, Kcnk9, and Grin3a were notable in PPI analysis. These results demonstrate that rTMS has a beneficial effect on PSCI, and its mechanism may be related to the regulation of synaptic plasticity and functional genes such as Calb2, Zic1, Kcnk9, and Grin3a in the hippocampus.

The modulation of the spatial reference memory by the orexinergic system of the dorsal raphe nucleus.

The dorsal raphe nucleus (DRN) is a brainstem nucleus involved in the pathophysiology of the depression, through its serotoninergic innervation. Furthermore, depressive symptoms in patients are also associated with some memory and sleep complaints. Anatomical evidence confirmed the presence of projections from the lateral hypothalamus to serotonergic neurons of the dorsal raphe nucleus (DRN). These projection fibers release orexin neuropeptides which play roles in the spatial memory. Both of the orexinergic receptors are widely distributed in dorsal raphe nucleus. Therefore, the present work was aimed to assess the probable roles of orexin 1 and 2 receptors using an orexin 1 receptor antagonist, SB-334867-A, and an orexin 2 receptor antagonist, TCS-OX2-29 in the DRN on the retrieval, and consolidation phases of spatial reference memory in the Morris water maze (MWM) task. The results demonstrated that blocking orexin 1 receptors in the DRN impairs the process of memory consolidation in the spatial MWM via increasing in the time of the escape latency of the probe day. Blocking these receptors did not affect the retrieval phase of MWM learning. Furthermore, blocking of the orexin 2 receptors in this area did not affect neither consolidation nor retrieval phases of the memory. In conclusion, orexin 1 receptors in the DRN play major roles in the consolidation of the spatial reference memory in rats.

Bidirectional regulation of distinct memory domains by α5-subunit-containing GABAA receptors in CA1 pyramidal neurons.

Reduction in the expression or function of α5-subunit-containing GABAA receptors (α5GABAARs) leads to improvement in several hippocampus-dependent memory domains. However, studies thus far mostly lack anatomical specificity in terms of neuronal circuits and populations. We demonstrate that mice with a selective knockdown of α5GABAARs in CA1 pyramidal neurons (α5CA1KO mice) show improved spatial and trace fear-conditioning memory. Unexpectedly, α5CA1KO mice were comparable to controls in contextual fear-conditioning but showed an impairment in context discrimination, suggesting fine-tuning of activity in CA1 pyramidal cell dendrites through α5-mediated inhibition might be necessary for distinguishing highly similar contexts.

Enriched Environment and Social Isolation Affect Cognition Ability via Altering Excitatory and Inhibitory Synaptic Density in Mice Hippocampus.

The purpose of the study was to examine whether the underlying mechanism of the alteration of cognitive ability and synaptic plasticity induced by the housing environment is associated with the balance of excitatory/inhibitory synaptic density. Enriched environment (EE) and social isolation (SI) are two different housing environment, and one is to give multiple sensory environments, the other is to give monotonous and lonely environment. Male 4-week-old C57 mice were divided into three groups: CON, EE and SI. They were housed in the different cage until 3 months of age. Morris water maze and novel object recognition were performed. Long term potentiation (LTP), depotentiation (DEP) and local field potentials were recorded in the hippocampal perforant pathway and dentate gyrus (DG) region. The data showed that EE enhanced the ability of spatial learning, reversal learning and memory as well as LTP/DEP in the hippocampal DG region. Meanwhile, SI reduced those abilities and the level of LTP/DEP. Moreover, there were higher couplings of both phase-amplitude and phase-phase in the EE group, and lower couplings of them in the SI group compared to that in the CON group. Western blot and immunofluorescence analysis showed that EE significantly enhanced the level of PSD-95, NR2B and DCX; however, SI reduced them but increased GABAARα1 and decreased DCX levels. The data suggests that the cognitive functions, synaptic plasticity, neurogenesis and neuronal oscillatory patterns were significantly impacted by housing environment via possibly changing the balance of excitatory and inhibitory synaptic density.

Ganoderma lucidum aqueous extract prevents hypobaric hypoxia induced memory deficit by modulating neurotransmission, neuroplasticity and maintaining redox homeostasis.

Oxidative stress due to hypobaric hypoxia at extreme altitudes causes severe neuronal damage and irreversible cognitive loss. Owing to contraindications of current drug therapies, the aim of the study was to investigate memory enhancing potential of aqueous extract of Ganoderma lucidum (GLAQ) and underlying neuroprotective mechanism using rat hypobaric hypoxia test model. Rats exposed to hypobaric hypoxia showed deranged spatial memory in morris water maze test with hippocampal damage and vasogenic cerebral edema. All these changes were prevented with GLAQ treatment. Blood and biochemical analysis revealed activation of hypoxic ventilatory response, red blood cells induction, reversal of electrolyte and redox imbalance, and restoration of cellular bioenergetic losses in GLAQ treated animals. Notably, GLAQ treatment ameliorated levels of neurotransmitters (catecholamines, serotonin, glutamate), prevented glucocorticoid and α-synuclein surge, improved neuroplasticity by upregulating CREB/p-CREB/BDNF expression via ERK1/ERK2 induction. Further, restoration of nuclear factor erythroid 2-related factor with stabilization of hypoxia inducible factors and inflammatory markers were evidenced in GLAQ treated rats which was additionally established in gene reporter array using an alternative HT22 cell test model. Conclusively, our studies provide novel insights into systemic to molecular level protective mechanism by GLAQ in combating hypobaric hypoxia induced oxidative stress and memory impairment.

Amyloid Fibril-Induced Astrocytic Glutamate Transporter Disruption Contributes to Complement C1q-Mediated Microglial Pruning of Glutamatergic Synapses.

The complement C1q plays a critical role in microglial phagocytosis of glutamatergic synapses and in the pathogenesis of neuroinflammation in Alzheimer's disease (AD). We recently reported that upregulation of metabotropic glutamate receptor signaling is associated with increased synaptic C1q production and subsequent microglial phagocytosis of synapses in the rodent models of AD. Here, we explored the role of astrocytic glutamate transporter in the synaptic C1q production and microglial phagocytosis of hippocampal glutamatergic synapses in a rat model of AD. Activation of astrocyte and reduction glutamate transporter 1 (GLT1) were noted after bilateral microinjection of amyloid-beta (Aß1-40) fibrils into the hippocampal CA1 area of rats. Ceftriaxone is a ß-lactam antibiotic that upregulates GLT1 expression. Bilateral microinjection of ceftriaxone recovered GLT1 expression, decreased synaptic C1q production, suppressed microglial phagocytosis of glutamatergic synapses in the hippocampal CA1, and attenuated synaptic and cognitive deficits in rats microinjected with Aß1-40. In contrast, artificial suppression of GLT1 activity by DL-threo-beta-benzyloxyaspartate (DL-TBOA) in naïve rats induced synaptic C1q expression and microglial phagocytosis of glutamatergic synapses in the hippocampal CA1 area, resulting in synaptic and cognitive dysfunction. These findings demonstrated that impairment of astrocytic glutamate transporter plays a role in the pathogenesis of AD.

Oligodendrocytes Death Induced Sensorimotor and Cognitive Deficit in N-nitro-L-arginine methyl Rat Model of Pre-eclampsia.

Pre-eclampsia (PE) is a pregnancy complicated syndrome that affects multiple organs including the brain that continue post- delivery in both mother and the offspring. We evaluated the expression of oligodendrocytes in the brain of PE rat model through development as well as the cognitive changes and other behavioural modifications that may occur later in the life of offspring of PE-like rat model. Pregnant rats divided into early-onset and late-onset groups were administered with N-nitro- L-arginine methyl (L-NAME) through drinking water at gestational days (GD) 8-17. Rats were allowed free access to water throughout the pregnancy. At GD 19, post-natal day (PND) 1 and 60, rats were sacrificed and brain excised for further analysis. The offspring were subjected to behavioural studies for cognitive and sensorimotor impairments before sacrificed at PND 60. Results showed significant down-regulation in the expression of OLIG2 in PE at GD 19 brain which persists till PND 60. Likewise, there was a significant increase in the latency to locate the platform in Morris water maze, time to traverse the balance beam and reduced hanging time on the wire test between the control and the PE treated. PE could lead to impaired neuronal signalling through demyelination which may contributes significantly to long-term sensorimotor and cognitive deficit.

Morris water maze overtraining increases the density of thorny excrescences in the basal dendrites of CA3 pyramidal neurons.

The hippocampus plays a fundamental role in spatial learning and memory. Dentate gyrus (DG) granular neurons project mainly to proximal apical dendrites of neurons in the CA3 stratum lucidum and also, to some extent, to the basal dendrites of CA3 pyramidal cells in the stratum oriens. The terminal specializations of DG neurons are the mossy fibers (MF), and these huge axon terminals show expansion in the CA3 stratum oriens after the animals undergo overtraining in the Morris Water Maze task (MWM). However, to our knowledge there are no reports regarding the possible changes in density of post-synaptic targets of these terminals in the basal dendrites of CA3 neurons after overtraining in the MWM. The purpose of this work was to study the density of thorny excrescences (TE) and other dendritic spine types (stubby, thin, and mushroom) in the CA3 stratum oriens in animals overtrained in the MWM for three consecutive days and in animals trained for only one day. Seven days after MWM training, the animals were sacrificed, and their brains removed and processed for rapid Golgi staining to visualize the different types of dendritic protrusions. Our results revealed that the relative quantity of stubby, thin, and mushroom dendritic spines did not change, regardless of amount of training. However, a significant increase in the density of TE was detected in the overtrained animals. These results strongly suggest that spatial water maze overtraining induces an increased density of MF-TE connections, which might be functionally relevant for long-term spatial memory formation.