Gastrodin Ameliorates Memory Deficits in 3,3′-Iminodipropionitrile-Induced Rats: Possible Involvement of Dopaminergic System

3,3′-Iminodipropionitrile (IDPN), one of the nitrile derivatives, can induce neurotoxicity, and therefore cause motor dysfunction and cognitive deficits. Gastrodin is a main bioactive constituent of a Chinese herbal medicine (Gastrodia elata Blume) widely used for treating various neurological disorders and showed greatly improved mental function. This study was designed to determine whether administration of gastrodin attenuates IDPN-induced working memory deficits in Y-maze task, and to explore the underlying mechanisms. Results showed that exposure to IDPN (150 mg/kg/day, v.o.) significantly impaired working memory and that long-term gastrodin (200 mg/kg/day, v.o.) could effectively rescue these IDPN-induced memory impairments as indicated by increased spontaneous alternation in the Y-maze test. Additionally, gastrodin treatment prevented IDPN-induced reductions of dopamine (DA) and its metabolites, as well as elevation of dopamine turnover ratio (DOPAC + HVA)/DA. Gastrodin treatment also prevented alterations in dopamine D2 receptor and dopamine transporter protein levels in the rat hippocampus. Our results suggest that long-term gastrodin treatment may have potential therapeutic values for IDPN-induced cognitive impairments, which was mediated, in part, by normalizing the dopaminergic system.     

Protective Effect of Silibinin on Learning and Memory Impairment in LPS-Treated Rats via ROS–BDNF–TrkB Pathway

Silibinin, a flavonoid derived from the herb milk thistle (Silybum marianum), has been used as a hepato-protectant in the clinical treatment of liver disease. In the present study, the effect of silibinin on lipopolysaccharide (LPS)-induced neuroinflammatory impairment in rats is investigated. Injection of LPS into lateral ventricle caused learning and memory impairment. Rats were treated with silibinin to see the effect in comparison with resveratrol as a positive control. Y-maze and Morris water maze tests showed that silibinin significantly attenuated memory damage caused by LPS treatment. At the molecular analysis, the levels of IL-1β and of IL-4 in the hippocampus were decreased and enhanced, respectively, by the treatment with silibinin. NF-κB expression was attenuated by silibinin treatment. Furthermore, generation of total reactive oxygen species (ROS) in the hippocampus was elevated in silibinin-treated groups, and so were the expressions of brain-derived neurotrophic factor (BDNF) and tyrosine receptor kinase B (TrkB). At the same time, LPS-induced reduction of neurons in hippocampus was reversed by silibinin. In conclusion, silibinin ameliorated the impairment of learning and memory of LPS-injection rats, possibly due to the activation of ROS–BDNF–TrkB pathway in the hippocampus as well as the suppression of inflammatory response. This study gives an insight on the beneficial consequences of ROS in central nervous system. Silibinin might be a potential candidate drug for neurodegenerative diseases.     

Struvite Urolithiasis in Long–Evans Rats

Struvite urinary calculi, which are composed of magnesium, ammonium, and phosphate, can cause complications including sepsis and renal failure. Struvite calculi were identified within the urinary bladder and renal pelvis of 2 Long-Evans rats that died within days after arrival from a commercial vendor. The remaining rats in the shipment were screened by physical examination, radiography, and ultrasonography, revealing an additional 2 animals that were clinically affected. These rats were euthanized, necropsied, and yielded similar findings to those from the first 2 rats. In addition, urine samples had an alkaline pH and contained numerous bacteria (predominantly Proteus mirabilis), leukocytes, and crystals. All calculi were composed completely of struvite. Another 7 rats in the shipment had alkaline urine with the presence of blood cells; 6 of these rats also had abundant struvite crystals, and P. mirabilis was cultured from the urine of 3 rats. Further investigation by the vendor identified 2 of 100 rats with struvite calculi from the same colony. Although no specific cause could be implicated, the fact that all the affected rats came from the same breeding area suggests a genetic or environmental triggering event; a contribution due to diet cannot be ruled out. Our findings suggest that the affected rats had metabolic disturbances coupled with bacterial infection that predisposed them to develop struvite calculi. During sudden increases of struvite urinary calculi cases in rats, urine cultures followed by appropriate surgical intervention and antibiotic therapy is warranted. Additional factors, including diet, merit attention as well.Struvite, also known as triple phosphate, is a crystalline substance composed of magnesium ammonium phosphate (MgNH₄PO₄• 6 H₂0)¹⁰ that was first identified in the 18th century. This mineral is what consolidates into urinary calculi, or stones, both in humans and animals. Other frequently encountered types of urinary stones include calcium oxalate, calcium phosphate, uric acid, and cysteine.¹⁷ Although only 2% to 3% of stones from humans are composed of struvite,⁴³ struvite calculi are important clinically because they can lead to sepsis and renal failure.⁴³In both humans and animals, stones in the urinary tract can obstruct the urine outflow, with subsequent extreme pain, hydronephrosis, and (possibly) rupture of the urinary bladder. In addition, disruption of kidney function can lead to metabolic imbalances, such as uremia, seizures, depression, anorexia, dehydration, even coma and death.^1,8 Here we present a case study describing the spontaneous presentation of struvite urolithiasis in a recently imported cohort of rats and related significant clinical findings.     

Impaired Spatial Learning and Memory after Sevoflurane–Nitrous Oxide Anesthesia in Aged Rats Is Associated with Down-Regulated cAMP/CREB Signaling

Neurocognitive deficits arising from anesthetic exposure have recently been debated, while studies have shown that the phosphorylation of cyclic AMP response element-binding protein (CREB) in the hippocampus is critical for long-term memory. To better understand the neural effects of inhalational anesthetics, we studied the behavioral and biochemical changes in aged rats that were exposed to sevoflurane (Sev) and nitrous oxide (N₂O) for 4 h. Eighteen-month-old rats were randomly assigned to receive 1.3% sevoflurane and 50% nitrous oxide/50% oxygen or 50% oxygen for 4 h. Spatial learning and memory were tested with the Morris water maze 48 h after exposure, and the results showed that sevoflurane–nitrous oxide exposure induced a significant deficit in spatial learning acquisition and memory retention. Experiments revealed that the cAMP and pCREB levels in the dorsal hippocampus were decreased in rats with anesthetic exposure in comparison with control rats 48 h after anesthesia as well as 15 min after the probe trial, but there were no significant differences in CREB expression. Besides these, the current study also found the DG neurogenesis significantly decreased as well as neuronal loss and neuronal apoptosis increased in the hippocampus of rats exposed to Sev+N₂O. The current study demonstrated that down-regulation of cAMP/CREB signaling, decrease of CREB-dependent neurogenesis and neuronal survival in the hippocampus contributed to the neurotoxicity and cognitive dysfunction induced by general anesthesia with sevoflurane–nitrous oxide.     

Study on Embryo-Fetal Development to Artesunate in Rats

The present study was conductedtoinvestigate the potential embryo-fetal toxicity of Artesunate,an antimalarial drug,inSprague-Dawleyrats.Thetestitemwas orallyadministered bygavageto pregnant rats(22females per group)fromgesta-tional day(GD)6through15at doselevels of0,2,4and8mg/kg,All dams were subjectedtocaesareansectionon GD20andtheir fetuses were examined for external,visceral,and skeletal abnormalities.There were no treatment-relatedclinical signs,body weights andfood consumptionin any o…     

Memory Deficit in Mice Administered Aluminum–maltolate Complex

Recently, aluminum (Al) has been identified as one of the environmental factors responsible for cause certain nerve degeneration diseases, particularly, Alzheimer’s disease (AD). However, the relationship between Al and AD is controversial. We previously examined whether Al induced neurotoxin in the brain of mice when aluminum–maltolate complex (ALM) was administered daily for 120 days. Our results revealed that Al accumulated in the brain induced oxidative stress, and the nerve degeneration was detected in the brain of the ALM-treated group. On the basis of these results, we have tried to examine whether the incorporated Al affects memory in mice with regard to an indicator of spatial memory deficits depending on the chemical forms of Al, namely, as an ion (AlCl₃) and in the form of a complex (ALM). We administered saline, AlCl₃, and ALM at a concentration of 40 μmol Al/kg body weight to mice by daily ip injections for 60 days. We assessed spatial memory by a water maze task and determined the Al levels in the brain of the mice by the neutron activation analysis method. Spatial memory deficit as an indicator of the swimming time was related to Al accumulation in the brain of mice; the chemical form of the Al compound was important in order to exhibit the memory deficit in mice; the uptake of Al is higher in mice when it is administered in a complex form than in an ionic form.     

Intranigral Dopamine Toxicity and α-Synuclein Response in Rats

There is increasing evidence that, in addition to its function as the main neurotransmitter in the nigrostriatal pathway, dopamine (DA) may be neurotoxic in certain conditions. In this study, the toxicity of DA was assessed by direct injection into the substantia nigra of anaesthetised rats, and its effects were compared with those of 6-hydroxydopamine. Brains were removed 1, 2 and 3 weeks after the lesion for histological or neurochemical analysis. DA caused a significant loss of 35% of tyrosine hydroxylase-positive neurons in the pars compacta of substantia nigra and a 40% reduction of striatal DA content. Cells with signs compatible with both apoptosis and autophagy were observed. GADD153, a parameter of endoplasmic reticulum stress, was strongly induced by 6-hydroxydopamine but not by DA. DA increased the α-synuclein content 1 week after the lesion (but not at the later times analyzed) in tyrosine hydroxylase-positive and in non-dopaminergic fibers of pars reticulata. The α-synuclein increase may be a physiological temporal response to DA accumulation and/or to cell damage, but the simultaneous presence of α-synuclein and DA in the cell cytoplasm at concentration higher than normal is not exempt from risk. In fact, their incubation in a free cell system gives a stable dimerized form of α-synuclein that has been described as the critical rate-limiting step for its abnormal fibrillation.     

Recirculation of “B” Lymphocytes in Immunized Rats

THYMUS-derived, T lymphocytes and bursa-equivalent B lymphocytes cooperate in the initiation of the humoral antibody response of mammals to a variety of antigens^1–5. The B cells are antibody-forming-cell precursors (AFCP) and the T cells are helper cells which serve to augment the antibody response produced by the precursor cells⁶. Mitchison and his colleagues^4,5,7 have shown that the interaction between carrier and hapten-primed cells in the adoptive secondary antibody response to hapten-protein conjugates is an example of cooperation between T and B lymphocytes respectively.     

Chronic (3-Weeks) Treatment of Estrogen (17β-Estradiol) Enhances Working and Reference Memory in Ovariectomized Rats: Role of Acetylcholine

Recently there has been a growing interest in the effects of estrogen on cognitive functions. In this study, we aimed to examine 17β-estradiol treatment on working and reference memory in ovariectomized rats. We also examined the changes in the acetylcholine (ACh) levels in the brain areas associated with learning and memory. The study was performed on Sprague–Dawley type 3-month-old female rats. The rats were divided into four groups as control, ovariectomy (OVX), and OVX and estrogen treatment (10 µg/day i.p. 17β-estradiol) groups for 3 (OVX + E3) and 21 days OVX + E21). The rats were trained on eight arm radial maze task with eight arms baited to assess spatial memory, in addition four arms baited to assess both working and reference memory performances. The electron microscope images of the ACh vesicles in the frontal cortex, temporal cortex and hippocampus areas of the brain which are important regions for learning and memory were screened. Results showed that long term 17β-estradiol treatment has positive effects on both reference memory and working memory and that ACh vesicles increased in the examined brain areas, especially in hippocampus. Our results suggest that 3 weeks 17β-estradiol treatment may have an ameliorative effect on the memory through the central cholinergic system.

     

Trace Elements Status in Selenium-Deficient Rats—Interaction with Cadmium

Although the metabolic and toxicological interactions between essential element selenium (Se) and toxic element cadmium (Cd) have been reported for a long time, the experimental studies explored mostly acute, high-dose interactions. Limited data are available regarding the effects of Se-deficiency on toxicokinetics of cadmium, as well as on the levels of key trace elements—copper, zinc, and iron. In the present study, male and female Wistar weanling rats (n = 40/41) were fed either Se-deficient or Se-adequate diet (<0.06 or 0.14 mg Se per kilogram diet, respectively) for 12 weeks, and from week 9 were drinking water containing 0 or 50 mg Cd/l as cadmium chloride. At the end of the 12-week period, trace element concentrations were estimated by AAS. Selenium-deficient rats of both genders showed significantly lower accumulation of cadmium in the liver, compared to Se-adequate rats. Zinc and iron hepatic levels were not affected by Se-deficiency. However, a significant elevation of copper was found in the liver of Se-deficient rats of both genders. Cadmium supplementation increased zinc and decreased iron hepatic level, regardless of Se status and decreased copper concentration in Se-adequate rats. Se-deficiency was also found to influence the effectiveness of cadmium mobilization in male rats.     

Effects of Pyrazinamide on Tryptophan–Niacin Conversion in Rats

In the course of our screening for a new anti-tumor substance, the bisabolane sesquiterpenoid endoperoxide, 3,6-epidioxy-1,10-bisaboladiene (EDBD), was isolated from the edible wild-plant, Cacalia delphiniifolia. EDBD showed cytotoxicity toward human chronic myelogenous leukemia K562 and human prostate carcinoma LNCaP cell lines with IC₅₀ values of 9.1 μM and 23.4 μM, respectively. DNA fragmentation and condensation of chromatin, the hallmarks of apoptosis, appeared in K562 cells after an 18-h treatment with EDBD. α-Curcumene, a bisabolane sesquiterpene that lacks the endoperoxide moiety of EDBD, also showed cytotoxicity toward both K562 and LNCaP cell lines at over a 10-times higher dose than that of EDBD. The results indicate the importance of the endoperoxide structure within EDBD to its anti-tumor activity in vitro.     

Does Pilocarpine-Induced Epilepsy in Adult Rats Require Status epilepticus?

Pilocarpine-induced seizures in rats provide a widely animal model of temporal lobe epilepsy. Some evidences reported in the literature suggest that at least 1 h of status epilepticus (SE) is required to produce subsequent chronic phase, due to the SE-related acute neuronal damage. However, recent data seems to indicate that neuro-inflammation plays a crucial role in epileptogenesis, modulating secondarily a neuronal insult. For this reason, we decided to test the following hypotheses: a) whether pilocarpine-injected rats that did not develop SE can exhibit long-term chronic spontaneous recurrent seizures (SRS) and b) whether acute neurodegeneration is mandatory to obtain chronic epilepsy. Therefore, we compared animals injected with the same dose of pilocarpine that developed or did not SE, and saline treated rats. We used telemetric acquisition of EEG as long-term monitoring system to evaluate the occurrence of seizures in non-SE pilocarpineinjected animals. Furthermore, histology and MRI analysis were applied in order to detect neuronal injury and neuropathological signs. Our observations indicate that non-SE rats exhibit SRS almost 8 (+/22) months after pilocarpine-injection, independently to the absence of initial acute neuronal injury. This is the first time reported that pilocarpine injected rats without developing SE, can experience SRS after a long latency period resembling human pathology. Thus, we strongly emphasize the important meaning of including these animals to model human epileptogenesis in pilocarpine induced epilepsy.     

Improved Learning and Memory in Aged Mice Deficient in Amyloid β-Degrading Neutral Endopeptidase

Background

Neutral endopeptidase, also known as neprilysin and abbreviated NEP, is considered to be one of the key enzymes in initial human amyloid-β (Aβ) degradation. The aim of our study was to explore the impact of NEP deficiency on the initial development of dementia-like symptoms in mice.

Methodology/Principal Findings

We found that while endogenous Aβ concentrations were elevated in the brains of NEP-knockout mice at all investigated age groups, immunohistochemical analysis using monoclonal antibodies did not detect any Aβ deposits even in old NEP knockout mice. Surprisingly, tests of learning and memory revealed that the ability to learn was not reduced in old NEP-deficient mice but instead had significantly improved, and sustained learning and memory in the aged mice was congruent with improved long-term potentiation (LTP) in brain slices of the hippocampus and lateral amygdala. Our data suggests a beneficial effect of pharmacological inhibition of cerebral NEP on learning and memory in mice due to the accumulation of peptides other than Aβ degradable by NEP. By conducting degradation studies and peptide measurements in the brain of both genotypes, we identified two neuropeptide candidates, glucagon-like peptide 1 and galanin, as first potential candidates to be involved in the improved learning in aged NEP-deficient mice.

Conclusions/Significance

Thus, the existence of peptides targeted by NEP that improve learning and memory in older individuals may represent a promising avenue for the treatment of neurodegenerative diseases.     

Involvement of Spinal PKMζ Expression and Phosphorylation in Remifentanil-Induced Long-Term Hyperalgesia in Rats

Up-regulation of GluN2B-containing N-methyl-d-aspartate receptors (NMDARs) expression and trafficking is the key mechanism for remifentanil-induced hyperalgesia (RIH), nevertheless, the signaling pathway and pivotal proteins involved in RIH remain equivocal. PKMζ, an isoform of protein kinase C (PKC), maintains pain memory storage in neuropathic pain and inflammatory pain, which plays a parallel role regulated by NMDARs in long-term memory trace. In the present study, Zeta Inhibitory Peptide (ZIP), a PKMζ inhibitor, and a selective GluN2B antagonist Ro-256981 are injected intrathecally before remifentanil infusion (1 μg kg^?1 min^?1 for 1 h, iv) in order to detect whether GluN2B contributes to RIH through affecting synthesis and activity of PKMζ in spinal dorsal horn. Nociceptive tests are measured by Paw withdrawal mechanical threshold (PWT) and paw withdrawal thermal latency (PWL). The L₄–L₆ segments of dorsal horn taken from rats with RIH are for determining expression of PKMζ and pPKMζ by Western blot and immunohistochemistry. Our data suggest that remifentanil infusion causes an increase of PKMζ in expression and phosphorylation in rats with nociceptive sensitization, beginning at 2 h, peaked at 2 days, and returned to basal level at 7 days. ZIP (10 ng) could block behavioral sensitization induced by remifentanil. Ro25-6981 dosage-dependently attenuated mechanical and thermal hyperalgesia and reversed expression of PKMζ and pPKMζ, indicating that GluN2B-containing NMDA receptor facilitates development of RIH through mediating expression and activity of spinal PKMζ in rats. Although detailed mechanisms require further comprehensive study, the preventive role of Ro25-6981 and ZIP provide novel options for the effective precaution of RIH in clinics.     

GSK-3β Is Required for Memory Reconsolidation in Adult Brain

Activation of GSK-3β is presumed to be involved in various neurodegenerative diseases, including Alzheimer''s disease (AD), which is characterized by memory disturbances during early stages of the disease. The normal function of GSK-3β in adult brain is not well understood. Here, we analyzed the ability of heterozygote GSK-3β knockout (GSK+/−) mice to form memories. In the Morris water maze (MWM), learning and memory performance of GSK+/− mice was no different from that of wild-type (WT) mice for the first 3 days of training. With continued learning on subsequent days, however, retrograde amnesia was induced in GSK+/− mice, suggesting that GSK+/− mice might be impaired in their ability to form long-term memories. In contextual fear conditioning (CFC), context memory was normally consolidated in GSK+/− mice, but once the original memory was reactivated, they showed reduced freezing, suggesting that GSK+/− mice had impaired memory reconsolidation. Biochemical analysis showed that GSK-3β was activated after memory reactivation in WT mice. Intraperitoneal injection of a GSK-3 inhibitor before memory reactivation impaired memory reconsolidation in WT mice. These results suggest that memory reconsolidation requires activation of GSK-3β in the adult brain.     

“Chromosome Memory” of Parental Genomes in Embryonic Hybrid Cells

In the hybrid cells obtained by fusion of embryonic stem cells with adult differentiated cells, homologous chromosomes are in two ontogenetic configurations: pluripotent and differentiated. In order to assess the role of cis- and trans-regulation in the maintenance of these states, we studied a set of clones of hybrid cells of the type embryonic stem cells–splenocytes and used two approaches: segregation of parental chromosomes and comparison of pluripotency of the past hybrid cells and embryonic stem cells. The segregation test showed that the hybrid cells lost only the homologs of the somatic partner and this process was sharply accelerated when the cells were cultivated in nonselective conditions, thus suggesting the full or partial preservation of the initial differences in the organization of parental homologs. The descendants of the former hybrid cells, which had the karyotype similar to that of embryonic stem cells, demonstrated the level of pluripotency, comparable with that of embryonic stem cells despite the long-term effect of trans-acting factors from the somatic partner in the genome of hybrid cells. The data obtained are interpreted in the framework of the concept of chromosome memory, in the maintenance of which the key role is played bycis-regulatory factors.     

How Bad Is Aluminum Exposure to Reproductive Parameters in Rats?

The objective of this study was to evaluate influence of dietary palygorskite (Pal) supplementation on growth performance, mineral accumulations in the tissues (livers, kidneys, and muscles), antioxidant capacities, and meat quality of broilers fed lead (Pb)-contaminated diet. One-hundred forty-four male broiler chicks were randomly divided into three treatment groups, receiving a corn-soybean meal basal diet (the control group), the basal diet contaminated with 10 mg/kg Pb (the Pb group), and the basal diet with 10-g/kg Pal supplementation and 10-mg/kg Pb contamination (the Pal/Pb group) from 1 to 42 days of age, respectively. Treatments did not affect growth performance of broilers in the 42-day study (P > 0.05). Compared with the control group, Pb contamination increased Pb accumulation in the livers, kidneys, and muscles (P < 0.05); elevated malondialdehyde accumulation in the livers, kidneys, and breast muscles; glutathione peroxidase activity in the livers and superoxide dismutase activity in the kidneys (P < 0.05); exacerbated drip loss in the pectoralis muscles (P < 0.05); and reduced glutathione peroxidase activity in the pectoralis muscles (P < 0.05) of broilers at 42 days of age. The values of these parameters were reversed in the Pal/Pb group to levels comparable with those in the control group (P < 0.05). Additionally, Pal supplementation reduced redness value in the pectoralis muscles (P < 0.05), and decreased Cu concentration in the pectoralis muscles and livers at 42 days of age as well as its accumulation in the kidneys at both 21 and 42 days of age compared with the other two groups (P < 0.05). The results suggested that dietary Pal supplementation would decrease Pb residue in the tissues, alleviate oxidative stress, and affect meat quality of broilers exposed to Pb.