Dose Of LiCl Research Articles

New insight into the toxic effects of lithium in the ragworm Perinereis cultrifera as revealed by lipidomic biomarkers, redox status, and histopathological features.

Lithium (Li) is a toxic monovalent alkaline metal used in household items common to industrial applications. The present work was aimed at investigating the potential toxic effects of LiCl on the redox status, fatty acid composition, and histological aspects of the marine ragworm Perinereis cultrifera. Sea worms were exposed to LiCl graded doses (20, 40, and 80mg/L) for 48h. Compared with the control group, the saturated fatty acids (SFA) decreased while monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA) increased upon exposure to LiCl. The increase in PUFA n-3 and PUFA n-6 was concomitant to an increase in docosahexaenoic (DHA: C22:6n-3), eicosapentaenoic (EPA: C20:5n-3), and docosapentaenoic acid (C22:5n-6) fatty acids. Results showed that LiCl-treated specimens accumulate lithium with increasing exposure gradient. Indeed, the exposure to LiCl doses promoted oxidative stress with an increase of the ferric reducing antioxidant power (FRAP), malondialdehyde (MDA), hydrogen peroxide (H2O2), advanced oxidation protein product (AOPP), and protein carbonyl (PCO) as well as the enzymatic and non-enzymatic antioxidants (non-protein thiols (NPSH), catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione S-transferase (GST), and metallothionein (MT)) levels in all treated groups. Our biochemical findings have been affirmed by the histopathological observations showing hyperplasia and loss of the intestine structure in treated specimens. Overall, our findings give new insights on the toxic effect of LiCl on the redox status of P. cultrifera body tissue and highlighted the usefulness of the FA composition as an early sensitive bioindicators to better understand LiCl mechanism of toxicity in marine polychaetes.

Voluntary wheel running and lithium supplementation improves muscle force production without altering SERCA function in the D2 mdx mouse

Background: Duchenne muscular dystrophy (DMD) is an X-linked recessive muscle wasting disease that, with no cure, leads to early mortality (30-40 years of age). Previous studies have shown that voluntary wheel running can mitigate disease pathology and improve muscle contractility in the preclinical mdx mouse model. We have shown that treating wild-type (WT) mice with a low dose of lithium, a natural inhibitor of glycogen synthase kinase 3, enhanced soleus (SOL) and extensor digitorum longus (EDL) muscle contractility. Our objectives were to test our hypothesis of whether combining VWR with low dose lithium supplementation could provide further benefits to muscle contractility in DBA/2J mdx mice – a model that displays far worse disease severity compared with the traditional C57BL/10 mdx mouse. We also examined if this combined intervention could improve muscle calcium regulation in muscle by assessing sarco(endo)plasmic reticulum Ca 2+ -ATPase (SERCA) function as we have recently shown that SERCA function is drastically impaired in D2 mdx mice, which could also contribute to deficits in muscle force production. Methods: Young (5-6 week old) male D2 WT and mdx mice were ordered from Jackson Laboratory. D2 mdx mice were assigned into each of the following conditions: sedentary ( mdxSED), voluntary wheel running ( mdxVWR), or VWR+lithium chloride ( mdxVWR+Li). The VWR groups were given access to a cagewheel for a total of 6 weeks, and the mdxVWR+Li group were provided a low dose of LiCl (50 mg/kg/day) via drinking water for the entire duration of the study. After 6 weeks, all mice were euthanized and the SOL and EDL muscles were subjected to force-frequency curve analysis (1-160 Hz). SERCA function was assessed by performing SERCA activity assays and a Ca 2+ uptake assay. Results: In the SOL, the combined effect of lithium and VWR improved specific force production to near WT levels across submaximal and maximal frequencies. Specific force production in the mdxVWR+Li group was significantly greater compared to the mdxSED group at all stimulation frequencies. Rate of SOL force development was significantly lower in all mdx groups compared to WT. However, rate of relaxation was only significantly reduced (vs. WT) within the mdxSED group, suggesting that VWR and VWR+Li improved relaxation. This could not be explained by any changes in SERCA function. In the EDL, specific force was significantly (p<0.001) reduced across mdx groups compared to WT; however, of all mdx groups, the mdxVWR+Li produced the most force and was significantly different compared to mdxVWR at 80 and 160 Hz. Rates of force development and relaxation were all slower in all mdx groups compared to WT, and there were no differences in SERCA activity. Conclusions: Combining lithium supplementation with VWR improved SOL and EDL force production in the D2 mdx mouse without any changes to SERCA function. V.A.F is a Tier 2 Canada Research Chair in Muscle Plasticity and Tissue-Remodelling. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Evaluation of Sex Differences in the Potential of Δ9-Tetrahydrocannabinol, Cannabidiol, Cannabidiolic Acid, and Oleoyl Alanine to Reduce Nausea-Induced Conditioned Gaping Reactions in Sprague-Dawley Rats.

Introduction: Cancer patients report nausea as a side effect of their chemotherapy treatment. Using the pre-clinical rodent model of acute nausea-lithium chloride (LiCl)-induced conditioned gaping-our group has demonstrated that exogenous cannabinoids may have antinausea potential. Materials and Methods: With the goal of evaluating the role of sex as a factor in pre-clinical research, we first compared the conditioned gaping reactions produced by varying doses of LiCl in male and female rats using the taste reactivity test (Experiment 1). Results: LiCl produced dose-dependent conditioned gaping similarly in male and female rats with the highest dose (127.2 mg/kg) producing robust conditioned gaping, with this dose used in subsequent experiments. Next, we examined the antinausea potential of THC (Experiment 2), CBD (Experiment 3), cannabidiolic acid (CBDA; Experiment 4) and oleoyl alanine (OlAla; Experiment 5) in both male and female rats. THC, CBD, CBDA, and OlAla dose dependently reduced conditioned gaping in both male and female rats in a similar manner. Conclusions: These results suggest that cannabinoids may be equally effective in treating nausea in both males and females.

Lithium Inhibition of GSK3 Uncouples SERCA Transport Efficiency in C2C12 Cells and Alters Energy Expenditure in vivo

Adaptive thermogenesis is a cellular process that accelerates energy expenditure while increasing heat production in response to prolonged cold exposure or caloric excess. Enhancing adaptive thermogenesis mechanisms may assist in combatting diet-induced obesity. Skeletal muscle via sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) uncoupling and brown/beige adipose via mitochondrial uncoupling are the two primary sites for adaptive thermogenesis in mammals. Recent evidence has shown that glycogen synthase kinase 3 (GSK3) negatively regulates adipose-based thermogenesis by repressing uncoupling protein-1 expression in brown adipocytes. However, to our knowledge, there have been no studies examining whether GSK3 also negatively regulates muscle-based thermogenesis via SERCA uncoupling. Lithium (LiCl), a known GSK3 inhibitor, has previously been shown to offset diet induced obesity in rodents. Thus, we examined the effect of LiCl treatment on energy expenditure first utilizing a cellular model followed by an in vivo mouse model. To this end, C2C12 cells were treated with 0.5 mM LiCl at the onset of differentiation for 7 days. SERCA coupling ratios were obtained by measuring the rates of Ca2+ uptake using a fluorophore-based assay and the rates of ATP hydrolysis using a spectrophotometric assay at physiological Ca2+ concentrations. Our results show that when treated with LiCl, C2C12 cells had significantly reduced Ca2+ uptake (-59%, p = 0.03) with no change in SERCA activity. This led to a significant reduction in SERCA's coupling ratio (p = 0.04). These results are consistent with our previous findings suggesting that LiCl increases basal respiration and SERCA's energy contribution in C2C12 cells. To support this increase in energy expenditure, our Western blot analyses identified that LiCl treatment inhibited GSK3 and showed increased levels of PGC-1a, the master regulator of mitochondrial biogenesis. To investigate the effects of LiCl on energy expenditure in vivo, adult male C57BL/6J mice were provided 10mg/kg/day LiCl via their drinking water for 6 weeks. We have previously shown that this dose and duration of LiCl significantly inhibits GSK3 in murine skeletal muscle. On the 6thweek of treatment, mice were housed in a Promethion metabolic cage system for a period of 48h to examine daily energy expenditure via indirect calorimetry. Metabolic cage data revealed that LiCl fed mice had significantly increased energy expenditure (kcal/kg/hr; p = 0.008) compared to controls; however, the contribution from muscle-based thermogenesis and SERCA uncoupling still needs to be examined. Altogether, these data suggest that GSK3 inhibition via LiCl treatment augments SERCA energy expenditure in muscle by uncoupling Ca2+ transport from ATP hydrolysis. Future studies will determine whether our in vitro findings translate to the in vivo setting.

High dose lithium chloride causes colitis through activating F4/80 positive macrophages and inhibiting expression of Pigr and Claudin-15 in the colon of mice

ObjectiveLithium chloride (LiCl) was a mood stabilizer for bipolar affective disorders and it could activate Wnt/β-catenin signaling pathway both in vivo and in vitro. Colon is one of a very susceptible tissues to Wnt signaling pathway, and so it would be very essential to explore the toxic effect of a high dose of LiCl on colon. MethodsC57BL/6 mice were injected intraperitoneally with 200 mg/kg LiCl one dose a day for 5 days to activate Wnt signal pathway in intestines. H&E staining was used to assess the colonic tissues of mice treated with high dose of LiCl. The expression of inflammation-associated genes and tight junction-associated genes in colons was measured using qPCR, Western blot and immunostaining methods. The gut microbiome was tested through 16S rDNA gene analysis. ResultsThe differentiation of enteroendocrine cells in colon was inhibited by treatment of 200 mg/kg LiCl. The F4/80 positive macrophages in colon were activated by high dose of LiCl, and migrated from the submucosa to the lamina propria. The expression of pro-inflammatory genes TNFα and IL-1β was increased in the colon of high dose of LiCl treated mice. Clostridium_sp_k4410MGS_306 and Prevotellaceae_UCG_001 were specific and predominant for the high dose of LiCl treated mice. The expression of IgA coding genes, Pigr and Claudin-15 was significantly decreased in the colon tissues of the high dose of LiCl treated mice. Conclusion200 mg/kg LiCl might cause the inflammation in colon of mice through activating F4/80 positive macrophages and inhibiting the expression of IgA coding genes in plasma cells and the expression of Pigr and Claudin-15 in colonic epithelial cells, providing evidences for the toxic effects of high dose of LiCl on colon.

Short-term memory reactivation of a weak CS-US association promotes long-term memory persistence in conditioned odor aversion.

In conditioned odor aversion (COA), the association of a tasteless odorized solution (the conditioned stimulus [CS]) with an intraperitoneal injection of LiCl (the unconditioned stimulus [US[), which produces visceral malaise, results in its future avoidance. The strength of this associative memory is mainly dependent on two parameters, that is, the strength of the US and the interstimuli interval (ISI). In rats, COA has been observed only with ISIs of ≤15 min and LiCl (0.15 M) doses of 2.0% of bodyweight, when tested 48 h after acquisition (long-term memory [LTM]). However, we previously reported a robust aversion in rats trained with ISIs up to 60 min when tested 4 h after acquisition (short-term memory [STM]). Since memories get reactivated during retrieval, in the current study we hypothesized that testing for STM would reactivate this COA trace, strengthening its LTM. For this, we compared the LTM of rats trained with long ISIs or low doses of LiCl initially tested for STM with that of rats tested for LTM only. Interestingly, rats conditioned under parameters sufficient to produce STM, but not LTM, showed a reliable LTM when first tested for STM. These observations suggest that under suboptimal training conditions, such as long ISIs or low US intensities, a CS–US association is established but requires reactivation in the short-term in order to persist in the long-term.

Chitin Hydrogels Prepared at Various Lithium Chloride/N,N-Dimethylacetamide Solutions by Water Vapor-Induced Phase Inversion

Chitin was chemically extracted from crab shells and then dissolved in N,N-dimethylacetamide (DMAc) solvent with lithium chloride (LiCl) at 3, 5, 7, and 10%. The concentrated chitin-DMAc/LiCl solutions were used for the preparation of chitin hydrogels by water vapor-induced phase inversion at 20°C. The coagulation process was investigated while altering the concentration of LiCl in the DMAc solution. The shear viscosity of the chitin solution increased with higher LiCl amounts and decreased when the concentration of LiCl was reduced by adding water to the chitin solution, implying high LiCl concentration delayed the coagulation of chitin solution in the presence of water. The viscoelasticity of the chitin solutions indicated the gel formation intensification was dependent on the dose of LiCl and chitin in the DMAc solution. After the chitin solution was coagulated, the resultant hydrogels had water contents of 387–461% and the tensile strength varied from 285 to 400 kPa when the concentration of LiCl in the hydrogel was adjusted to 3% and 7%, respectively. As for viscoelasticity, the complex modulus of the chitin hydrogels indicated that the increment of the LiCl concentration up to 7% formed the tight hydrogels. Atomic force microscopic (AFM) image revealed the formation of the entanglement network and larger domains of the aggregated chitin segments. However, the hydrogel prepared at 10% LiCl in DMAc solution exhibited weak mechanical properties due to the loose hydrogel networking caused by the strong aggregation of the chitin segments.

The role of WNT/β-catenin signaling pathway and glutamine metabolism in the pathogenesis of CCl4-induced liver fibrosis: Repositioning of niclosamide and concerns about lithium

BackgroundLiver fibrosis is a serious health problem which may lead to advanced liver cirrhosis and hepatocellular carcinoma. ObjectiveThe present study aimed to investigate the role of Wnt/β-catenin signaling pathway and glutamine aminohydrolase enzyme (l-glutaminase) in the pathogenesis of liver fibrosis and the potential benefits of niclosamide in treating liver fibrosis. MethodsNinety male Albino rats were divided into 6 equal groups (n = 15) as follows: a normal control group (NC), CCl4-only treated group (Fib.) which received 1 mg/kg CCl4 two times weekly, niclosamide-treated group (Niclo.) which received 5 mg/kg of niclosamide one time daily, lithium chloride-treated group (LiCl) which received 100 mg/kg of LiCl one time daily, niclosamide-and-CCl4-treated group (Niclo. + Fib.) which received same doses of niclosamide and CCl4 given to other groups, and finally lithium chloride-and-CCl4-treated rat group (LiCl + Fib.) which received same doses of LiCl and CCl4 given to other groups. All treatments were administered orally for 8 weeks. Liver tissue was assessed for l-hydroxyproline, beta-catenin (β-catenin), l-glutaminase activity, as well as the gene expression of transforming growth factor beta-1 (TGF-β1) and Dishevelled-2 (Dvl2). Histopathological and immunohistochemical analyses of alpha smooth muscle actin α-SMA were performed. Serum alanine transaminase (ALT), aspartate transaminase (AST), and total bilirubin were measured. ResultsThe group of niclosamide-and-CCl4-treated rats showed a significant decrease in total bilirubin, ALT and AST, β-catenin, l-hydroxyproline, l-glutaminase activity, and gene expression of TGF-β1 and Dvl2. Moreover, the liver tissue in this group of rats showed mild α-SMA reactivity compared with the rats treated with CCl4 only (fibrosis group). On the other hand, lithium chloride-and-CCl4-treated rats showed a significant increase in liver indices, TGF-β1 expression, β-catenin, l-hydroxyproline, and l-glutaminase activity with severe α-SMA reactivity and apoptosis in the liver tissue. ConclusionsNiclosamide protected rats against liver fibrosis by inhibiting the Wnt/β-catenin pathway and glutaminolysis.

Differential gene expression by lithium chloride induction of adipose-derived stem cells into neural phenotype cells.

Objective(s):Adipose-derived stem cells (ADSCs), with suitable and easy access, are multipotential cells that have the ability for differentiation into other mesodermal and transdifferentiate into neural phenotype cells. In this study, Lithium chloride (LiCl) was used for in vitro transdifferentiation of rat ADSCs into neuron-like cells (NLCs).Materials and Methods:ADSCs were isolated from the rats’ perinephric region using Dulbecco΄s Modified Eagle΄s Medium (DMEM) with Fetal Bovine Serum (FBS), cultured for 3 passages, characterized by flowcytometry and differentiation into adipogenic and osteogenic phenotypes. The ADSCs were exposed to 0.1, 0.5, 1, 1.5, 2, 5, and 10 millimolar (mM) LiCl without serum for 24 hr. The optimum dose of LiCl was selected according the maximum viability of cells. The expression of neurofilament light chain (NfL), neurofilament high chain (NfH), and nestin was evaluated by immunocytochemistry. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to evaluate the amount of synaptophysin, neurogenin-1, neuroD1, NfL, NfH, and nestin genes’ expression in ADSCs and NLCs.Results:The optimum dose of LiCl was 1 mM in 24 hr. The transdifferentiated ADSCs showed cytoplasmic extension with synapse-like formation. Synaptophysin, neurogenin-1, neuroD1, NfL, NfH, and nestin genes were significantly expressed more in NLCs than in ADSCs.Conclusion:LiCl can induce ADSCs into neural phenotype cells with higher expression of neural and neuronal genes.

A Low-Therapeutic Dose of Lithium Inhibits GSK3 and Enhances Myoblast Fusion in C2C12 Cells.

Glycogen synthase kinase 3 (GSK3) slows myogenic differentiation and myoblast fusion partly by inhibiting the Wnt/β-catenin signaling pathway. Lithium, a common medication for bipolar disorder, inhibits GSK3 via Mg+ competition and increased Ser21 (GSK3α) or Ser9 (GSK3β) phosphorylation, leading to enhanced myoblast fusion and myogenic differentiation. However, previous studies demonstrating the effect of lithium on GSK3 have used concentrations up to 10 mM, which greatly exceeds concentrations measured in the serum of patients being treated for bipolar disorder (0.5–1.2 mM). Here, we determined whether a low-therapeutic (0.5 mM) dose of lithium could promote myoblast fusion and myogenic differentiation in C2C12 cells. C2C12 myotubes differentiated for three days in media containing 0.5 mM lithium chloride (LiCl) had significantly higher GSK3β (ser9) and GSK3α (ser21) phosphorylation compared with control myotubes differentiated in the same media without LiCl (+2–2.5 fold, p < 0.05), a result associated with an increase in total β-catenin. To further demonstrate that 0.5 mM LiCl inhibited GSK3 activity, we also developed a novel GSK3-specific activity assay. Using this enzyme-linked spectrophotometric assay, we showed that 0.5 mM LiCl-treated myotubes had significantly reduced GSK3 activity (−86%, p < 0.001). Correspondingly, 0.5 mM LiCl treated myotubes had a higher myoblast fusion index compared with control (p < 0.001) and significantly higher levels of markers of myogenesis (myogenin, +3-fold, p < 0.001) and myogenic differentiation (myosin heavy chain, +10-fold, p < 0.001). These results indicate that a low-therapeutic dose of LiCl is sufficient to promote myoblast fusion and myogenic differentiation in muscle cells, which has implications for the treatment of several myopathic conditions.

Nitric oxide and glutamate are contributors of anti-seizure activity of rubidium chloride: A comparison with lithium

The neuro-protective effects of rubidium and lithium as alkali metals have been reported for different central nervous system dysfunctions including mania and depression. The aim of this study was evaluating as well as comparing the effects of rubidium chloride (RbCl) and lithium chloride (LiCl) on different seizures paradigms in mice and determining the involvement of NMDA receptors and nitrergic pathway.To assess the seizures threshold, animals received intravenous pentylenetetrazole (PTZ, 0.5%; 1 mL/min). Male NMRI mice (6–8 weeks) received intraperitoneal (i.p.) injections of different doses of RbCl and LiCl. Doses greater than 10 mg/kg of RbCl showed a significant anticonvulsant activity 60 min after administration; the anticonvulsant effects of LiCl was observed at the doses more than 5 mg/kg and after 30 min in PTZ-induced seizure threshold. But, RbCl (10, 20 mg/kg, i.p) or LiCl (5, 10 mg/kg, i.p) injection did not induce protection against maximal electroshock (MES) or intraperitoneal injection of PTZ lethal dose (80 mg/kg)-induced seizure models.Pre-treatment with L-NAME (non-selective nitric oxide synthase (NOS) inhibitor, 10 mg/kg; i.p.) and 7-nitroindazole (selective neuronal NOS inhibitor, 30 mg/kg; i.p.) enhanced the anticonvulsive effects of both RbCl (5 mg/kg, i.p.) and LiCl (1 mg/kg, i.p.) in PTZ-induced seizure threshold model. Injection of MK-801 (NMDA receptor antagonist, 0.05 mg/kg; i.p.) before RbCl (5 mg/kg, i.p.; P < 0.001) and LiCl (1 mg/kg, i.p.; P < 0.001) administration increased the anti-seizure activity. But, treatment with L-arginine (precursor of nitric oxide, 100 mg/kg; i.p.) decreased the seizure threshold of both RbCl (20 mg/kg, i.p.; P < 0.001) and LiCl (10 mg/kg, i.p.; P < 0.001). Measurement of nitrite levels in hippocampus of animals revealed a remarkable reduction after treatment with RbCl (20 mg/kg, i.p; P < 0.05) and LiCl (10 mg/kg, i.p; P < 0.01).To conclude, rubidium may protect central nervous system against seizures in PTZ-induced seizures threshold model through NMDA/nitrergic pathways with a similarity to lithium effects in mice.

Conditioned taste aversion versus avoidance: A re-examination of the separate processes hypothesis.

Rats not only avoid ingesting a substance associated with LiCl toxicosis, but they display rejection reflexes (e.g., gapes) to its taste; this latter response is thought to reflect disgust or taste aversion. Prior work has shown that rats also avoid consuming foods/fluids associated with other adverse gastrointestinal (GI) effects like lactose indigestion but without the concomitant change in oromotor responses (taste reactivity; TR) indicative of aversion. Because of interpretive limitations of the methods used in those studies, we revisited the taste aversion-avoidance distinction with a design that minimized non-treatment differences among groups. Effects on intake and preference (Experiments 1a, 1b, and 2), as well as consummatory (TR, Experiment 1a and 1b) and appetitive (Progressive Ratio, Experiment 2) behaviors to the taste stimulus were assessed after training. In both experiments, rats were trained to associate 0.2% saccharin (CS) with intraduodenal infusions of LiCl, Lactose, or NaCl control. Rats trained with 18% lactose, 0.3 and 1.5 mEq/kg dose of LiCl subsequently avoided the taste CS in post-training single-bottle intake tests and two-bottle choice tests. However, only those trained with 1.5 mEq/kg LiCl displayed post-conditioning increases in taste CS-elicited aversive TR (Experiment 1a and 1b). This dose of LiCl also led to reductions in breakpoint for saccharin. The fact that conditioned avoidance is not always accompanied by changes in other common appetitive and/or consummatory indices of ingestive motivation further supports a functional dissociation between these processes, and highlights the intricacies of visceral influences on taste-guided ingestive motivation.

The Wnt/β-Catenin/LEF1 Pathway Promotes Cell Proliferation at Least in Part Through Direct Upregulation of miR-17-92 Cluster

The miR-17-92 cluster is involved in animal development and homeostasis, and its dysregulation leads to human diseases such as cancer. In the present study, we investigated the functional link between miR-17-92 cluster and Wnt/β-catenin signaling pathway in ICP2 and DF1 cells. We demonstrated that ectopic expression of either LEF1 or β-catenin increased the promoter activity of the miR-17-92 cluster host gene (MIR17HG) and combined ectopic expression of LEF1 and β-catenin further enhanced the promoter activity; while knockdown of either LEF1 or β-catenin reduced the MIR17HG promoter activity. Both LEF1 and β-catenin could directly bind to the MIR17HG promoter. Furthermore, we demonstrated that low doses of lithium chloride (LiCl), an activator of Wnt/β-catenin signaling pathway, increased MIR17HG promoter activity and the endogenous expression of the miR-17-92 cluster, while high doses of LiCl had the opposite effects. Treatment with XAV-939, an inactivator of the Wnt/β-catenin pathway, reduced the endogenous expression of miR-17-92 cluster. Finally, we found that low doses of LiCl promoted the proliferation of ICP2 and DF1 cells, while high doses of LiCl inhibited the proliferation of ICP2 and DF1 cells. Taken together, our results reveal that MIR17HG is a target of LEF1 and the Wnt/β-catenin pathway and suggest that the miR-17-92 cluster may, at least in part, mediate the proliferation-promoting effect of the Wnt/β-catenin pathway in cell proliferation.

Ameliorative effect of lithium chloride against d-galactose induced behavioral and memory impairment, oxidative stress and alteration in serotonin function in rats.

Aging is a phenomenon that all living organisms surely face. d-galactose (D-gal) has been used to develop an aging model of brain. Lithium (Li) has been proposed to have neuroprotective properties in relation to several neurological disorders. The goal of the current studyis to evaluate the effect of Lithium Chloride (LiCl) on D-gal induced neurological disorders and oxidative stress. Rats were treated with D-gal at a dose of 300 mg/ml/kg and various doses of LiCl (20, 40 and 80 mg/ml/kg) for 14 days. After that behavioral analysis (Elevated plus maze (EPM); Light dark box test (LDT); Morris water maze (MWM); Forced swim test (FST)) were performed. Animals were decapitated after behavioral tests and brain samples were collected for biochemical (malondialdehyde (MDA); superoxide dismutase (SOD); catalase (CAT); glutathione peroxidase (GPx); acetylcholiesterase (AChE)) and neurochemical analysis (5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA)). The results showed that administration of LiCl at all doses ameliorates D-gal induced, decreased time spent in the open arm and light box in EPM and LDT respectively, increased immobility in FST, increased latency escape in MWM, increased MDA levels, decreased antioxidant enzyme, increased AChE activity and decreased 5-HT metabolism. In conclusion, the present study indicated that D-gal induced anxiety/depression like symptoms and memory impairment were ameliorated by LiCl (at all doses) possibly via its antioxidant effects and normalizing 5-HT function.

Conditioned aversive responses produced by delayed, but not immediate, exposure to cocaine and morphine in male Sprague-Dawley rats.

To determine the conditions under which tastes paired with delayed access to experimenter-delivered cocaine and morphine elicit a conditionally aversive affective state. The potential of saccharin paired with immediate access to cocaine (5, 10, 20mg/kg, sc and ip) and delayed (30 and 10min) access to cocaine (20mg/kg, scand ip) and morphine (10mg/kg, sc) to elicit a pattern of aversive responding in the taste reactivity test (Grill and Norgren 1978a) was evaluated. Cocaine-induced aversions were compared with those produced by a moderate dose of LiCl (50mg/kg). Finally, as an independent measure of cocaine withdrawal, the potential of exposure to saccharin paired with delayed access to cocaine to produce anxiogenic-like responding in the Light-Dark Emersion test was evaluated. Immediate access to cocaine did not produce conditioned aversion at any dose. Delayed (30 or 10min) access to sc cocaine (20mg/kg) produced robust conditioned aversion and delayed access to ip cocaine (20mg/kg; 30min) and to sc morphine (10mg/kg; 10min) produced weaker conditioned aversion. Yawning emerged as a potential withdrawal response in rats conditioned with delayed (30 min) access to 20mg/kg, sc, cocaine. Contextual cues did not produce conditioned aversion when paired with delayed access to sc cocaine (20mg/kg). Finally, exposure to saccharin paired with delayed access to cocaine produced anxiogenic-like responding in the Light-Dark Emersion test. Our results support the contention that a conditioned aversive state develops when a taste cue comes to predict the delayed availability of drugs of abuse.

Vitamin C as an antioxidant stabilizes the depressive brain

Lithium chloride treatment of neural diseases, such as manic-depressive disorder, causes impairment of anti-oxidative defense. It induces opposite effects on nitric oxide-related systems. Antioxidant supplementation (Ascorbic acid) with Li Cl could have a therapeutic effect in unstable patients. The present study investigates the modulation of NO-producing cells in the rat brain, due to Li Cl administration to experimental animals. We report here the potential that Vitamin C administration along with Li Cl has in counteracting anti-oxidative stress. In control animals and in animals injected with low dose Li Cl together with Vitamin C, there were large numbers of NO-producing neurons. This number significantly decreased in animals treated only with the low dose of Li Cl and those injected with the high dose of Li Cl together with vitamin C. Moreover, the smallest number of NO-producing cells appeared in animals treated with high dose Li Cl only. Therefore, we suggest that Vitamin C reduces Li Cl noxious effects (reduction of NO levels) and recommend administering it together with mood stabilizing drugs.

Unconditioned Stimulus Pre-exposure Interferes with Conditioned Taste Aversion in Rats

Pairing of a gustatory CS and the visceral US induced by lithium chloride (LiCl) results in reduction of consumption of CS solution. This reduction is termed conditioned taste aversion (CTA). Pre-exposure to CS alone prior to the conditioning disrupts the conditioning (latent inhibition) and so is pre-exposure to US. The US pre-exposure effect is hypothesized to be due to tolerance, habituation, or the associative blocking effect. We studied the dose effect of US pre-exposure on CTA. In 3 experiments, rats were exposed to 10 doses of LiCl before the CTA procedure that consisted of ingesting a saccharin solution (0.1%) followed by injection of a constant dose of LiCl (7.632mg/rat). If the pre-exposure reduced the associability of the US, the CTA should be less. Exp. 1: Pre-exposure to 4 or 8 ml/kg of 0.15M LiCl, 1, 2 or 4 days prior to CTA did not have effect. Exp. 2: Pre-exposure to 0.5, 1, 2, 16, 32 ml/kg 1-day prior disrupted CTA. Exp. 3: Pre-exposure to 1, 16, 20, 32, 40 ml/kg 2-days prior disrupted CTA. Summary: (a) The pre-exposure effect was the same whether the pre-exposure occurred 1 or 2 days prior to conditioning. (b) A clear dose-effect relationship existed. (c) The relation was not monotonic. (d) The lowest preexposure dose, 0.5ml/kg of 0.15M LiCl, not strong enough to induce CTA by itself, was effective. (e) Upon repeated pairing of CS and US, CTA became more pronounced. The results do not support the hypotheses mentioned earlier.

Using long-term averted goats for selective grazing in olive groves

Conditioned taste aversion (CTA) is a useful tool to modify animal feed preferences, allowing the implementation of selective grazing to control weeds in tree orchards without damaging the trees or affecting fruit production. LiCl is commonly used for inducing CTA. However, studies investigating the long-term persistence of CTA by LiCl in small ruminants are scarce. With this aim, we evaluated the efficiency of two LiCl doses (AV1 and AV2, 175 and 200 mg/kg BW, respectively) and a control (C, 0 mg/kg BW) for averting non-lactating dairy goats (n=15) to olive tree leaves. Aversion induction was reinforced on day 9 in those goats that consumed >10 g of olive leaves. Mid-term aversion effectiveness was assessed by five double-choice feeding tests (days 16, 24, 31, 38 and 53) of 30 min each, where 100 g of olive leaves were offered side-by-side with 390 g of Italian rye-grass (as-fed). Long-term aversion effectiveness was assessed in C, AV1 and AV2 goats by grazing for 30 min in paddocks with a simulated olive tree (days 59, 90, 121, 182 and 420). Moreover, C and AV2 goats were compared under on-field conditions (days 143, 211 and 363) in a commercial olive grove also for 30 min. The CTA proved to be established with a single LiCl dose in all goats and persisted for 4 and 55 days in AV1 and AV2 goats, respectively (P<0.001). However, 80% AV1 and 20% AV2 goats needed to be reinforced at day 9. When grazing under simulated olive tree and commercial olive grove conditions, the CTA goats, especially AV2 group, avoided the contact with the olive trees and minimally used a bipedal stance to feed leaves, than control goats. On average, time proportion spent consuming olive leaves and sprouts was much greater (P<0.05) for C (50.7±9.1%) than for AV1 (14.4±3.9%) and AV2 (3.1±0.9%). In conclusion, the 200 mg LiCl/kg BW dose was more effective than the 175 mg LiCl/kg BW dose for inducing an effective long-term CTA to olive tree leaves in goats.

Lithium chloride could aggravate brain injury caused by 3-nitropropionic acid.

Lithium, a well-known drug for the treatment of bipolar disorder, may also have the ability to reduce neurodegeneration and stimulate cell proliferation. Systemic injection of mitochondrial toxin 3-nitropropionic acid (3NPA) is known to induce a relatively selective, Huntington disease-like brain injury. The aim of this study was to determine the effect of lithium chloride (LiCl) on brain injury caused by 3NPA. Female adult Wistar rats were pre-treated with LiCl (127 mg/kg) 1 day before the first injection of 3NPA (28 mg/kg), and then for 8 days with the same treatment but receiving LiCl 1 hour before 3NPA. Control groups were pre-treated accordingly, with LiCl or with normal saline, but were not treated with 3NPA. Staining for cytochrome c oxidase activity and in situ hybridization autoradiography of synaptotagmin-4 and -7 mRNAs were used to evaluate brain injury caused by 3NPA. There was a significant reduction of body weight in the 3NPA+LiCl group (79%) compared to the 3NPA group (90%, p = 0.031) and both control groups (100%, p = 0.000). Densitometric evaluation of cytochrome c oxidase staining and in situ hybridization autoradiograms revealed that the pre-treatment with LiCl caused an increase in striatal lesion for about 40% (p = 0.049). Moreover, the lesion was observed also in the hippocampus of three animals from the 3NPA+LiCl group and in two animals from the 3NPA group. However, there were no differences between the LiCl and saline group in any of the measured parameters. We concluded that the pre-treatment with a relatively nontoxic dose of LiCl could aggravate brain injury caused by 3NPA.

The role of dose and restriction state on morphine-, cocaine-, and LiCl-induced suppression of saccharin intake: A comprehensive analysis

Rats avoid intake of a taste cue when paired with a drug of abuse or with the illness-inducing agent, lithium chloride (LiCl). Although progress has been made, it is difficult to compare the suppressive effects of abused agents and LiCl on intake of a gustatory conditioned stimulus (CS) because of the cross-laboratory use of different CSs, different unconditioned stimuli (USs), and different doses of the drugs, different conditioning regimens, and different restriction states. Here we have attempted to unify these variables by comparing the suppressive effects of a range of doses of morphine, cocaine, and LiCl on intake of a saccharin CS using a common regimen in non-restricted, food restricted, or water restricted male Sprague-Dawley rats. The results showed that, while the putatively aversive agent, LiCl, was effective in suppressing intake of the taste cue across nearly all doses, regardless of restriction state, the suppressive effects of both morphine and cocaine were greatly reduced when evaluated in either food or water restricted rats. Greater sensitivity to drug was revealed, at very low doses, when testing occurred in the absence of need (i.e., when the rats were non-restricted). Together, these results provide the first uniform and comprehensive analysis of the suppressive effects of morphine, cocaine, and LiCl as a function of dose and restriction state. In the present case, the suppressive effects of morphine and cocaine are found to differ from those of LiCl and, in some respects, from one another as well.