The role of CART peptide in learning and memory: A potential therapeutic target in memory-related disorders.

Cocaine and amphetamine-regulated transcript (CART) mRNA and peptide are vastly expressed in both cortical and subcortical brain areas and are involved in critical cognitive functions. CART peptide (CARTp), described in reward-related brain structures, regulates drug-induced learning and memory, and its role appears specific to psychostimulants. However, many other drugs of abuse, such as alcohol, opiates, nicotine, and caffeine, have been shown to alter the expression levels of CART mRNA and peptides in brain structures directly or indirectly associated with learning and memory processes. However, the number of studies demonstrating the contribution of CARTp in learning and memory is still minimal. Notably, the exact cellular and molecular mechanisms underlying CARTp effects are still unknown. The discoveries that CARTp effects are mediated through a putative G-protein coupled receptor and activation of cellular signaling cascades via NMDA receptor-coupled ERK have enhanced our knowledge about the action of this neuropeptide and allowed us to comprehend better CARTp exact cellular/molecular mechanisms that could mediate drug-induced changes in learning and memory functions. Unfortunately, these efforts have been impeded by the lack of suitable and specific CARTp receptor antagonists. In this review, following a short introduction about CARTp, we report on current knowledge about CART's roles in learning and memory processes and its recently described role in memory-related neurological disorders. We will also discuss the importance of further investigating how CARTp interacts with its receptor(s) and other neurotransmitter systems to influence learning and memory functions. This topic is sure to intrigue and motivate further exploration in the field of neuroscience.

Protective Role of Ellagic Acid Against Ethanol-Induced Neurodevelopmental Disorders in Newborn Male Rats: Insights into Maintenance of Mitochondrial Function and Inhibition of Oxidative Stress.

OBJECTIVE: Ellagic acid (EA) exerts, neuroprotective, mitoprotective, anti-oxidative and anti-inflammatory effects. We evaluated protective effect of EA on ethanol-induced fetal alcohol spectrum disorders (FASD). METHODS: A total of 35 newborn male rats were used, divided into five groups, including; control (normal saline), ethanol (5.25 g/kg per day), ethanol (5.25 g/kg per day) + EA (10 mg/kg), ethanol (5.25 g/kg per day) + EA (20 mg/kg) and ethanol (5.25 g/kg per day) + EA (40 mg/kg). Thirty-six days after birth behavioral tests (Morris water maze and Elevated Plus Maze), tumor necrosis factor-α (TNF-α) levels, oxidative markers (malondialdehyde, glutathione and superoxide dismutase), mitochondrial examination such as succinate dehydrogenases (SDH) activity, mitochondrial swelling, mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) formation were analyzed. RESULTS: The results revealed that ethanol exposure adversely affected cognitive and mitochondrial functions and as well as induced oxidative stress and inflammation in brain tissue. However, EA (20 and 40 mg/kg) administration effectively prevented the toxic effects of ethanol in FASD model. CONCLUSIONS: These findings demonstrate that ethanol application significantly impairs the brain development via mitochondrial dysfunction and induction of oxidative stress. These data indicate that EA might be a useful compound for prevention of alcohol-induced FASD.

Hesperidin Protects Alcohol-Induced Mitochondrial Abnormalities via the Inhibition of Oxidative Stress and MPT Pore Opening in Newborn Male Rats as a Fetal Alcohol Syndrome Model.

OBJECTIVE: Prenatal alcohol exposure causes fetal developmental abnormalities via mitochondrial dysfunction, reactive oxygen species (ROS) formation, and oxidative stress. Therefore, we aimed to investigate the potential of hesperidin as a mitochondrial protective and antioxidative agent in newborn male rats as a model for fetal alcohol syndrome (FAS). METHOD: Newborn male rats were divided randomly into five groups: a sham group (receiving 27.8 ml/ kg milk solution, orally), an ethanol group (5.25 g/kg in milk solution, orally, 2-10 days after birth), an ethanol + hesperidin group (25 mg/kg/ day orally), an ethanol + hesperidin group (50 mg/kg/day orally), and an ethanol + hesperidin group (100 mg/kg/day orally). Thirty-six days after birth, newborn male rats were sacrificed and brain mitochondria were isolated using differential centrifugation. Mitochondrial toxicity biomarkers of succinate dehydrogenase (SDH) activity, mitochondrial swelling, mitochondrial membrane potential (MMP), and ROS were measured. RESULTS: Offspring neonatally exposed to ethanol showed a significant reduction in SDH activity, mitochondrial swelling, MMP collapse, induction of ROS formation, and lipid peroxidation in isolated mitochondria. Oral administration of hesperidin restored SDH activity, improved MMP collapse and mitochondrial swelling, and reduced ROS formation. CONCLUSIONS: This study demonstrates that hesperidin exerts a potent protective effect against alcohol-induced mitochondrial toxicity in the FAS model. Moreover, these findings indicate that hesperidin might be a useful compound for prevention of alcohol-induced fetal developmental abnormalities during pregnancy.

Validity of the Persian version of the Munich ChronoType Questionnaire (MCTQIR).

PURPOSE: Currently, the most useful questionnaire for determining the chronotype is the Munich ChronoType Questionnaire (MCTQ). It determines chronotype based on sleep-wake behavior on workdays and work-free days and uses the mid-sleep time on free days (MSF), corrected for accumulated sleep debt over the work week to classify chronotype (MSFsc). Our study aimed at validating Persian version of the MCTQ. METHODS: Participants completed the Persian version of the MCTQ, reduced Morningness-Eveningness Questionnaire (rMEQ), Epworth sleepiness scale (ESS), and demographic characteristics. RESULTS: The study sample was comprised of 250 Iranian men and women (mean age 34 years, age range 30 to 58 years, 113 men). Results showed that there is a significant negative relationship between the components of MCTQ and rMEQ, and the strongest relationship related to the MSF index. The results also showed that MCTQ parameters were not significantly related to the ESS scale. Cut-off values of MSFsc for chronotype categories were determined by inter-quartile range (Q25% = 3.44, Q50% = 4.68, and Q75% = 5.75). CONCLUSIONS: Like the MCTQ in other languages, the Persian version of the MCTQ is a reliable tool for assessing chronotype.

Validation and Psychological Properties of the Persian Version of DSM 5 Yale Food Addiction Scale 2.0 (PYFAS 2.0) in Non-clinical Population.

Background: The Yale Food Addiction Scale version 2.0 (YFAS 2.0) is used for the assessment of food addiction (FA). This research intended to evaluate the validity of the Persian translation of the questionnaire and to investigate the psychological properties and the association between FA and anthropometric indices. Methods: In a sample of 473 nonclinical participants, FA, binge eating, and objectively measured anthropometric indices were assessed. Internal consistency, convergent, and validity of the PYFAS 2.0 were examined. Also, the factor structure (confirmatory factor analysis following the 11 diagnostic indicators in addition to the significant distress) and the construct of the scale were evaluated. Findings: The frequencies of mild, moderate, and severe FA based on PYFAS 2.0 were 0.2%, 10%, and 5.5%, respectively. The findings supported a one-factor structure. The confirmatory factor analysis revealed a good construct validity (RMSEA=0.043, χ2=76.38, df=41, χ2 (CMIN)/df=1.862, GFI=0.975, AGFI=0.957, IFI=0.986, RFI=0.958, ECVI=0.319, TLI=0.978). For both the diagnostic and symptom count versions, the PYFAS 2.0 presented acceptable internal consistency (IC) (Kuder-Richardson 20=0.99 and McDonald omega=0.91). Conclusion: The PYFAS 2.0 was a psychometrically sound instrument in an Iranian non-clinical population. This questionnaire can be used to study FA in Persian non-clinical populations. Future research should study the psychometric characteristics of this scale in high-risk groups.

Erythropoietin improve spatial memory impairment following methamphetamine neurotoxicity by inhibition of apoptosis, oxidative stress and neuroinflammation in CA1 area of hippocampus.

OBJECTIVE: Methamphetamine (METH) is one of the most widely used addictive drugs, and addiction to it is on the rise all over the world. METH abuse has long-term damaging effects that reduce memory and impair cognitive functions. According to studies, the observed effects are strongly related to the nerve cell damage caused by METH, which leads to neurotoxicity. Some of these intra-neuronal events include dopamine oxidation, excitotoxicity, and oxidative stress. Erythropoietin (EPO) is a hormone produced primarily by the kidneys and, in small quantities, by the liver. Studies have shown that EPO exhibits considerable neuroprotective effects. This study aimed to investigate the protective effects of EPO on METH neurotoxicity. METHODS: Initially, 48 male Wistar rats, weighing 250-300 g, were randomly assigned to four groups: control (n = 12), METH (n = 12), and METH+EPO (2500, 5000 IU/kg/IP- n = 12). METH was injected intraperitoneally at a dose of 40 mg per kg of body weight (four injections of 10 mg every two hours) to induce neurotoxicity. EPO was injected at doses of 2500 and 5000 IU/kg seven days after the last METH administration (ip). Morris water maze test was performed following EPO injection (1 day after the last dose) to assess spatial memory. The brains were removed after the behavioral test, biochemical evaluations and immunohistochemistry (caspase-3 and GFAP) was performed. RESULTS: The results showed that EPO treatment significantly improved spatial memory impairment (P < 0.01), compared to the METH group, EPO was a significant reduction in malondialdehyde and TNF-α (P < 0.01), as well as an increase in superoxide dismutase (P < 0.05) and glutathione-PX (P < 0.01). Furthermore, EPO treatment significantly reduced the number of GFAP positive cells (P < 0.01) and caspase 3 (P < 0.001) in the hippocampus (CA1 region). CONCLUSIONS: The study findings suggested that EPO may have great neuroprotective effects on METH neurotoxicity due to its anti-inflammatory, antioxidant, and antiapoptotic properties.

Neurological manifestations of coronavirus infections, before and after COVID-19: a review of animal studies.

Coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This virus, which was first identified in December 2019 in China, has resulted in a yet ongoing viral pandemic. Coronaviridae could potentially cause several disorders in a wide range of hosts such as birds and mammals. Although infections caused by this family of viruses are predominantly limited to the respiratory tract, Betacoronaviruses are potentially able to invade the central nervous system (CNS) as well as many other organs, thereby inducing neurological damage ranging from mild to lethal in both animals and humans. Over the past two decades, three novel CoVs, SARS-CoV-1, MERS-CoV, and SARS-CoV-2, emerging from animal reservoirs have exhibited neurotropic properties causing severe and even fatal neurological diseases. The pathobiology of these neuroinvasive viruses has yet to be fully known. Both clinical features of the previous CoV epidemics (SARS-CoV-1 and MERS-CoV) and lessons from animal models used in studying neurotropic CoVs, especially SARS and MERS, constitute beneficial tools in comprehending the exact mechanisms of virus implantation and in illustrating pathogenesis and virus dissemination pathways in the CNS. Here, we review the animal research which assessed CNS infections with previous more studied neurotropic CoVs to demonstrate how experimental studies with appliable animal models can provide scientists with a roadmap in the CNS impacts of SARS-CoV-2. Indeed, animal studies can finally help us discover the underlying mechanisms of damage to the nervous system in COVID-19 patients and find novel therapeutic agents in order to reduce mortality and morbidity associated with neurological complications of SARS-CoV-2 infection.

Circadian rhythm influences naloxone induced morphine withdrawal and neuronal activity of lateral paragigantocellularis nucleus.

Investigations have shown that the circadian rhythm can affect the mechanisms associated with drug dependence. In this regard, we sought to assess the negative consequence of morphine withdrawal syndrome on conditioned place aversion (CPA) and lateral paragigantocellularis (LPGi) neuronal activity in morphine-dependent rats during light (8:00-12:00) and dark (20:00-24:00) cycles. Male Wistar rats (250-300 g) were received 10 mg/kg morphine or its vehicle (Saline, 2 mL/kg/12 h, s.c.) in 13 consecutive days for behavioral assessment tests. Then, naloxone-induced conditioned place aversion and physical signs of withdrawal syndrome were evaluated during light and dark cycles. In contrast to the behavioral part, we performed in vivo extracellular single-unit recording for investigating the neural response of LPGi to naloxone in morphine-dependent rats on day 10 of morphine/saline exposure. Results showed that naloxone induced conditioned place aversion in both light and dark cycles, but the CPA score during the light cycle was larger. Moreover, the intensity of physical signs of morphine withdrawal syndrome was more severe during the light cycle (rest phase) compare to the dark one. In electrophysiological experiments, results indicated that naloxone evoked both excitatory and inhibitory responses in LPGi neurons and the incremental effect of naloxone on LPGi activity was stronger in the light cycle. Also, the neurons with the excitatory response exhibited higher baseline activity in the dark cycle, but the neurons with the inhibitory response showed higher baseline activity in the light cycle. Interestingly, the baseline firing rate of neurons recorded in the light cycle was significantly different in response (excitatory/inhibitory) -dependent manner. We concluded that naloxone-induced changes in LPGi cellular activity and behaviors of morphine-dependent rats can be affected by circadian rhythm and the internal clock.

Aqueous-alcoholic Ferulla extract reduces memory impairments in rats exposed to cadmium chloride.

INTRODUCTION: Cadmium (Cd) is the most dangerous heavy metal that is becoming more widespread in nature as a result of industrial activities. One of the toxic effects of Cd on the body is its neurological effect. The mechanism of these effects has been attributed to the induction of oxidative stress. Ferulla plant has antioxidant properties. In the present study, the aim was to reduce the toxic effects of Cd on memory impairment in rats by through the consumption of Ferulla extract. MATERIALS & METHOD: Rats were randomly divided into five groups of six: (1) control group, (2) 300 µM cadmium exposure group, and three treatment groups with doses of (3) 100, (4) 300, and (5) 600 mg/kg.BW of F. Ferulla extract after Cd exposure. To induce neurotoxicity, Cd was daily injected peritoneally at a concentration of 300 µM in 1 ml of normal saline for a week. Next, for 3 weeks, the Cd group received 1 ml of normal peritoneal saline, and the treatment groups received F. Ferulla extract at concentrations of 100, 300, and 600 mg/kg.BW in 1 ml of normal saline daily for a week. At the end of the treatment period, a water maze was used to assess memory disorders. Malondialdehyde (MDA), glutathione concentration (GSH), and glutathione peroxidase (GPX) activity in nerve tissue were also measured. Morris water maze was also performed after intervention. RESULTS: Cd-induced neurotoxicity was shown in Cd groups. MDA, GSH, and GPX have a significant difference in comparison between the Cd and 300, 600 treated groups. MDA has a significant increase (p < 0.05), and GSH and GPX have a significant decrease (p < 0.05). The results of the Morris water maze showed that the Cd group spent either 300 or 600 more distances and time to find a place to escape, which was significant (p < 0.05) CONCLUSION: Cd exposure can induce neurotoxicity and disrupt learning and memory. On the other hand, Ferulla extract can improve learning and memory in Cd-induced neurotoxicity model via induced antioxidant defense system.

Cyanocobalamin improves memory impairment via inhibition of necrosis and apoptosis of hippocampal cell death after transient global ischemia/reperfusion.

OBJECTIVES: Brain ischemia/reperfusion (I/R) causes irreversible damage, particularly in the hippocampus. Cyanocobalamin (CNCbl) is known to be crucial for the proper operation of the nervous system. Vitamin B12 has been demonstrated to exert antioxidant effects via direct and indirect mechanisms. It can also protect cortical neurons against glutamate cytotoxicity. This research was conducted to examine CNCbl protection against neuronal cell death in the rat hippocampal region following transient cerebral ischemia. MATERIALS AND METHODS: In this experiment, 48 male Wistar rats were selected, which were randomly divided into four groups (n=12 in each group): sham, ischemia/reperfusion, ischemia/reperfusion + CNCbl 200 and 400 (µg/kg). By occlusion of both common carotids, ischemia induction was performed within 20 min. CNCbl at the doses of 200 and 400 µg/kg was injected (IP) at the start of the reperfusion, 24 and 48 hr following reperfusion. The spatial memory was assessed 7 days following ischemia through the Morris water maze test. Antioxidant enzymes, apoptosis, and necrosis were measured after behavioral tests. RESULTS: CNCbl significantly improved spatial memory impairments (P<0.05), also CNCbl therapy significantly increased both glutathione (P<0.01) and superoxide dismutase (P<0.05) and reduced malondialdehyde (P<0.01) and TNF-α (P<0.05) in comparison with the ischemia group. In addition, CNCbl significantly decreased both apoptosis and necrosis in the hippocampus CA1 (P<0.01). CONCLUSION: CNCbl improves memory impairment following ischemia injury by decreasing neuronal cell death via its antioxidant properties.

Advances in immunotherapy for COVID-19: A comprehensive review.

COVID-19 is an acute respiratory syndrome caused by SARS-COV-2 which has now become a huge pandemic worldwide. The immunopathogenesis of COVID-19 has been established that increased serum levels of C-reactive protein (CRP), interleukin-6 (IL-6), and reduction of the CD4+ and the CD8+ T lymphocyte populations, are the most reported immunological findings in these patients. High levels of other inflammatory cytokines and chemokines such as IL-2 and IL-8 with an increased number of neutrophils and eosinophils may induce immune abnormalities in patients with COVID-19. There is growing evidence to obtain a deeper understanding of the immunopathogenesis of COVID-19 which will lay the foundation for the development of new potential therapies. However, specific and non-specific immunotherapies such as convalescent plasma (CP) are widely performed to treat patients with severe COVID-19, there is no definitive evidence to suggest the effectiveness of these treatments. Hence, this review aimed to highlight the current and most recent studies to identify the new immunotherapeutics for COVID-19 disease.

Apelin-13 attenuates spatial memory impairment by anti-oxidative, anti-apoptosis, and anti-inflammatory mechanism against ethanol neurotoxicity in the neonatal rat hippocampus.

It has been shown that alcohol consumption by pregnant women can have detrimental effects on the developing fetus and lead to fetal alcohol spectrum disorders (FASD). Exposure to alcohol in rat pups during this period causes long-term changes in the structure of the animal's hippocampus, leading to impaired hippocampal-related brain functions such as navigation tasks and spatial memory. Apelin-13, a principal neuropeptide with inhibitory effects on neuroinflammation and brain oxidative stress production, has beneficial properties on memory impairment and neuronal injury. The protective effects of apelin-13 have been evaluated on ethanol-related neurotoxicity in the hippocampus of rat pups. Rat pups from 2 until 10 postnatal day, similar to the third trimester of pregnancy in humans, were intubated total daily dose of ethanol (5/27 g/kg/day). Immediately after intubation, 25 and 50 µg/ kg of apelin-13 was injected subcutaneously. By using Morris water maze task, the hippocampus- dependent memory and spatial learning were evaluated 36 days after birth. Then, Immunohistochemical staining was done to determine the levels of GFAP and caspase-3. ELISA assay was also performed to measure both TNF-α and antioxidant enzymes levels. The current study demonstrates that administration of apelin-13 attenuates spatial memory impairment significantly (P < 0.001). After ethanol neurotoxicity, apelin-13 could also increase the catalase level (P < 0.001), activity of total superoxide dismutase as well as glutathione concentration noticeably (P < 0.05). Other impacts of it could be mentioned as attenuating TNF-α production and also preventing lipid peroxidation (P < 0.001). In addition, the results showed that the level of GFAP as a neuroinflammation factor and the number of active caspase-3 positive cells can be decreased by apelin-13 (P < 0.01). Regarding the protective effects of apelin-13 against ethanol-induced neurotoxicity, it is a promising therapeutic choice for FASD; but more studies are needed.

Hydrogen sulfide protects hippocampal CA1 neurons against lead mediated neuronal damage via reduction oxidative stress in male rats.

H2S plays vital roles in modulation brain function. It is associated with antioxidant and anti-inflammatory properties. We assessed the H2S impact on spatial learning and memory deficit and cell death due to lead exposure, and probable mechanisms of action. The 36 male Wistar rats that (200-220 g), were in random assigned to 3 groups, control group (12 rats), lead acetate group (12 rats), and lead acetate +H2S groups (NaHS as a H2S donor; 5/6 mg/kg; 12 rats). Administration of lead to rats was performed through acute lead poisoning (25 mg/kg of lead acetate, IP through 3 days). Using male Morris water maze, their spatial learning and memory function were measured. We carried out ELISA method to calculate TNF-α and antioxidant enzymes level. Immunohistochemical staining was applied for evaluating the caspase-3 expression levels. Treatment with H2S improved learning and memory impairment in Pb-exposed rats (P<0.05). H2S treatment suppressed Pb-related apoptosis in the hippocampal CA1 subfield (P<0.01). Also, the TNF-α over-expression in the CA1 region of hippocampus due to lead exposure showed a significant reduction (P<0.05) after administrating H2S. Simultaneously, H2S treatment reduced the MDA levels, enhanced SOD, GSH level than the Pb-exposed group in hippocampus (P<0.05). H2S was able to significantly improve Pb-related spatial learning and memory deficit, and neuronal cell death in the CA1 region of hippocampus in the male rats at least partly by reducing oxidative stress and TNF.

Improving sciatic nerve regeneration by using alginate/chitosan hydrogel containing berberine.

Peripheral nerve injuries are the common results of trauma that lead to pain and handicap in patients. Berberine due to its properties like antibiotic, immunostimulant, antitumor, antimotility, and positive effect on neurological disorders can be used to enhance peripheral nerve injuries. In this study, alginate/chitosan hydrogel containing different concentrations of berberine (0, 0.1, 1, 10% (w/v)) was created, evaluated, and applied as a scaffold for sciatic nerve regeneration. To prepare hydrogel, sodium alginate was dissolved in distilled water and cross-linked with CaCl2, and chitosan was dissolved in acetic acid and cross-linked with ß-glycerol phosphate. The structure, release, swelling, weight loss, cytocompatibility, and hemocompatibility of the prepared hydrogels were assessed. The sciatic nerve crush was created in rats and fabricated hydrogels were injected, and functional analysis was used to evaluate their effectiveness. The results of physical characterization of the hydrogel indicated that the initial average pore size was about 39 µm and about 70% of the main weight of hydrogels was lost after incubation for 21 days and hemocompatibility of hydrogels was also confirmed. The MTT assay showed the cytocompatiblity of hydrogels and also indicated that berberine has dose-dependence effect on cell proliferation. The in vivo results showed the positive effect of berberine especially the hydrogel contained 1% of berberine on regeneration of sciatic nerve. Based on this study, Alg/Chit hydrogel can be applied as a treatment to heal peripheral nerve injuries. Graphical abstract.

The effect of microinjection of CART 55-102 into the nucleus accumbens shell on morphine-induced conditioned place preference in rats: Involvement of the NMDA receptor.

The addictive properties of opioids may be mediated to some extent by cocaine-and amphetamine-regulated transcript (CART) in the reward pathway. Moreover, some claims CART interacts with the glutamate system. Here, we evaluated whether intra-nucleus accumbens (NAc) shell infusions of CART induces Conditioned Place Preference (CPP) or Conditioned Place Aversion (CPA) and affects morphine reward. We also measured NR1 subunit expressions of the N-methyl-d-aspartate (NMDA) receptor in various parts of the reward pathway (NAc, prefrontal cortex and hippocampus) after conditioning tests. Animals with bilateral intra-NAc shell cannulas were place-conditioned with several doses of subcutaneous morphine prior to intra-NAc shell infusions of artificial cerebrospinal fluid (aCSF). Immunohistochemistry (IHC) showed a dose-dependent increase in the NR1 expression in all examined parts. When rats were conditioned with intra-NAc shell infusions of CART, CPP and CPA induced with 2.5 and 5 µg/side respectively and IHC showed NR1elevation with 2.5 and reduction with 5 µg/side in all areas. Sub-rewarding dose of CART administration (1.25 µg/side) prior to sub-rewarding dose of morphine (2.5 mg/kg) induced CPP and NR1 increased in all examined tissues in IHC. However, infusion of an aversive dose of CART (5 µg/side) prior to the rewarding dose of morphine (5 mg/kg) produced neither CPP nor CPA and NR1 in the NAc and hippocampus decreased significantly. It seems that the rewarding or aversive effects of intra-NAc shell CART and its facilitating or inhibiting effects on morphine reward are dose-dependent. Additionally, NMDA may be closely involved in the affective properties of opioids and CART in the reward pathway.

Evaluation of the CART peptide expression in morphine sensitization in male rats.

The importance of Cocaine- and amphetamine-regulated transcript (CART) peptide in reinforcing effects of addictive drugs specially alcohol and psychostimulants has been stablished. Involvement of CART peptide in rewarding effects of opioids in brain has recently been reported. Here we have studied the expression of CART mRNA and peptide in the reward pathway in morphine-induced sensitization phenomenon and also evaluated the peptide level fluctuations in CSF and plasma. Male Wistar rats received 7-day morphine injection (20mg/kg) and then after a 7-day washout period, a challenge dose of 10mg/kg morphine was administered and locomotor activity and oral stereotypical behaviors were recorded. Besides, the expression level of CART mRNA and peptide in four important areas of the mesocorticolimbic reward pathway including nucleus accumbens, striatum, prefrontal cortex, and hippocampus were measured by real-time PCR and western blotting, respectively. The level of the peptide in CSF and plasma was measured by Elisa method. The expression level of CART mRNA and protein in brain regions and also the peptide level in CSF and plasma were significantly down-regulated after 7-day morphine administration. These reduced levels returned to nearly normal rates after 7-day wash-out period. Administration of morphine challenge dose led to significant upregulation of CART gene expression (both mRNA and peptide) in the brain, and elevation of peptide level in CSF and plasma in morphine-sensitized rats. It can be concluded that CART is released in the framework of reward pathway and may serve as an important neurotransmitter in the process of morphine dependence and sensitization.

Evaluation of CART peptide level in rat plasma and CSF: Possible role as a biomarker in opioid addiction.

It has been shown previously that cocaine- and amphetamine-regulated transcript (CART) peptide has a modulatory role and homeostatic regulatory effect in motivation to and reward of the drugs of abuse specially psychostimulants. Recent data also showed that in addition to psychostimulants, CART is critically involved in the different stages of opioid addiction. Here we have evaluated the fluctuations in the level of CART peptide in plasma and CSF in different phases of opioid addiction to find out whether CART can serve as a suitable marker in opioid addiction studies. Male rats were randomly distributed in groups of control, acute low-dose (10mg/kg) morphine, acute high-dose morphine (80mg/kg), chronic escalating doses of morphine, withdrawal syndrome precipitated by administration of naloxone (1mg/kg), and abstinent after long-term drug-free maintenance of addicted animals. The level of CART peptide in CSF and plasma samples was measured by enzyme immunoassay. CART peptide concentration in the CSF and plasma was significantly elevated in acute high-dose morphine and withdrawal state animals and down-regulated in addicted rats. In abstinent group, CART peptide level was up-regulated in plasma but not in CSF samples. As the observed results are in agreement with data regarding the CART mRNA and protein expression in the brain reward pathway in opioid addiction phases, it may be suggested that evaluation of CART peptide level in CSF or plasma could be a suitable marker which reflects the rises and falls of the peptide concentration in brain in the development of opioid addiction.