bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antidotes for Acute Cannabinoid Intoxication
Authors: Vemuri Kiran , Raghav Jimit , Makriyannis Alexandros 
Published: June 2021

The cannabis constituent Δ9‐tetrahydrocannabinol (THC), now being legalized for recreational and medical use, exerts its psychotropic effects (marijuana “high”) via CB1 receptor activation. Synthetic psychoactive cannabinoids (SPCs) such as JWH‐018, illegally sold as “designer drugs”, mimic the effects of THC, a CB1 partial agonist, with higher potency, acting via CB1 receptors. Although acute toxicity is benign for consuming high doses of THC, patients admitted to the ER may exhibit symptoms of hyperemesis, irritation of conjunctival and bronchial tissue, along with impaired cognition and motor skills. SPCs act as full CB1R agonists wherein clinical manifestations associated with their consumption include, neurotoxicity represented by several negative behavioral outcomes, and even death. While such adverse events may differ among patients, current adjunctive treatments include intubation, use of benzodiazepines, antipsychotics, antihistamines, adenosine, and hypnotics, along with i.v. saline, and supportive care. Currently, there are no effective treatments available for directly blocking and reversing the effects of cannabinoid poisoning. To address this critical public‐health issue, we have initiated a drug discovery program to develop antidotes that can counteract the intoxicating/adverse effects of CB1 activation and be therapeutically useful in an emergency setting. Although CB1 antagonists such as rimonabant appear to be promising pharmacotherapeutics, this class of drugs are known to produce nausea and unacceptable pro‐depressant effects. We have now discovered a novel class of potent, selective and brain penetrant CB1 antagonists, displaying better safety profiles as compared to rimonabant. In male CD‐1 mice, these novel CB1 antagonists represented by AM11503 (10 mg/kg, i.v), antagonized the hypothermic effect induced by THC (30 mg/kg, i.p.) and was equipotent to rimonabant (10 mg/kg, i.v.) in rapidly normalizing (≤ 60 min, post‐antagonist) the hypothermia induced by an acute dose of JWH‐018 (6 mg/kg, i.p). To mimic an ER‐like situation, antagonists were given i.v., 60 min post‐ JWH‐018, where the agonist effect is at its maximum. AM11503 (10 mg/kg, i.v or 18 mg/kg, i.p) displayed no side‐effects when compared to rimonabant and AM251, which, at similar doses, induced significant stereotypic behavior (pruritic responses). Remarkably, when tested for their ability to block the neurotoxic effects of SPCs, AM11503 (10 mg/kg, i.v.) fully blocked tremors and convulsions induced by “suprapharmacological” doses of JWH‐018 (18 mg/kg, i.p). Lead optimization efforts along with safety and efficacy studies are further underway. Here, we established proof‐of‐concept and present preliminary data towards developing the first specific antidote for cannabinoid intoxication.Support or Funding InformationSupported by NIDA/NIH grant DA045882 (to KV)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mechanism of Prenatal Cannabinoid Exposure Mediated Memory Loss in Adolescent Offspring: Opportunities for Identifying Therapeutic Target
Authors: Pinky Priyanka Das , Bloemer Jenna , Setti Sharay E , Heslin Ryan T , Smith Warren D , Du Yifeng , Dityatev Alexander , Reed Miranda N , Suppiramaniam Vishnu 
Published: June 2021

Cannabis use during pregnancy has increased by 62% from 2002 through 2014 and is now the most commonly used illicit drug during pregnancy, in part due to its ability to reduce morning sickness during pregnancy. Several studies have demonstrated that neural network activity which underlies typical cognitive and behavioral processes can be altered by prenatal cannabinoid exposure (PCE) leading to a long lasting effects on adult behavior. Here, we have investigated the impact of PCE in adolescent offspring in hippocampus dependent learning and memory via performing a series of behavioral, electrophysiological and immunochemical studies. An osmotic pump filled with either vehicle or the cannabinoid receptor full agonist WIN55, 212–2 (2 mg/kg body weight/day) was implanted subcutaneously in gestational day 4, which delivered the drug at a constant rate until the pups were born. Contextual Fear Conditioning and Morris Water Maze test were performed to investigate the hippocampus based memory revealing significant learning deficits in the PCE group. To investigate the mechanisms behind observed behavioral deficits, electrophysiological experiments were performed on acute hippocampal slices. Reduced long‐term potentiation and enhanced and long term depression were observed in the PCE group demonstrating impaired synaptic plasticity. To identify alterations in the downstream signaling cascade involved in glutamatergic neurotransmission, immunoblotting experiment was performed. Immunoblotting data showed increased Cannabinoid Receptor Type 1(CB1) expression along with reduced Neural Cell Adhesion Molecule (NCAM) expression. NCAM is important for maintaining proper neuronal connections, and it modulates NMDA mediated glutamatergic neurotransmission. Based on our data, we hypothesize that a reduction in NCAM may be responsible for the learning and memory deficits observed in PCE animals and can be used as a therapeutic target to ameliorate the cognitive deficits in PCE offspring.Support or Funding InformationNo Available Funding SourceThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cocaine increases the stimulation of dopamine release, at variance with atypical dopamine uptake inhibitors. New clues for the abuse liability of psychostimulants?
Authors: Keighron Jacqueline D , Bonaventura Jordi , Li Yang , Cao Jian Jing , DeMarco Emily M , Sandtner Walter , Michaelides Michael , Sitte Harald H. , Newman Amy H. , Tanda Gianluigi 
Published: June 2021

Dopamine uptake inhibitors (DUIs), such as cocaine and methylphenidate, are characterized by their ability to bind to the dopamine transporter (DAT) and prevent dopamine (DA) from being transported from the extracellular to the intracellular side of a neuronal terminal. It was originally posited that all DUIs would have the same effect on behavior, neurochemistry, and abuse liability in animal models as well as humans. However, a subset of these drugs, atypical DUIs (e.g. JHW007), have shown dissimilar profiles. While these DUIs have the same target and similar binding affinities to typical DUIs, their effects on behavior and neurochemistry are markedly different and predict little to no abuse liability in animal models.The goal of the present study was to determine how the neurochemistry of this atypical subset of DUIs differs from typical DUIs in mice. A pharmacological approach was taken utilizing Fast Scan Cyclic Voltammetry (FSCV), for real time monitoring of DA release and reuptake from the terminal; Fiber Photometry (FP), for real time monitoring of Ca2+ fluctuations in the terminal; and Electrophysiological (Ephys) ligand displacement experiments, to characterize the interactions between DUIs and the DAT.FSCV data indicate that cocaine and other abused DUIs have effects on both DA reuptake (by binding to the DAT and blocking DA transport) and stimulated release (causing an increase in mobilization of DA vesicles). In contrast the atypical DUIs, such as JHW007, have similar effects on DA reuptake, but do not cause a significant increase in stimulated DA release. FP experiments show how this difference influences Ca2+ fluctuations. JHW007 has a greater dose‐dependent impact on the influx of Ca2+ following stimulation than cocaine. Further Ephys experiments indicate a correlation between DA release and the off‐rate for each DUI, while no significant correlation was found for DA release and DAT affinity. Our data suggest 1) the dopaminergic effects of cocaine and other typical DUIs are not solely related to their ability to block the DAT; 2) DAT affinity alone is not an indicator for abuse liability; 3) the effect of the DUIs on the quaternary structure of the DAT and its downstream interactions may play an important role in the effects of typical DUIs related to abuse. Our preclinical results suggest that atypical DUIs can mitigate the effects of cocaine, leading to possible treatment candidates for psychostimulant use disorders in humans.Support or Funding InformationThis research was funded in part by the Medication Development Program, National Institute on Drug Abuse, IRP, NIH/DHHSThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
AAV‐7F9‐Fc Protects Against METH‐Induced Hyperthermia and Decreases in Blood Brain Barrier Tight Junction Expression in a Rodent Model
Authors: Bolden Chris , McLeroy‐Charles Kennede , Hay Charles E , Margaritas Paris , Peterson Eric 
Published: June 2021

Methamphetamine (METH) is a potent psychostimulant, with no FDA‐approved pharmacological treatment options. METH can adversely affect brain homeostasis by initiating dysfunction in the Blood Brain Barrier (BBB) and hyperthermia. The BBB is a semipermeable barrier composed of endothelial cells and pericytes along blood capillaries. These cells are held together by tight junction (TJ) proteins (e.g., claudin‐5, occludin, etc) which limit paracellular transport and help regulate body temperature. METH can decrease the expression of these TJ proteins and induce hyperthermia leading to BBB dysfunction. Thus, a novel therapy that could mitigate these effects is needed.A novel treatment under evaluation in our laboratory is an anti‐METH single chain variable antibody‐fusion antibody (7F9‐Fc) packaged into adeno‐associated virus (AAV8) particles. AAV‐7F9‐Fc works as a pharmacokinetic antagonist to slow and reduce entry of METH into the brain. AAV‐7F9‐Fc has achieved long‐term serum concentrations in mice (>7 months from a single dose) and has been shown to alter locomotor effects of METH in mice. This treatment could have clinical potential to attenuate the effects of METH during drug use and aid addiction recovery.We hypothesized that an AAV delivered anti‐METH antibody fragment (AAV‐7F9‐Fc) will mitigate METH‐induced hyperthermia and BBB dysfunction. To begin to test this hypothesis, we recorded baseline temperatures of mice 1 h before administration of either saline or METH (10 mg/kg). Male BALB/c mice were randomized into one of five groups (n=5/group); 1) Saline + Saline, 2) Saline + METH, 3) AAV‐7F9‐Fc + Saline, 4) AAV‐7F9‐Fc + METH, 5) AAV‐empty vector + saline. After administration of either Saline or METH, rectal temperatures were probed for 1 h, and brains were collected at the 3 h time point. Brains were dissected on dry ice into key dopaminergic regions such as the Frontal Cortex and Hippocampus. After dissection, proteins were extracted via RIPA buffer and analyzed for their expression by Western Blot. In Saline+METH treated mice, a 40% decrease in expression of Claudin‐5 and a 45% decrease in expression of Occludin (p < 0.05) were observed in the Frontal Cortex. These results were mitigated in AAV‐7F9‐Fc + METH mice in the Frontal Cortex and Hippocampus. These results indicate two important findings in an acute setting. This indicates that AAV‐7F9‐Fc has the potential to mitigate in an acute administration of METH, 1). the ability of METH to induce Hyperthermia, and 2). cause a reduction of critical tight junction protein expression in the CNS.Support or Funding InformationThis research was funded by NIH/NIDA:R01‐DA036600 & R01 DA036600‐03S1, NIH/NIGMS:T32‐GM106999, and R25GM083297.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Reward deficits in an animal model of compulsive eating
Authors: Cottone Pietro , Moore Catherine F , Leonard Michael , Miczek Klaus , Sabino Valentina 
Published: June 2021

BackgroundCompulsive eating, which affects an estimated 15 million adults in the US, can be conceptualized as overeating to relieve a negative emotional state. One hypothesized underlying mechanism is diminished reward. Overeating of palatable diets alter mesolimbic dopamine neurotransmission, likely contributing to the persistence of excessive intake. However, it is not yet known if overeating of palatable food alters reward sensitivity, and whether reward is differentially altered between phases of palatable food access and withdrawal.MethodsIn this study, we investigated sensitivity to d‐Amphetamine, a dopamine releaser, in rats undergoing ad libitum palatable diet cycling, a model of compulsive eating. In this model, animals receive 2 days of access to a highly palatable, chocolate flavored, high‐sucrose diet (‘P Phase’) followed by 5 days of access to a control chow diet (‘C Phase’). In all experiments, we investigated d‐ Amphetamine sensitivity during access to the palatable diet (‘P Phase’) as well as when animals were withdrawn from palatable food (‘C Phase’). To test behavioral and molecular response to d‐ Amphetamine, we used a locomotor response assay, conditioned place preference test, intra‐cranial self‐stimulation paradigm, and in vivo microdialysis. Finally, we tested oral self‐administration of d‐Amphetamine in the home cage. To examine dopaminergic signaling targets, gene expression of dopamine transporter (DAT) and tyrosine hydroxylase (TH) in the ventral tegmental area (VTA) were determined using quantitative polymerase chain reaction (qPCR).ResultsDuring the P phase, diet alternated rats showed reduced sensitivity to d‐Amphetamine's stimulating (locomotor assay), rewarding (CPP), and reward‐enhancing effects (ICSS). In diet cycled animals, d‐Amphetamine‐evoked dopamine efflux was blunted. Chow/Palatable rats had higher self‐administration of d‐Amphetamine during the P Phase. Levels of DAT mRNA were increased in the VTA in Chow/Palatable rats during the P Phase.ConclusionsThese results indicate that a history of overconsumption of palatable food causes reward deficits and associated alterations in the mesolimbic pathway that led to increased self‐administration of d‐ Amphetamine.Support or Funding InformationR01‐030425, F31‐DA044664This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Icariin improves the cognitive function of APP/PS1 mice via suppressing ER stress
Authors: Li Fei , Zhang Yangyang , Shi Jingshan , Gong Qihai 
Published: June 2021

Alzheimer's disease (AD) is a progressive neurodegenerative disorder including many cognitive processes decline and neurons loss. Massive misfolded or unfolded proteins gathered in endoplasmic reticulum (ER) can disturb ER function and induce ER stress, which results in neuronal death in AD. Icariin (ICA), extracted from Chinese herb, has a wide range of neuron protection and has been researched in AD treatment. The current study was focusing on the effect of ICA on ER stress in AD animal model. We found that ICA improved 12‐month aged APP/PS1 mice behavior performance, reduced Aβ1‐40/42 deposition in hippocampal region. We also observed the APP‐ Aβ pathway, and found ICA decreased the protein expression of APP and BACE1, as well as increased the level of ADAM10, indicating that ICA inhibited aberrant APP‐Aβ metabolism by reducing APP and β‐secretase. In order to observe the effect of ICA on ER stress in AD condition, we tested the ER stress marker GRP78 and the results showed that ICA decreased the protein and mRNA level of GRP78 in APP/PS1 mice hippocampus. We further observed the main three branches of ER stress, the phosphorylation level PERK and eIF2α has been decreased in APP/PS1 mice after ICA treatment, the downstream factors of eIF2α (ATF4 and CHOP) were also decreased, but IRE1 phosphorylation and ATF6 were not affected in ICA treated mice. This suggested that ICA maybe inhibit ER stress through altering the PERK/eIF2α pathway. Furthermore, we found the neuron apoptosis in APP/PS1 mice were ameliorated after ICA treatment, the level of cleaved‐casepase‐3, ‐9, ‐12, as well as Bax/Bcl‐2 ratio were dramatically decreased in ICA treated APP/PS1 mice. These results demonstrated that ICA could serve as a potent ER stress inhibitor in AD research.Support or Funding InformationThis work was supported by the National Natural Science Foundation of China (Grant No. 81560594); Guizhou Province International Science and Technology cooperation project (Qiankehewai G [2014]7011).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Fear Extinction and Myocardial Infarction on Corticotropin Releasing Hormone Receptor 1 mRNA in Rostral Locus Coeruleus of Rats
Authors: Rose Deborah , Kuckertz Anne , Scrogin Karie 
Published: June 2021

We found that 8 weeks after myocardial infarction (MI) female, but not male rats showed deficits in extinction of conditioned fear (P<0.05). Small hairpin RNA‐mediated knockdown of corticotropin releasing hormone receptor 1 (CRF1) around the rostral locus coeruleus (rLC) reversed deficits in fear extinction. Female and male rats subjected to MI and sham knockdown showed reduced CRF1 mRNA levels 24 hours after extinction training compared to sham MI/sham knockdown controls (P<0.01). CRF1 mRNA levels in the rLC region were directly related to the extent of freezing observed during extinction trials in rats subjected to sham knockdown (P<0.05). These findings suggest either that exposure to feared cues during extinction contributed to acute reductions in CRF1 mRNA in the rLC region, or that chronic reductions in CRF1 mRNA levels resulted from MI. Here, we determined if CRF1 mRNA levels in the rLC region were reduced after MI in male rats. Fresh frozen brains of rats exposed to coronary artery ligation (n=4) or sham surgery (n=3) 8 weeks prior to euthanasia were processed for fluorescent in situ hybridization. Brain sections containing the rLC were processed for detection of CRF1‐, vesicular glutamate transporter 2‐ (VGLUT2, a glutamatergic cell marker) and glutamate decarboxylase 2 mRNA (GAD2, a GABAergic cell marker). Assessments from the medial parabrachial nucleus to the lateral border of the posterodorsal tegmental nucleus and ventral border of Barrington's nucleus were performed. Cells were counted bilaterally and averaged within rat. No differences in CRF1 mRNA were observed between groups. A small number of CRF1 mRNA positive cells (39±6) close to the rLC did not show VGLUT2 or GAD2 mRNA label. Numbers of GAD2‐ (90±15) and VGLUT2 mRNA‐positive cells (365±30) did not differ between groups. CRF1‐ and VGLUT2 mRNA‐positive cells were found in high density in the medial parabrachial nucleus and in the lateral dorsal tegmental nucleus. CRF1‐ and GAD2 mRNA‐positive cells were found medially, near the posterodorsal tegmental nucleus. The percent of VGLUT2 mRNA‐positive cells (72±4) that co‐expressed CRF1 mRNA was greater than the percent of GAD2 mRNA‐positive cells that co‐expressed CRF1 mRNA (60±5, p<0.001). The integrated density of CRF1 mRNA fluorescence was greater in VGLUT2 mRNA‐positive cells than in GAD2 mRNA‐positive cells (P<0.05). The data indicate that CRF1 mRNA is expressed primarily by glutamatergic cells in the rLC region of the male rat and that reductions in CRF1 mRNA levels observed in the rLC region 24 hours after extinction in MI rats are likely due to stress responses during fear extinction rather than to chronic effects of MI.Support or Funding InformationCardiovascular Research Institute, Loyola University ChicagoThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigating the Role of Serotonin Transporter, Plasma Membrane Monoamine Transporter, and Organic Cation Transporter 3 in the Antidepressant‐like Effects of Ketamine
Authors: Bowman Melodi Ann , Vitela Melissa , Owens W. Anthony , Koek Wouter , Daws Lynette C. 
Published: June 2021

Twenty percent of adults are diagnosed with major depressive disorder, which is typically treated with selective serotonin reuptake inhibitors (SSRIs). However, SSRIs take approximately six weeks to produce therapeutic effects. Recently, low doses of ketamine, a noncompetitive N‐methyl‐ D‐aspartate (NMDA) receptor antagonist, have been shown to produce rapid and long‐lasting antidepressant effects. Studies that have begun to examine the mechanisms underlying these effects have focused on intracellular changes mediated by NMDA and/or AMPA receptors. However, the traditional view of the etiology of depression involves the need for an increase in extracellular serotonin to regulate mood. Surprisingly, there appears to be little research into the effects of ketamine on serotonin. One study found an increase in extracellular serotonin following ketamine, consistent with results from another study showing that ketamine inhibited serotonin uptake. Together, these findings suggest a role for serotonin in the antidepressant‐like effects of ketamine, and putatively one involving transporters capable of serotonin uptake. The serotonin transporter (SERT) is an obvious candidate, but recent evidence points to the plasma membrane monoamine transporter (PMAT) and organic cation transporter (OCT3) as important players in serotonin uptake as well. In order to investigate a role for these transporters in the action of ketamine to inhibit serotonin uptake, we used in vivo chronoamperometry to measure serotonin clearance from extracellular fluid of CA3 region of the hippocampus. In wildtype mice, ketamine robustly inhibited serotonin clearance. Ongoing chronoamperometry studies are using knockout lines of SERT, PMAT and OCT3 mice to parse apart their roles in the action of ketamine to inhibit serotonin clearance. Parallel studies using these knockout (KO) lines in the forced swim test support the involvement of all three transporters in the antidepressant‐like effect of ketamine. As expected, ketamine (32 mg/kg) produced robust antidepressant‐like effects in wildtype mice, but in marked contrast, was without effect in SERT KO, PMAT KO, and OCT3 KO mice. Ketamine did not influence locomotor activity in SERT KO and wildtype mice, ruling out differences in the action of ketamine on general activity accounting for the contrasting effect of ketamine in the FST between these genotypes. Experiments assessing the locomotor actions of ketamine in PMAT KO and OCT3 KO mice are ongoing. Together, these data suggest that blockade of SERT, PMAT, and/or OCT3 may be important for the antidepressant‐like effects of ketamine.Support or Funding InformationR01 MH093320R01 MH106978T32 NS082145This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.