bluuebunny/crossref_metadata_2025_split huggingface dataset:
Protective Effect of Propranolol and Nadolol on PTSD‐like Behavior in Rats
Authors: Zaidi Safiyya , Liu Hesong , Valdez Daniel , Kochi Camila , Atrooz Fatin , Salvi Ankita , Bond Richard , Salim Samina 
Published: November 2023

Traumatic experiences are reported to cause Post Traumatic Stress Disorder (PTSD). Benzodiazepines and Selective Serotonin Re‐Uptake Inhibitors are considered as standard treatments. Although clinically useful, chronic use causes drastic side effects. Thus, there remains a necessity for improving treatment and finding preventive pharmacological interventions. While promising evidence emerged with administration of b‐adrenergic receptor antagonist Propranolol and PTSD relief, methodological limitations of the studies and efficacy issues dampened the excitement. We believe it is premature to completely eliminate the role of b‐antagonists in PTSD. Using the social defeat paradigm, we examined the effect of two b‐adrenergic receptor antagonists Propranolol and Nadolol on PTSD‐like behavior in Sprague Dawley rats. Propranolol was administered in drinking water at ~18 mg/rat/day for 36 days. Nadalol was provided in rat chow (50 g/rat/day). Social defeat (SD) was performed for 7 consecutive days. In SD, the Sprague Dawley rat was introduced into the home cage of a larger, aggressive male Long Evans (LE) rat. The LE rat perceived the Sprague Dawley rat as an intruder, leading to an attack on the rat. The Sprague Dawley rat was forced into a supine postion repeatedly, inducing depression‐like behavior. We used four rat groups; SD group, SD+ Propranolol, SD+ Nadalol, and naïve control. We performed depression‐like behavior tests 24 hours after SD or control exposures, including forced swim test (FST) and social interaction (SIT). In FST, the rat was placed into a cylindrical water‐filled container for five minutes. The total time spent immobile was recorded. Higher immobility time in the water indicates depression‐like behavior. SD rats had significantly more immobility time as compared to control rats in FST. SD rats treated with Propranolol or Nadolol did not exhibit high immobility in FST. SIT measures animal sociability. The Sprage Dawley rats were placed into an apparatus containing two chambers; an empty chamber and a chamber housing a novel rat. SD rats exhibited higher depression‐like behavior (less interaction time with the novel rat as compared to an empty cup) compared to controls. SD rats pretreated with Propranolol or Nadolol did not exhibit low social interaction when compared to SD rats alone. These results suggest that depression‐like behavior induced by SD was prevented when rats were pre‐treated with Propranolol and Nadolol.Support or Funding InformationGrant Support: 2R15MH093918‐02
Combination of Gestational Sub‐Therapeutic Prozac with Tryptophan Depletion Sex Dependently Worsens Social Behavior in Adolescence
Authors: Garbarino Valentina Rose , Edwards Marshall Thomas , Gutierrez Ednia , Santos Taylor , Javors Martin , Daws Lynette C. , Gould Georgianna Grimshaw 
Published: November 2023

Depression is a serious mood disorder that can require antidepressant use during pregnancy. Safety concerns have been raised about the effects of selective‐serotonin reuptake inhibiting (SSRI) antidepressants on fetal brain development. One of the most common SSRIs prescribed during pregnancy is fluoxetine (Prozac). Fluoxetine blocks serotonin (5‐HT) reuptake by the 5‐HT transporter (SERT). 5‐HT is vital for shaping neural circuitry during development, and aberrant fetal 5‐HT levels can increase the risk of autism risk, which is diagnosed with the presence of social behavior deficits and repetitive behaviors. Placental transfer studies show fetal 5‐HT availability may be highly dependent upon maternal availability of its precursor, tryptophan (TRP). Hence, elucidation of the relationship between maternal and fetal TRP and 5‐HT demand and availability is crucial to understanding their role in autism susceptibility. Using a mouse model of maternal SSRI exposure with dietary TRP manipulations during pregnancy, we measured the social interaction and social novelty preference (3 Chamber Test for Social Interaction and Novelty Preference), as well as the repetitive behavior (Marble Burying Test) of adolescent offspring. We hypothesized a daily sub‐therapeutic dose (1 mg/kg/day) of fluoxetine, would impair offspring sociability with relevance to autistic behaviors. In prior studies higher fluoxetine doses increased 5‐HT turnover. Hence we predicted that if a low dose SSRI was combined with dietary TRP deficiency, offspring could develop persistent sociability deficits. Our preliminary findings partially support this hypothesis, as either prenatal fluoxetine exposure or TRP depletion alone, or in combination, impaired sociability preference, in a sex dependent and sociability test phase dependent manner. Treated males seem to lose interaction preference, spending less time with a social interaction stimulus, but maintain normal preference for social novelty based on dwelling time in chambers. Females differentially demonstrate normal social interaction preference, but show a loss of preference for social novelty with fluoxetine treatment in measures of time spent in chamber and time sniffing social or novel stimuli. Additionally, in a measure of repetitive behavior, females exposed to TRP depletion or the combined treatment buried more marbles than males. Our early results suggest that females may be more sensitive to in utero exposure to 5‐HT altering factors, and that males and females may compensate for their developmentally altering effects by different mechanisms.Support or Funding InformationResearch supported by NICHD: HD081261. V. Garbarino is supported by a Biology of Aging Training Grant: T32 AG021890.
Adenylyl Cyclase Isoform Specific Effects in Lipid Raft and Non‐Lipid Raft Membrane Domains Regulate cAMP Compartmentation in Human Airway Smooth Muscle Cells
Authors: Agarwal Shailesh R. , Miyashiro Kathryn , Latt Htun , Ostrom Rennolds S. , Harvey Robert D. 
Published: November 2023

The formation of distinct macromolecular signaling complexes allows different G‐protein coupled receptors to produce diverse functional responses, even while sharing a common second messenger such as cAMP. In human airway smooth muscle (HASM) cells, segregation of specific receptors into different membrane microdomains is thought to critically aid in generating compartmentalized cAMP responses. Whereas, E type prostaglandin receptors (EPRs) have been shown to be expressed in non‐lipid raft domains of the plasma membrane, β‐Adrenergic receptors (βARs) are predominantly expressed in caveolar lipid rafts. In the present study, we tested the hypothesis that adenylyl cyclase type 2 (AC2) preferentially couples to EPRs in a non‐lipid raft domain, while adenylyl cyclase type 6 (AC6) selectively couples to βARs in lipid rafts. To do this, we examined the effect of overexpressing AC2 and AC6 on cAMP responses detected using genetically‐encoded FRET‐based biosensors targeted to lipid raft and non‐lipid raft domains of the plasma membrane, as well as the bulk cytosolic compartment in HASM cells. This approach has the advantage of measuring the kinetics of cAMP production in living cells without the use of PDE inhibitors. Overexpression of AC2 enhanced the cAMP response to EPR activation associated with non‐lipid raft domains, without significantly affecting responses detected elsewhere. AC2 overexpression also had no effect on cAMP responses to βAR activation detected in any subcellular location. These data confirm the hypothesis that AC2 couples exclusively with EPRs in a non‐lipid raft membrane compartment. Overexpression of AC6, on the other hand, actually decreased the response to βAR stimulation associated with lipid rafts, without significantly affecting responses elsewhere. AC6 overexpression also had no effect on the responses to EPR activation detected anywhere in the cell. The ability of AC6 overexpression to inhibit βAR production of cAMP in lipid raft domains was reversed by inhibition of PDE4 activity with rolipram. It was also reversed by overexpression of Ht31 peptide, which disrupts the interaction of protein kinase A with A‐kinase anchoring proteins (AKAPs). These results suggests that AC6 overexpression upregulates and/or recruits PKA and PDE4 activity, which then reduces βAR production of cAMP associated specifically with lipid raft domains.Support or Funding InformationSupport by: NIH R01 GM107094
Region specific proteomic analysis of murine brain after chronic nicotine or menthol
Authors: Mulcahy Matthew J , Wang Jonathan H , Lester Henry A 
Published: November 2023

Nicotinic acetylcholine receptors (nAChRs) are pentameric cation channels that are expressed in the mammalian central nervous system, the peripheral nervous system, and the neuromuscular junction. Eleven neuronal nAChR subunits have been identified in mammals (α2‐7, α9‐10, β2‐4). Chronic administration of nicotine has been shown to increase total α4 and β2 subunit expression in specific brain regions. Menthol, a common cigarette flavoring, has also been shown to upregulate α4 and β2 subunit expression in the cortex, hippocampus, and striatum independent of nicotine co‐administration. These data highlight the potential use of changes in β2 nAChR subunit expression as a marker for neuronal nicotine exposure when administered alone and when administered with additional bioactive compounds such as menthol. We investigated the effect of nicotine and of menthol in several murine brain regions on changes in total β2 subunit protein expression using western blot analysis. Absence of β2 immunoreactivity was confirmed in both β2 knockout and α4 knockout mice. To assess the effects of nicotine, male C57bl/6 mice were separated into two treatment groups: control (saline) and nicotine (2mg/kg/hr). Vehicle (ethanol) and menthol alone (2mg/kg/hr) were compared separately. All treatments were administered for 10–12 days. Nicotine‐induced β2 immunoreactivity was significantly upregulated in isolated brain regions, including the cortex, hippocampus, midbrain, striatum, and thalamus. No change in immunoreactivity was observed in the habenula of mice treated chronically with nicotine. In response to chronic menthol treatment, a significant increase was only observed in the hypothalamus. These data indicate that nicotine and menthol differentially effect expression of nAChR subtypes in various regions of the brain. A comprehensive analysis of changes in β2 nAChR subunit expression in various brain regions will facilitate our understanding of the effects of nicotine exposure in the mammalian CNS. Additionally, the validation of a reliable marker for nicotine and menthol exposure in regions associated with various aspects of addiction and dependence would be invaluable in the pursuit of better cessation therapeutics. Using β2 expression as a marker of nicotine exposure will enable a better understanding of possible effects of nicotine and menthol.Support or Funding InformationThis work is supported by NIH DA036061.
VU590 inhibits Kir1.1 and Kir7.1 potassium channels through distinct molecular mechanisms
Authors: Kharade Sujay V. , Figueroa Eric E. , Denton Jerod S. 
Published: November 2023

Inward rectifier potassium (Kir) channels play key roles in diverse physiological processes and may represent unexploited drug targets for cardiovascular, metabolic, and neurological diseases. However, the lack of potent and selective small‐molecule tool compounds has slowed efforts to explore the integrative physiology and therapeutic value of most Kir channels. In an effort to begin evaluating the druggability of the emerging diuretic target Kir1.1 (ROMK, KCNJ1), we performed a high‐throughput screen of approximately 225,000 compounds for inhibitors of Kir1.1. One compound, termed VU590, was found to inhibit Kir1.1 with an 50% inhibition concentration (IC50) of approximately 0.2 μM, have no appreciable effect on Kir2.1 or Kir4.1, and inhibit Kir7.1 with an IC50 of approximately 8 μM (Lewis et al., 2009). VU590 has since been used to uncover new roles of Kir7.1 in regulation of uterine muscle excitability (McCloskey et al., 2014) and melanocortin signaling in the brain (Ghamari‐Langroudi et al, 2015). Here, we set out to understand the molecular mechanisms by which VU590 inhibits Kir1.1 and Kir7.1. Voltage clamp measurements revealed that block of both Kir1.1 and Kir7.1 is voltage dependent and reduced by increasing the electrochemical driving force for K+, suggesting the VU590 binding site in both channels is located in the pore. Mutation of Asn 171 to Asp (N171D) or Gln (N171Q) in Kir1.1 led to a ~75‐fold and ~4‐fold loss of VU590 potency, respectively. We therefore hypothesized that the negatively charged residue at the equivalent position in Kir7.1 (E149) accounted for the lower potency of VU590 toward Kir7.1. However, the E149Q mutation reduced, whereas the E149D mutation enhanced block of Kir7.1 by VU590. These data suggest that the ligand‐channel interface is fundamentally different between the two channels. By swapping equivalent residues between Kir1.1 and Kir7.1 pore we found that the Kir7.1‐T153C mutation enhanced block of the channel by ~10‐fold, prompting us to examine the mechanism further. Results from comparative homology modeling, mutagenesis, and medicinal chemistry suggest that T153 acts as an energetic barrier to low‐affinity ligands attempting to access a deeper‐pore binding site. This study provides new insights into the molecular pharmacology and structure of Kir1.1 and Kir7.1.Support or Funding InformationSupported by R01DK082884 (JSD)
Anagyrine Desensitization of Peripheral Neuronal Nicotinic Acetylcholine Receptors
Authors: Green Ben , Welch Kevin , Lee Stephen 
Published: November 2023

Members of the genusLupinus cause crooked calf syndrome in the Western United States. Thissyndrome consists of multiple congenital contracture‐type deformities, and isdue to the inhibition of fetal movement by plant alkaloids found in teratogeniclupines. Anagyrine, a teratogenic quinolizidine alkaloid, has been proposed tobe metabolized to a piperidine alkaloid that inhibits fetal movement, theputative mechanism behind crooked calf syndrome. In this research using SH‐SY5Ycells expressing autonomic nicotinic acetylcholine receptors (nAChR) and TE‐671cells expressing fetal muscle‐type nAChR we tested an alternative hypothesisthat anagyrine directly desensitizes fetal muscle‐type nAChR. Unexpectedly, anagyrine was a more potent desensitizer of nAChR expressed by SH‐SY5Y cellswith a fifty percent desensitization concentration (DC50) value of 0.02 μM in SH‐SY5Y cells compared to a DC50 value of 14.7 μM inTE‐671 cells. By comparison, dimethylphenylpiperazinium (DMPP) a nAChR agonistselective for α3β4 nAChR expressed by SHSY‐5Y cells had a DC50 valueof 0.8 μM. In this research, suggeststhat anagyrine is an extremely potent desensitizer of neuronal nAChR which isin contrast to teratogenic piperidine alkaloids.Support or Funding InformationUSDA‐ARS
Discovery and Pharmacological Characterization of Selective Serotonin (5‐HT)2C Receptor Positive Allosteric Modulators (PAMs)
Authors: Wold Eric A. , Wild Christopher T. , McAllister Carrie , Ding Ye , Anastasio Noelle C. , Fox Robert G. , Stutz Sonja , White Mark A. , Chen Haiying , Allen John A. , Cunningham Kathryn A. , Zhou Jia 
Published: November 2023

G protein‐coupled receptors (GPCRs) that are located in the central nervous system (CNS) maintain a fundamental role in modulating healthy and diseased brain states. The serotonin (5‐HT)2C receptor, a CNS predominant GPCR, is a regulatory component of neurobehavioral processes, and decreased signaling at this receptor may contribute to mental health disorders such as impulsivity disorders, drug addiction, schizophrenia and depression. The 5‐HT2C receptor is one of 14 functionally distinct 5‐HT receptors that share topologically similar orthosteric sites, lending importance to subtype selective modulation. Positive allosteric modulation of the 5‐HT2C receptor provides an attractive strategy to selectively fine‐tune binding and/or signaling in response to 5‐HT in a site‐ and event‐specific manner. To this end, we designed, synthesized and pharmacologically characterized a series of 4‐alkylpiperidine‐2‐carboxamides as positive allosteric modulators (PAMs) of the 5‐HT2C receptor. Our efforts have resulted in a series of small molecules that enhance 5‐HT‐evoked Cai2+ release through 5‐HT2C receptor signaling by up to 25% over 5‐HT alone, in vitro. Importantly, no significant effect on 5‐HT‐evoked Cai2+ release through 5‐HT2A receptor signaling was observed. Additionally, these PAMs maintained no intrinsic activity on either the 5‐HT2C receptor or the 5‐HT2A receptor in absence of 5‐HT, suggesting selective 5‐HT2C receptor positive allosteric modulation. Our most efficacious PAM in vitro, CYD‐1‐79, displayed favorable pharmacokinetic parameters and was screened to determine no interaction at all additional 5‐HT receptor orthosteric sites. Excitingly, behavioral studies in rat, using CYD‐1‐79, demonstrated suppression of impulsive action in a dose‐dependent manner by 60% at the highest dose (2 mg/kg), in a one‐choice serial reaction time (1‐CSRT) task, and significantly suppressed context‐induced and cue‐reinforced cocaine seeking (1 mg/kg). These efforts have yielded a selective and efficacious 5‐HT2C receptor PAM that exhibits promising pharmacokinetic properties and modulates 5‐HT2C receptor‐associated behaviors in vivo in a dose‐dependent manner. This novel series of 5‐HT2C receptor PAMs may provide valuable tool compounds to elucidate neuropathological serotonergic function and provides the framework for the development of novel neurotherapeutics.Support or Funding InformationSupported by grants R01 DA038446 (JZ/KAC), K05 DA020087 (KAC), P30 DA028821 (KAC), T32 DA07287 (CTW, EAW), F31 DA038922 (CTW)
TUG‐891, an agonist of GPR120, inhibits lipid accumulation in hepatocytes
Authors: Park Soojin , Kang Saeromi , Im Dongsoon 
Published: November 2023

BackgroundOmega‐3 polyunsaturated fatty acids have protective functions on non‐alcoholic fatty liver disease. Recently, a G protein‐coupled receptor named GPR120 was identified as a receptor for omega‐3 polyunsaturated fatty acids and its protective functions in insulin resistance and hepatic steatosis. Also, GPR120 expression in hepatocytes has also been shown from liver biopsies from child fatty liver disease patients.METHODSIn order to assess functional roles of GPR120 in hepatocytes, an in vitro model of liver X receptor‐mediated hepatocellular steatosis was used.RESULTSGPR120 expression was confirmed in human hepatoma cells. A specific liver X receptor activator induced lipid accumulation and TUG‐891, an agonist of GPR120, inhibited the lipid accumulation. TUG‐891‐induced inhibition was ameliorated by treatment of siRNA transfection of GPR120 or AH7614 (GPR120 antagonist) treatment. SREBP‐1c, a key transcription factor for lipogenesis, was induced by liver X receptor activation. The SREBP‐1c induction was also inhibited by TUG‐891 at mRNA and protein levels. TUG‐891‐induced suppression of SREBP‐1c expression was again blunted by GPR120‐knockdown in the hepatocytes. Therefore, the present data suggest that GPR120 in hepatocytes may function as a receptor for omega‐3 polyunsaturated fatty acids to protect lipid accumulation in the liver.CONCLUSIONProtective functions of omega‐3 polyunsaturated fatty acids on non‐alcoholic fatty liver disease may be mediated by interaction with GPR120 in hepatocytes.
Characterization of an In Vitro Assay for the Detection of Drug‐Induced Cholestasis Using the HepaRG Cell Line
Authors: Krause Caitlin , Blomme Eric , Liguori Michael 
Published: November 2023

Drug‐induced cholestasis (DIC) is a relevant concern in the pharmaceutical industry, and reliable early detection of DIC is of great value in the discovery and development of safe drugs. Traditional animal models are poorly predictive of cholestasis risk in humans, partly because of biological differences. Moreover, limited robust in vitro methods to predict DIC are available. Recently, an in vitro model for the detection of DIC was developed using primary hepatocytes in sandwich configuration. Following incubation of cells with test compounds in the presence or absence of a concentrated bile acid mixture, a DIC Index (DICI) is calculated as a measure of the relative reduction in cellular function. This model was evaluated as part of the European Federation of Pharmaceutical Industries and Associations (EFPIA) Innovative Medicine Initiative project MIP‐DILI. Here, we assessed several test compounds in a modified version of this assay using the HepaRG cell line. Several tool compounds with or without known cholestasis risk were tested at relevant concentrations: five compounds with clinical cholestasis reports (troglitazone, cyclosporin A, chlorpromazine, bosentan, and nefazodone), one toxicant inducing biliary injury (ANIT), two non‐cholestatic compounds known to induce hepatocyte injury (APAP, perhexiline), and one compound associated with rare instances of liver injury in humans (pioglitazone). Albumin and ATP production were assessed as parameters to evaluate the overall functionality and integrity of the cells and DICIs were calculated. Based on change in these parameters relative to the vehicle, the assay accurately predicted 6/6 cholestasis positive compounds and 2/3 cholestasis negative compounds. Results were repeatable. From these data, the HepaRG model for the detection of DIC may be beneficial for hazard identification (i.e., biliary injury) during lead optimization.Support or Funding InformationAuthor Disclosures: All authors are employees of AbbVie. The design, study conduct, and financial support for this research was provided by AbbVie. AbbVie participated in the interpretation of data, review, and approval of the publication.
PMN mediated endothelial cell TRPM2 activation regulates neutrophil trafficking and lung vascular permeability
Authors: Mittal Manish , Nepal Saroj , Tiruppathi Chinnaswamy , Malik Asrar B 
Published: November 2023

TRPM2 (Transient receptor potential melastatin‐2) is a cation channel that mediates Ca2+ entry in response to intracellular ADP ribose (ADPR) generated during oxidative stress. We tested the hypothesis that Reactive Oxygen Species (ROS) released by polymorphonuclear leukocytes (PMNs) during sepsis activate endothelial TRPM2 and induce trans‐endothelial PMN migration and cause lung vascular injury. To investigate the role of endothelial TRPM2 in PMN transmigration, we generated tamoxifen inducible endothelial cell‐restricted TRPM2 knockout (Trpm2ΔEC−/−) mice. Lung vascular permeability and PMN transmigration following LPS challenge (10 mg/Kg) did not increase in these mice. Human lung microvascular endothelial cells (HLMVECs) treated with TRPM2‐siRNA or poly ADP ribose polymerase 1 (PARP1)‐siRNA showed significantly reduced Ca2+ entry in response to activated PMNs (PMN‐to‐EC ratio of 5:1). PMNs isolated from gp91phox−/− mice or WT mice treated with DPI (10 μM) significantly reduced Ca2+ entry in ECs as compared to WT PMNs. ECs treated with PARP1‐siRNA also showed significantly reduced ADPR generation in response to H2O2 stimulation. Overexpression of the ADPR insensitive TRPM2 mutant channel (C1008A) in HLMVECs significantly suppressed the H2O2 induced Ca2+entry. In vivo forced expression of the TRPM2 mutant channel or silencing of PARP‐1 protected against LPS‐induced increase in lung vascular permeability and PMN transmigration in mice. We conclude that endothelial Ca2+ entry following activation of TRPM2 in PMNs disrupts endothelial barrier, thereby promotes lung vascular injury.Support or Funding InformationRO1 HL 077806, RO1 HL 122157