bluuebunny/crossref_metadata_2025_split huggingface dataset:
Oxycodone‐induced gene expression adaptations in the brain reward center in a murine model of neuropathic pain
Authors: Pryce Kerri D. , Kronman Hope , Ramakrishnan Aarthi , Gaspari Sevasti , Pena Catherine , Shen Li , Nestler Eric , Zachariou Venetia 
Published: June 2021

Chronic neuropathic pain is a complex condition with complications beyond physical symptoms that include depression, anxiety, and cognitive issues. One in five patients suffering from chronic pain is prescribed an opiate drug such as oxycodone for pain relief. Long‐term use of oxycodone and other opiates for the treatment of chronic pain can lead to physical dependence and addiction. Recent studies from our group revealed that chronic neuropathic pain promotes several adaptations in the brain reward regions responsible for both the sensory and affective components associated with chronic pain (Descalzi et al., 2017, Sci. Signaling). Here, we are using Next Generation RNA‐sequencing to gain insight on the gene expression adaptations triggered by persistent exposure to oxycodone and to determine the impact of neuropathic pain in maladaptive plasticity to oxycodone. We employed the spared nerve injury model (SNI) of neuropathic pain in adult C57Bl/6 mice and assessed peripheral neuropathy symptoms (thermal hyperalgesia, mechanical allodynia) for ~9 weeks. Mice were then treated daily with oxycodone, for two weeks and gene expression was monitored three weeks after oxycodone withdrawal. Oxycodone treatment induced thermal hyperalgesia regardless of pain states, while withdrawal produced significant mechanical allodynia. We next monitored changes in gene expression for three brain regions implicated in the symptoms of chronic neuropathic pain: medial prefrontal cortex (mPFC), nucleus accumbens (NAc) and the ventral tegmental area (VTA). Although oxycodone treatment promotes mostly unique transcriptome profile across brain regions, we observed similar downstream effectors and transcription factors to be affected in pain states when compared with non‐pain states. We are currently validating these genes using transgenic mouse lines to investigate their roles in potentiating oxycodone induced analgesia as well as physical dependence and addiction‐like behaviors. Understanding the gene adaptions in the brain after oxycodone treatment provides novel insight into the cellular machinery involved in maladaptive responses to drugs of abuse in the presence and absence of pain.Support or Funding Information5T32DA007135‐34, NIDA, R01 NS086444‐01, NINDS, and P01 DA008227, NIDAThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Combinations of haloperidol and morphine produce maximal levels of anti‐hyperalgesic or potentiation effects in rats with neuropathic pain
Authors: MENA‐VALDES LICET CARIDAD , ESPINOSA‐JUAREZ JOSUE VIDAL , BLANCO‐HERNANDEZ YISEL , LOPEZ‐MUÑOZ FRANCISCO JAVIER 
Published: June 2021

Neuropathic pain (NP) is a chronic disorder that result from an injury or dysfunction at central or peripheral nervous system, promoting alterations in control and modulation of nociceptive signalling pathways. The haloperidol is a butyrophenone prototype, that recently some authors have described as anti‐nociceptive drug mediated by sigma‐1 receptors antagonism. Besides, morphine is a prototype analgesic opioid drug currently used in the NP treatment.ObjectiveTo detect the optimal combinations generated by therapy of haloperidol with morphine in NP induced by chronic constriction injury (CCI).MethodsThe anti‐hyperalgesic effects of haloperidol (0.017–0.100 mg/Kg, s.c.) and morphine (0.01–3.16 mg/Kg, s.c.) were determined after single‐doses, both in monotherapy and combined, using the von Frey test. Evaluations were done until 10 days post‐surgery at 30, 60, 90, 120 and 180 minutes after drugs administration and data were processed using Surface of Synergic Interaction analysis (SSI).ResultsHaloperidol and morphine showed a dose‐dependent anti‐hyperalgesic effects. From 16 combinations studied, six were potentiation, while remaining 10 combinations resulted in an additive effect or limited by the maximum effect of the individual doses. The combination morphine (0.0316 mg/Kg) and haloperidol (0.01778 mg/Kg) showed the maximum potentiation effect (p<0.001). In turn, the combination of maximum anti‐hyperalgesic efficacy (p<0.01) was morphine (1.0 mg/Kg) and haloperidol (0.1 mg/Kg). Interestingly, was detected that is required a haloperidol single dose tenfold higher than haloperidol dose employed by the combination of maximum potentiation for reach similar antinociceptive effect.ConclusionsThis study showed that haloperidol and morphine in combination can produce addition and potentiation effects in neuropathic pain, and the SSI analysis allowed to detect the optimal combinations to produce maximum efficacy or maximum potentiation of anti‐hyperalgesic effects.Support or Funding InformationDepartment of Pharmacobiology, Cinvestav‐Unidad Coapa, C.P. 14330, Tlálpan, Mexico City, Mexico.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chronic and Intermittent Ethanol Treatment Differentially Regulates Membrane Compartmentalization of G‐protein Subunits in Rat Brain
Authors: Miller James , Wiley Aidan , Luessen Deborah , McCool Brian , Chen Rong 
Published: June 2021

IntroductionEthanol exposure produces profound changes in the trafficking and activity of many G‐protein coupled receptors (GPCRs) which are associated with alcohol preference, tolerance, and dependence. However, the molecular mechanism involved in ethanol‐induced changes in receptor function remains unclear. Our lab previously found that ethanol treatment increases cholesterol content in rat brain, altering membrane lipid composition. Cholesterol, enriched in lipid rafts of membranes, plays an important role in membrane integrity and promotes the recruitment of signaling molecules. Ethanol‐induced disruption of brain cholesterol homeostasis would influence the signaling of many membrane‐bound GPCRs and channels. This study aimed to examine whether CIE also causes changes in the membrane localization of G‐protein subunits which are tightly associated with many GPCRs and channels.MethodsMale Sprague Dawley rats were exposed to 7 cycles of 12 hours of ethanol vapor inhalation followed by 12 hours of normal air. Rats were sacrificed 24 hours after the final ethanol exposure and the prefrontal cortex (PFC) tissue was dissected. Samples were homogenized followed by discontinuous sucrose density (5%–40%) ultracentrifugation and separated by density into 15 fractions. Western blotting was performed to determine the localization of G‐protein subunits on in lipid raft and non‐lipid raft fractions.ResultsCIE selectively altered membrane compartmentalization of certain subunits of G‐protein. There was an increase in localization of Gαo subunit from lipid rafts to non‐lipid rafts in CIE‐treated rat PFC. In contrast, CIE caused Gβ subunit to translocate from non‐lipid rafts to lipid rafts. Moreover, CIE had no effect on membrane compartmentalization of Gαi3 and Gαq. These results indicate selective changes in location of certain G‐protein subunits, altering proximal microdomain interaction with membrane channels and receptors and thus downstream signaling. This provides a possible mechanism underlying cholesterol‐mediated changes in GPCR signaling in rat brain tissue.Support or Funding InformationThis project is supported by NIH R01DA042862, P50DA006634, F31AA025532.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phospholipase Cɛ Regulation of Cardiac Fibroblasts
Authors: Brown Loren M. , Rodriguez Soneida S. , Smrcka Alan V. 
Published: June 2021

Heart disease is one of the leading causes of death in the United States. Cardiac fibrosis is commonly associated with heart diseases, such as cardiac hypertrophy and myocardial infarctions. This is characterized as an excess deposition of extracellular matrix that results in a stiffening of the heart. Cardiac fibrosis can lead to arrhythmias, decreased diastolic function and eventually heart failure. Upon injury to the heart, cardiac fibroblasts transition to an activated myofibroblast state, characterized by the expression of α‐smooth muscle actin (αSMA). While the activation of fibroblasts is initially beneficial to the heart, chronic activation of fibroblasts leads to excessive deposition of collagen resulting in the development of fibrosis. Phospholipase Cɛ (PLCɛ) is an important signaling enzyme that is activated by small GTPases downstream of G‐protein coupled receptors (GPCRs) and receptor tyrosine kinases and is important in the cardiac myocyte for the development of cardiac hypertrophy. In this study we sought to elucidate the role of PLCɛ in the transition of cardiac fibroblasts to activated myofibroblasts. Using neonatal rat cardiac fibroblasts, we demonstrate that PLCɛ is highly expressed in fibroblasts compared to myocytes at the mRNA and protein level. Adenovirus encoding for a PLCɛ‐shRNA was used to knockdown PLCɛ in fibroblasts and the ability of the cells to transition to a myofibroblast was examined. Knocking down PLCɛ decreases the mRNA expression of pro‐fibrotic genes such as αSMA, collagen 1, and TGFβ indicating that PLCɛ can regulate pro‐fibrotic gene expression in cardiac fibroblasts. The transition to myofibroblasts induced by transforming growth factor‐β (TGFβ) is reduced in cells treated with PLCɛ‐shRNA compared to cells treated with a control shRNA as assessed by staining of αSMA. In addition, knocking down PLCɛ reduces the ability of the cardiac fibroblasts to respond to pro‐fibrotic Gq‐coupled GPCRs such as the angiotensin receptor and the protease‐activated receptor 1. These data indicate that PLCɛ has an important role in pro‐fibrotic signaling in cardiac fibroblasts and can regulate the myofibroblast transition. Future studies will focus on elucidating the molecular mechanism by which PLCɛ is involved in the pro‐fibrotic signaling response.Support or Funding InformationThis project is supported by grant number T32‐HL007853 from the NIHThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regulation of the Ca2+‐sensing capacity of STIM1 by the G protein‐coupled estrogen receptor
Authors: Pekas Devon , Gebert‐Oberle Briana , Giles Jennifer , Underwood Timothy , VerMeer Mark , Tran Kim 
Published: June 2021

The stromal interaction molecule 1 (STIM1) is essential for activation of store‐operated Ca2+entry (SOCE). STIM1 contains in its intra‐endoplasmic reticular (ER) domain a canonical EF hand (cEF) that houses a Ca2+‐binding loop. This loop serves as a Ca2+sensor; Ca2+leaving its binding loop upon ER Ca2+depletion initiates conformational changes leading to STIM1 oligomerization and SOCE activation. We recently reported that activation of the G protein‐coupled estrogen receptor (GPER) clamps intracellular Ca2+signals via multifaceted inhibitory actions on both Ca2+efflux and SOCE; the latter results partly from alteration in phosphorylation at serines 575, 608 and 621 of STIM1. Here, we identified a dynamic interaction between GPER and STIM1 that directly controls the Ca2+‐sensing capacity of STIM1. GPER coimmunoprecipitates with STIM1 in resting primary endothelial cells, an association enhanced by treatment with either GPER agonist G‐1 or activation of SOCE using thapsigargin. This effect is not altered by combined S575/608/621A substitutions. Coimmunoprecipitation analysis using truncated versions of STIM1 showed that GPER associates with the cEF domain. Confocal microscopy identified a GPER's submembrane domain that localizes GPER to the ER. To assess the role of GPER‐STIM1 interaction in controlling STIM1's Ca2+sensing capacity, we generated a series of biosensors that incorporated STIM1's Ca2+‐binding loop in singlet, doublet or triplet. These biosensors faithfully report Ca2+binding to the loop, with tripled dynamic range by the triple‐loop biosensor and loss of Ca2+binding by charge substitutions targeting strategic elements for Ca2+binding. Using peptides corresponding to submembrane domains of GPER and these STIM1 biosensors, we identified a robust Ca2+‐independent interaction between a submembrane domain and the Ca2+‐binding loop. The charge substitutions that abolish Ca2+binding only reduce but do not negate the interaction, indicating that Ca2+and the GPER domain bind to different regions of the loop. Consistently, we have observed that GPER overexpression decreases SOCE and GPER knockdown substantially increases SOCE. These data reveal a new interaction between GPER and STIM1 that directly controls STIM1's Ca2+‐sensing capacity and SOCE activation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CCK1 ‐ a Very Peculiar G Protein‐Coupled Receptor
Authors: Cui Zong Jie 
Published: June 2021

The class A G protein‐coupled receptor cholecystokinin 1 (CCK1) plays a predominant role in regulating exocrine pancreatic secretion both in mammals and in birds. CCK1R supra‐maximal stimulation leads to one of the most commonly used murine models of acute pancreatitis. The CCK‐CCK1R system is a well conserved feeding regulator in evolution from worms, insects, other invertebrates, vertebrates, to primates. In the murine brain, CCK1R is mainly expressed in the hippocampus and a few highly defined extracortical regions. These expression patterns determine that CCK1R modulation is important in both biology and medicine. Indeed some special characteristics revealed recently have made CCK1 stand out among all the class A GPCR. Although CCK1 is not susceptible to superoxide or hydrogen peroxide oxidative regulation, it is permanently activated by singlet oxygen (1O2) which can be generated conveniently in a Type II photodynamic action. We have used Fura‐2 fluorescent calcium imaging to show that CCK1R is activated permanently by photodynamic action with photosensitiser sulphonated aluminium phthalocyanine (SALPC) in the freshly isolated rat pancreatic acini. Such permanent photodynamic activation of CCK1R with SALPC could be reproduced in rat pancreatic acinar tumor cell AR4‐2J, and in cell lines ectopically expressing human hCCK1R. Both CCK1R in AR4‐2J and ectopically expressed in other cell lines (CHO, HEK) could be permanently activated by photodynamic action with the genetically encoded protein photosensitisers KillerRed and miniSOG. Other than permanent activation by 1O2, CCK1R activation by CCK in the freshly isolated rat pancreatic acini is quenched by mixed reactive oxygen species generated by neutrophil respiratory burst, and by neutrophil extracellular trap component histones. Further, CCK1R activation by CCK or by 1O2 in AR4‐2J is sensitized by histones. Our ongoing work is examining the potential for CCK1R analogues from invertebrates to be permanently activated by photodynamic action with SALPC, KillerRed, miniSOG or their variants. We are also looking at the possibility to extend the permanent activation by 1O2 (pAb1O2) property of CCK1R to other GPCR by the generation of chimeric receptors. The outcomes of such experiments will provide useful toolkits for remote photon‐driven activations of the CCK1R system in vivo from invertebrates to vertebrates, birds, mammals and non‐human primates.Support or Funding InformationThis work was supported by The Natural Science Foundation of China (31670856).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Synthesis, Characterization and Evaluation of Cadmium Sulfide Nanoparticles Capped with Dextrin and Functionalized with Temozolomide
Authors: Leo‐Campos Frida , Rodriguez Patricia , Reyes‐Esparza Jorge , Rodriguez‐Fragoso Lourdes 
Published: June 2021

Nanoparticles are currently used as drug carriers. Temozolomide is an antineoplastic drug for tumor in central nervous system. It presents a half‐life of 1.8 hours. For this reason, requires high doses that are limited by side effects. Studies about the use of nanoparticles as drugs transporters have reported an increase the drug solubility, increase in half‐life and sustained release of the drug because it is protected from degradation and loss of therapeutic effect. This group has development and studied CdS nanoparticles capped with polymers and have showed being biocompatible and effective as drug carriers. To synthesize and characterize cadmium sulfide nanoparticles capped with dextrin functionalized with temozolomide and observe cell uptake. By means of the chemical method, the precursors were allowed to react to obtain the synthesis of CdS/dextrin nanoparticles. Subsequently, the nanoparticles were functionalized with temozolomide through a chemical method in which the OH groups of the dextrin were bounded to the NH2 Through the technique of Fourier‐transform infrared, the characteristic bonds were observed: at 840 cm−1 the α‐D glucopyranose bond, at 927 cm−1 the α(1–4) bonds; 1159 cm−1 glucopyranosyl units and at 1079 cm−1 the C‐O‐C, at 1025 cm−1 the C‐O bond, at 3399 cm−1 the OH group characteristic of dextrin. At 2001 cm−1 the C=N=N of diazomethane was found which is a toxic compound resulting from the formation of nanoparticles. Also at 3400 cm−1 there is the link N=H and 1610 cm−1 C = C or C = N of the temozolomide. According to the X‐ray results, the widening of 3 peaks corresponding to the crystalline planes characteristic of the cubic phase of the CdS is observed which are: (111), (220) and (311). Using the Scherrer formula d = 0.8λ/βcos θ was calculated that the average size of the nanoparticles is 3 nm. Due to the size of the semiconductor nanoparticles of CdS‐dextrin, they emit green fluorescence characteristic. Therefore, tumor cells treated with nanoparticles functionalized with temozolomide in epifluorescence microscope were observed and it was observed that the nanoparticles reached the cytoplasm and core of cells and produces the effect of temozolomide arresting cell cycle in phase G2 + M. The synthesis and characterization of CdS/dextrin nanoparticles functionalized with temozolomide resulted in 5nm, which are uptake by the cells and are distributed on the cytoplasm and core exerting the effect of the drug. The CdS/dextrin has the potential for being used as drug carriers and facilitating the pharmacological because cells up take them.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Sub‐Chronic Inhalation of Crude Oil Vapor (COV) on Reactivity to Methacholine and Neural Regulation of Rat Airways in vitro
Authors: Fedan Jeffrey S , Russ Kristen A , Thompson Janet A , Jackson Mark C , McKinney Walter G 
Published: June 2021

Inhalation of crude oil vapor (COV), a mixture of hydrocarbon vapors and volatile organic compounds (VOCs), by oil and gas industry workers has led to a number of deaths. Workers are potentially exposed to COV during sampling, gauging and filling crude oil storage tanks. Inasmuch as the effects of COV inhalation on various functions of the lung is undescribed, we have been investigating the effects of COV inhalation in a rat model. Earlier, we found increases in transepithelial resistance 1 d post‐exposure to COV (300ppm total VOCs, 6 h/d×1 d; whole body chambers). Lung resistance was increased 90 dpost‐exposure, but no changes occurred in lung compliance, airway reactivity in vivo, or epithelial ion transport in vitro, 1, 28 and 90 d after this acute exposure. Here, rats were exposed to COV sub‐chronically (300 ppm total VOCs, 6 h/d×28 d), and airway reactivity tomethacholine (MCh) in vitro and neurogenic responses of airway smooth muscle in vitro were investigated 1, 28 and 90 d post‐exposure. Reactivity to MCh and the modulatory role of the airway epithelium was assessed by comparing contractile responses of the isolated, perfused trachea to extraluminally (EL)‐and intraluminally (IL)‐applied MCh. Reactivity to EL‐applied MCh was unaffected, but reactivity to IL‐applied MCh was decreased at 90 d post‐exposure, indicative of an enhanced inhibitory effect of the epithelium. To investigate the effect of sub‐chronic COV exposure on neural regulation of airway smooth muscle, tracheal strips were placed in an organchamber between two platinum ring electrodes to record isometric tension. Electric field stimulation (EFS) was delivered across the electrodes to stimulate frequency‐dependent contractile responses of the muscle. Exposure of rats to COV had no effect on EFS‐evoked contractions at any post‐exposure time point, indicating that neurons releasing acetylcholine in the airway wall were not a target of COV. The results indicate that the airway epithelium is affected by inhalation of COV, and that cholinergic motor nerves are not. Physiological systems in the lung have different, exposure time‐ and post‐exposure time‐dependent susceptibilities to inhaled COV's effects.Support or Funding InformationNIOSH 7939ZYACThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Oncogenic Gαq and Gα11 Inhibition by FR900359 in Uveal Melanoma
Authors: Lapadula Dominic , Farias Eduardo , Randolph Clinita E. , Purwin Timothy J. , McGrath Dougan , Charpentier Thomas H. , Zhang Lihong , Wu Shihua , Terai Mizue , Sato Takami , Tall Gregory G. , Zhou Naiming , Wedegaertner Philip B. , Aplin Andrew E. , Aguirre‐Ghiso Julio , Benovic Jeffrey L. 
Published: June 2021

Uveal melanoma is the most common intraocular tumor in adults and often metastasizes to the liver, leaving patients with few options. Recurrent activating mutations in the G proteins, Gαq and Gα11, are observed in approximately 93% of all uveal melanomas. While therapeutic intervention of downstream Gαq/11 targets has been unsuccessful in treating uveal melanoma, using in vitro [35S]GTPγS binding and cell signaling assays we have found that the Gαq/11 inhibitor, FR900359 (FR), effectively inhibits oncogenic Gαq/11 signaling in uveal melanoma cells expressing either mutant Gαq or Gα11. Inhibition of oncogenic Gαq/11 by FR results in cell cycle arrest and induction of apoptosis. Furthermore, colony formation is prevented by FR treatment of uveal melanoma cells in 3D‐cell culture, providing promise for future in vivo studies. This suggests direct inhibition of activating Gαq/11 mutants may be a potential means of treating uveal melanoma.Support or Funding InformationDr. Ralph and Marian Falk Medical Research Trust and NIH award F31 CA225064This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.