bluuebunny/crossref_metadata_2025_split huggingface dataset:
Macrophage‐Specific Phenotypes Direct Fibroblast Expression of Matrix Metallopeptidases
Authors: Julian Dana Rose , Johnson Zariel , LoPresti Samuel , Brown Bryan , Yates Cecelia 
Published: June 2021

IntroductionDuring wound repair, fibroblasts secrete abundant extracellular matrix (ECM) components and participate in remodeling of the provisional ECM. Macrophages and other immune cells play a critical role in all stages of wound healing and are primarily responsible for directing ECM‐producing cells into the wound site to replace it with the new scar tissue. Classically activated macrophages, M1M□, contribute to inflammation early on, whereas alternatively activated macrophages, M2M□, secrete ECM‐degrading enzymes, including matrix metallopeptidases (MMPs), during resolution. Macrophages instruct fibroblast activities, largely via secreted factors. However, the differential effects of M1M□vs M2M□ on fibroblasts remains unknown. This study is designed to evaluate the various roles of differentially polarized macrophages through secreted factors on fibroblast activities that relate to wound resolution.MethodNaïve macrophages(M0) were differentiated into either M1M□ using IFN‐gamma and LPS, or M2M□ using IL‐4. To study the effects of secreted factors and cell‐contact dependent factors, we used indirect (condition media) and direct macrophages‐fibroblast co‐culture systems, respectively with and without TGFβ. Morphologic, phenotypic, and transcriptomic characterization, and cytokine analysis was performed. Cultures were also evaluated for relative gene expression of extracellular matrix genes (MMP8, MMP9, TIMP1, and TIMP2).ResultsWe have shown that M1M□ increases gene expression of matrix degrading enzymes, MMP8 and MMP9, while M2M□ enhanced fibroblast expression of matrix signaling proteins ERK, p38 and AKT. We further have shown that M1M□ highly expresses chemokine CXCL10, a known inhibitor of fibroblast matrix production. We use CXCR3−/− fibroblasts, as CXCR3 is the only known receptor of CXCL10, and treated the cells with both M1M□ vs M2M□ condition media. The fibroblasts showed no change in MMP8 gene expression. However, when we knocked out CXCR3 in M1M□ and cultured them with fibroblasts, fibroblast MMP8 and MM9 expression was diminished.ConclusionThese data suggest that macrophage‐induction of fibroblast expression of MMP8 and MM9 is independent of CXCR3 expression on fibroblasts, but requires the expression of CXCR3 on macrophages. These data points to complex macrophage‐fibroblast crosstalk coordination and the dependency in tissue repair.Support or Funding InformationFunding Source: American Society of Investigative Pathology‐ Summer Research Opportunity in Pathology Program (SROPP), and University of Pittsburgh School of Nursing supported by grants from NIAMS and NIGMS (AR68317 CCY), and support in kind from and the School of Nursing.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sex Differences in Local Nucleus Accumbens Circuitry and Motivated Behavior
Authors: Brady Lillian J , Johnson Amy R , Tat Jennifer , Calipari Erin S 
Published: June 2021

Drug addiction is a chronic, relapsing brain disease characterized by compulsive drug seeking, high levels of drug intake, and repeated cycles of abstinence and relapse. Historically, preclinical addiction research has focused almost entirely on male subjects; however, in humans, women take and use drugs in larger quantities, transition to addiction faster, and are more likely to relapse, thus representing a particularly vulnerable population. There are a variety of factors that could contribute to these sex differences – including estrous cycle dependent ovarian hormone fluctuations ‐ however the precise neural circuits and neurobiological mechanisms that underlie these differences in reward, motivation, and addiction vulnerability is largely unknown. The mesolimbic dopamine pathway, which connects the ventral tegmental area (VTA) to the nucleus accumbens (NAc), is an essential component of this process as it controls motivation and reward‐seeking behavior. To understand how these phenotypic differences in addiction vulnerability could be driven by estrous cycle‐dependent neurochemical mechanisms we took a multipronged approach to outlining sex‐differences in local circuit regulation of VTA dopamine terminals in the NAc. Using fast‐scan cyclic voltammetry (FSCV) paired with site‐specific pharmacology we measured subsecond dopamine kinetics in male and female mice in either diestrus (low circulating hormones) or estrus (high circulating hormones) and defined sex‐differences in local circuit regulation of dopamine release in the NAc. We found an enhanced dynamic range of calcium and presynaptic D2 autoreceptor control of dopamine release in females compared to males. During estrus, D2 autoreceptors were less effective at inhibiting dopamine release. Further, the relationship between calcium and dopamine release was enhanced in estrus. Finally, cholinergic regulation of dopamine signaling in the NAc was altered in a way that promotes and enhances dopamine release in estrus females. Together, these factors converge to increase the responsivity of dopamine terminals in the NAc to incoming stimuli and likely act to drive enhanced motivation and drug effects on this system. Moving forward it will be critical to directly link these sex‐differences to addiction vulnerability.Support or Funding InformationFunding was provided by startup funds from Vanderbilt University School of Medicine Department of Pharmacology (E.S.C, L.J.B.), as well as funding from the National Institutes of Health (NIH). Funds from the National Institute of Drug Abuse (NIDA) DA042111(E.S.C), National Institute of Mental Health (NIMH) MH065215 (A.R.J), the Brain and Behavior Research Foundation (to E.S.C), Whitehall Foundation (to E.S.C), Edward J Mallinckrodt Jr. Foundation (to E.S.C) also supported this work.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 CF102, an Adenosine A3 Receptors Agonist, on Memory Deficits and Oxidative Stress in a Mouse Model of Streptozotocin‐Induced Dementia of Alzheimer's Type
Authors: Andrade Geanne Matos , Bezerra Jessica Rabelo , de Souza Nascimento Tyciane , Tavares Juliete , Alves Amanda Aragão , Carmo Marta Regina Santos do 
Published: June 2021

Alzheimer's disease (AD) is an age‐related neurodegenerative disease characterized by a progressive decline in cognitive functions. Adenosine receptors play a pivotal role in disease progression, however, adenosine A3 receptors are understudied and their effects vastly unknown. We now investigated the effects of the activation of adenosine A3 receptors on memory deficits and oxidative damage in a mouse model of streptozotocin‐induced experimental dementia of Alzheimer's type. Male Swiss mice received bilateral intracerebroventricular injections of streptozotocin (STZ, 3 mg/kg) dissolved in artificial cerebrospinal fluid. Two days after the first STZ administration, injections were repeated, and the animals were treated with the A3 receptor agonist, CF102 (200 μg/kg intraperitoneally), or vehicle (2% DMSO in saline), 1 h after surgery and once a day until the last day of behavioral evaluation (18th after the first STZ injection). The animals subjected to STZ administration showed significant recognition memory deficits evaluated by the object recognition task, spatial memory deficits evaluated by water maze test, and early and late memory deficits in the passive avoidance test. No differences of locomotor activity in the open field test were observed between groups. STZ also increased malonaldehyde and nitrite contents in the hippocampus. The treatment with CF102 significantly prevented the early and late memory deficits and object recognition memory deficits. CF102 treatment also reduced the increase of nitrite production in 37% in the hippocampus when compared to STZ group. These data highlight the therapeutic potential of A3 agonists in AD, however the role of these receptors on STZ‐induced AD model requires more investigation.Support or Funding InformationSupported by Brazilian National Research and Development Council (CNPq), Coordination for the Improvement of Higher Education Personnel (CAPES) ‐ Brazil, and the Research Support Foundation of Ceará (FUNCAP) – Brazil.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cognition Assessment in APPPS1 Transgenic Mice
Authors: Theobald Robert James , Han Byung Henry 
Published: June 2021

In a previous study, the effects of various anti‐muscarinic drugs were tested on wild type mice using a Barnes Maze to assess cognition. With the availability of the APPPS1 transgenic mouse model of Alzheimer's disease, this study plans to use the Barnes Maze protocol to assess these drug's effects on wild type and transgenic mice in this strain. However, there are no baseline data using the Barnes Maze with this strain, either wild type or transgenic, and the age of onset of cognition deficits in the transgenic mice has not been fully characterized. Therefore, this study was performed to determine if the Barnes Maze was an effective assessment tool in this strain, and if it could be used to determine the age of onset of cognition problems in these mice. Newly born mice were genotyped and both APPPS1 and littermate wild type mice were used in this study. Once the mice reached 3 months of age, they were trained and tested using the Barnes Maze. There were 10 mice tested, 7 wild type (WT) and 3 transgenic mice (APP/PS1 Tg). The mice were tested 3 times per week, Monday, Wednesday, and Friday for approximately 6 months, to establish a pattern of learning the location of an escape box (EsB), and if or when they manifested signs of cognitive failure. Parameters measured included, time to locate the EsB, considered latency, distance traveled to the EsB, and time to enter the EsB. After completion of the behavioral tests, amyloid plaque load in the brain was assessed with methoxy‐ X04 staining. Results indicate that the APPPS1 Tg mice experienced some cognitive failure during the testing period, about 50 days into the testing period. This is manifested as an increase in all three parameters. The WT data shows some variability in performance, but their performance was reasonably consistent after an initial ‘learning’ phase. The data suggests that the Barnes Maze protocol can be used to study the development of cognition failure in the APPPS1 Tg mice, and may be useful in assessing effects of various drugs in these mice.Support or Funding InformationNo extramural fundingThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
microRNA‐195 Alleviates Cognitive Impairment in Rat following Chronic Brain Hypoperfusion
Authors: Zhang Shuai , Xu Yi , Sun Li‐Hua , Chen Xin , Liu Cheng‐Di , Ai Jing 
Published: June 2021

ObjectiveTo observe whether chronic brain hypoperfusion (CBH) induced cognitive decline is associated with amyloid‐β (Aβ)‐cascade response process, dendritic degeneration and neuron death in the hippocampus as well as septal‐hippocampal cholinergic circuit disorder.Methods and resultsWe found that bilateral common carotid artery occlusion (2VO) generated CBH could induce significant cognitive decline and decreased fEPSP of septal‐hippocampal cholinergic circuit in rats, assessed by Morris water maze and in vivo electrophysiological recording. Subsequently, we found the overexpressed protein levels of amyloid precursor protein (APP) and β‐site APP cleaving enzyme 1 (BACE1), death receptor 6 (DR6), together with an activated cdk5/p25, glycogen synthase kinase 3β (GSK3β), inactivated protein phosphatase 2A, degenerated dendrites and neuron death in the hippocampus, as evaluated by western blotting, Golgi, TUNEL and immunofluorescence staining. Interestingly, microRNA‐195 (miR‐195) was found decreased in both hippocampus and basal forebrain. By bioinformatic prediction, we found that miR‐195 could target on 3’‐untranslated regions of Aβ cascade response related genes, including APP, BACE1, Cdk5r1, PME‐1 and Death receptor 6. Further study revealed that miR‐ 195 regulated the expression of APP, BACE1, p35, methylesterase (PME‐1) and DR6 protein at the post‐transcriptional level, as assessed by Dual luciferase reporter assay, siRNA and miR‐masking techniques. In vivo, knockdown of endogenous miR‐195 by lentiviral vector‐mediated over‐expression of its antisense molecule (lenti‐pre‐AMO‐195) elicited the phenotype of dementia and upregulated protein levels of APP, BACE1, p35, PME‐1 and DR6 in rats, whereas over‐expression of miR‐195 using lenti‐pre‐miR‐195 reduced dementia‐vulnerability and those protein levels triggered by CBH. Additionally, chromatin immunoprecipitation analysis showed that NFκB was bound to the promoter region of miR‐195 and inhibited its expression.ConclusionCBH could induce cognitive impairment involving Aβ aggregation, hyperphosphorylation of Tau, GSK3β activation, protein phosphatase‐2A inactivation, degeneration of dendrites and neuron death, which is associated with chronic brain hypoxia‐activated NFκB that inhibited endogenous biogenesis of miR‐195. Down‐regulation of miR‐195 up‐regulated the expression of a battery of Aβ cascade response related proteins at the post‐transcriptional level. MiR‐195 may play a key role in determining dementia susceptibility in CBH rats through a multi‐target way at the post‐transcriptional level, and exogenous complement of miR‐195 may be a potentially valuable anti‐dementia approach.Support or Funding InformationFoundation: This work was supported by Natural Science Foundation of China (81271207, 81070882, 81000499, 81471115, 81671052 and 81371211).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Glutamatergic Antagonism in the Antidepressant‐like Activity of the Rapid‐Acting Antidepressant RO‐25‐6981 and Its Analogs
Authors: Laymon Kristen , Suriyachottakul Jiratchaya , Ham Ahrom , Yu BaoLing , Shim Bora , Valls Danielle C , Kirsh Robert D , So Christopher H , Rawlins David B , Talbot Jeffery N 
Published: June 2021

The glutamatergic antagonist, RO‐25‐6981, has previously been shown to exert both rapid and sustained antidepressant‐like activity. The purpose of the current study is to delineate its putative antidepressant mechanisms of NR2B‐selective NMDA receptor antagonism and monoamine reuptake transporter inhibition. RO‐25‐6981 and structural analogs with varying degrees of NR2B binding affinity were tested for antidepressant‐like behavior in wild‐type mice and animals deficient in NR2B subunit expression. In the tail suspension test, four RO‐25‐6981 analogs (TR‐2, TR‐4, TR‐5, and TR‐6) were found to exhibit antidepressant‐like activity in wild‐type mice following acute administration (30 mg/kg, i.p., 30 min) with maximal reductions in immobility by approximately half compared to vehicle‐treated controls. By contrast, RO‐25‐6981 (10 mg/kg, i.p., 30 min) reduced immobility by approximately 90%, an effect comparable to that exhibited by the traditional monoaminergic antidepressants fluoxetine and desipramine. However, unlike RO‐25‐6981, TR‐2, TR‐4, TR‐5 and TR‐6 profoundly limited generalized locomotor activity suggesting increased activity in the tail suspension test was related to psychotropic vs. generalized drug effects. In contrast, other TR analogs tested showed no antidepressant‐like activity in the tail suspension test, despite possessing robust NMDA receptor antagonist activity via mid‐ to low‐nanomolar binding affinity at the NR2B subunit. Interestingly, RO‐25‐6981 and TR‐5 exhibited similar antidepressant‐like activity in wild‐type and NR2B‐deficient mice, despite possessing low‐nanomolar NR2B binding. In addition, cellular serotonin transport assays showed functional inhibitory activity of both agents. Taken together, these data suggest that the antidepressant‐like activity of RO‐25‐6981 and its analogs does not correlate with the degree of NMDA receptor antagonism. Furthermore, these data point to monoamine reuptake inhibition contributing to the overall antidepressant‐like activity of RO‐25‐6981 in animal models of mood.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protein kinase C‐beta isoform as a mechanism promoting chronic pain in sickle cell disease
Authors: Wang Zaijie Jim , He Ying 
Published: June 2021

Pain is a hallmark symptom in sickle cell disease (SCD). Besides acute painful vaso‐occlusive crises, SCD is also accompanied by intractable chronic pain. This persistent, and often unrelieved, pain starts early in childhood and continues throughout life. The neurobiological mechanisms of chronic pain in SCD remain unclear, which limits effective pain management and the quality of life in patients with SCD. Taking advantage of a humanized mouse model of SCD, this study aimed to investigate protein kinase C (PKC)‐beta isoform mechanism for chronic pain in SCD. We characterized pain phenotypes in the Townes' mouse model (TOW) of SCD that exclusively expresses human alleles encoding normal α‐ and sickle β‐globin. TOW mice exhibited ongoing spontaneous pain as well as hypersensitivity to evoked pain stimuli compared with littermate control mice. We then examined spinal PKCbeta‐mediated signaling and behavioral consequences. Prominent activation of PKCbeta was identified in the superficial laminae of the spinal cord in TOW sickle mice. Functional inhibition and genetic silencing of the PKCbeta attenuated sickle cell pain phenotypes, abolishing PKCbeta activation, spontaneous pain, mechanical allodynia, and heat hyperalgesia in the sickle cell mice. Taken together, these data suggest PKCbeta may be one of the mechanisms for the generation and maintenance of ongoing and evoked pain in SCD. These findings offer new insights into sickle cell pain mechanisms, which may offer new pharmacological interventions.Support or Funding InformationR35 HL140031 and K99 HL133590This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Vesicular Glutamate Transporter 2 in Neuropathic Pain and Its Regulation Mechanism
Authors: Su Rui‐Bin , Yu Gang 
Published: June 2021

BACKGROUNDUp‐regulation of glutamate neurotransmission has been proposed as a crucial mechanism of neuropathic pain. Vesicular glutamate transporters (VGLUTs), which transport glutamate into presynaptic vesicles, are fundamental in glutamate storage and release. Several lines of evidence from genetically modified mice suggest a possible association of VGLUT2 with neuropathic pain.OBJECTIVEThe purpose of this study is to investigate the changes of VGLUT2 expression and function in neuropathic pain and to elucidate its regulatory mechanism, in a mouse spared nerve injury (SNI) model.METHODSThe changes in VGLUT2 expression were determined by western blot and immunohistochemistry in the spinal cord and pain‐related brain regions at different time points after SNI surgery. The cell distribution characteristics of VGLUT2 in the pathological condition were observed by immunofluorescent staining. RNA interference and VGLUT inhibitors were used to regulate the expression and function of VGLUT2 respectively, and the mechanical pain behavior was observed. The effects of VGLUT2 down‐regulation and inhibition on glutamate release were also investigated. The effects of Wnt/β‐catenin signaling on VGLUT2 expression and pain behavior were investigated by using the Wnt agonist, Wnt1, and Wnt/β‐catenin pathway inhibitor XAV939 in SNI mice.RESULTSVGLUT2 protein levels were significantly up‐regulated in the spinal cord, thalamus, periaqueductal gray and amygdala after SNI surgery. In SNI mice, VGLUT2 was mainly expressed in neurons (co‐locolized with NeuN and MAP2), but not glia cells (Iba‐1 and GFAP). Intrathecal or intracerebral administration of VGLUT2 shRNAs or the VGLUT inhibitor CSB6B significantly attenuated mechanical allodynia and decreased glutamate release. Meanwhile, Wnt1/β‐catenin signaling pathway was activated during the development of neuropathic pain. The Wnt agonist or Wnt1 significantly increased VGLUT2 expression, while Wnt/β‐catenin pathway inhibitor XAV939 decreased VGLUT2 expression. Moreover, administration of XAV939 7 days after SNI surgery significantly attenuated mechanical allodynia with a long duration of action, and this effect was in coincidence with VGLUT2 down‐regulation. In naïve mice, intrathecal administration of the Wnt agonist or Wnt1 produced mechanical allodynia, which was abolished by preventive VGLUT2 down‐regulation.CONCLUSIONNeuropathic pain are associated with the changes in VGLUT2 expression and function, and Wnt1/β‐catenin signaling up‐regulates VGLUT2 in the pathological condition.Support or Funding InformationThis work was supported by National Natural Science Foundation of China (81200850) and Beijing Natural Science Foundation (7123224).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Calmodulin interaction with GRK5 N‐terminus regulates kinase activation
Authors: Sulon Sarah Marie , Komolov Konstantin , Bhardwaj Anshul , van Keulen Siri Camee , Dror Ron , Benovic Jeffrey 
Published: June 2021

GRK5 belongs to a family of G protein‐coupled receptor (GPCR) kinases that mediate phosphorylation and desensitization of GPCRs. In addition, GRK5 has been implicated in a number of diseases through its ability to phosphorylate non‐receptor substrates such as α‐synuclein, HDAC5 and moesin. GRK5 is directly regulated by the calcium‐sensing protein calmodulin (CaM). Ca2+/CaM binds GRK5 and inhibits GPCR phosphorylation, though the precise mechanism of regulation is unclear. Interestingly, GRK5 retains kinase activity when in complex with CaM. To better understand the structural basis of GRK5 regulation by CaM, we utilized crystallography, molecular dynamics simulations, isothermal titration calorimetry, mutagenesis and in vitro kinase assays. The 2.0 Å crystal structure of the GRK5/CaM complex reveals that the far N‐terminal α‐helical region (αN‐helix) of GRK5 interacts with the CaM N‐lobe via a hydrophobic interface. The CaM‐bound structure of GRK5 exhibits key conformational changes that are characteristic of kinase activation. These include disruption of the ionic lock, ordering of the αN‐helix and closure of the catalytic domain. In addition, CaM stabilizes GRK5 in an open, active conformation and stimulates non‐receptor substrate phosphorylation. The stabilization of the αN‐helix by CaM likely serves as the mechanism of activation. Thus, CaM binding to GRK5 via a novel N‐terminal binding site stabilizes an active conformation of the kinase, enhancing phosphorylation of non‐receptor substrates.Support or Funding InformationROI HL142310This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.