150 results on '"Prado VF"'
Search Results
2. Influence of β 2 -adrenergic selective agonist formoterol on the motor unit of a mouse model of a congenital myasthenic syndrome with complete VAChT deletion.
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Rossi L, Mota BI, Valadão PAC, Magalhães-Gomes MPS, Oliveira BS, Guatimosim S, Navegantes LCC, Miranda AS, Prado MAM, Prado VF, and Guatimosim C
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Congenital Myasthenic Syndromes (CMS) are a set of genetic diseases that affect the neuromuscular transmission causing muscular weakness. The standard pharmacological treatment aims at ameliorating the myasthenic symptom by acetylcholinesterase inhibitors. Most patients respond well in the short and medium term, however, over time the beneficial effects rapidly fade, and the efficacy of the treatment diminishes. Increasing evidence shows that β
2 -adrenergic agonists can be a suitable choice for the treatment of neuromuscular disorders, including CMS, as they promote beneficial effects in the neuromuscular system. The exact mechanism on which they rely is not completely understood, although patients and animal models respond well to the treatment, especially over extended periods. Here, we report the use of the long-lasting specific β2 -adrenergic agonist formoterol in a myasthenic mouse model (mnVAChT-KD), featuring deletion of VAChT (Vesicular Acetylcholine Transporter) specifically in the α-motoneurons. Our findings demonstrate that formoterol treatment (300 μg/kg/day; sc) for 30 days increased the neuromuscular junction area, induced skeletal muscle hypertrophy and altered fibre type composition in myasthenic mice. Interestingly, β2 -adrenergic agonists have shown efficacy even in the absence of ACh (acetylcholine). Our data provide important evidence supporting the potential of β2 -adrenergic agonists in treating neuromuscular disorders of pre-synaptic origin and characterized by disruptions in nerve-muscle communication, through a direct and beneficial action within the motor unit., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)- Published
- 2024
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3. Neuroanatomical and cognitive biomarkers of alpha-synuclein propagation in a mouse model of synucleinopathy prior to onset of motor symptoms.
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Tullo S, Miranda AS, Del Cid-Pellitero E, Lim MP, Gallino D, Attaran A, Patel R, Novikov V, Park M, Beraldo FH, Luo W, Shlaifer I, Durcan TM, Bussey TJ, Saksida LM, Fon EA, Prado VF, Prado MAM, and Chakravarty MM
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- Animals, Mice, Humans, Magnetic Resonance Imaging, Male, Female, Cognition physiology, Mice, Transgenic, Reversal Learning physiology, alpha-Synuclein metabolism, Synucleinopathies pathology, Synucleinopathies metabolism, Disease Models, Animal, Biomarkers metabolism
- Abstract
Significant evidence suggests that misfolded alpha-synuclein (aSyn), a major component of Lewy bodies, propagates in a prion-like manner contributing to disease progression in Parkinson's disease (PD) and other synucleinopathies. In fact, timed inoculation of M83 hemizygous mice with recombinant human aSyn preformed fibrils (PFF) has shown symptomatic deficits after substantial spreading of pathogenic alpha-synuclein, as detected by markers for the phosphorylation of S129 of aSyn. However, whether accumulated toxicity impact human-relevant cognitive and structural neuroanatomical measures is not fully understood. Here we performed a single unilateral striatal PFF injection in M83 hemizygous mice, and using two assays with translational potential, ex vivo magnetic resonance imaging (MRI) and touchscreen testing, we examined the combined neuroanatomical and behavioral impact of aSyn propagation. In PFF-injected mice, we observed widespread atrophy in bilateral regions that project to or receive input from the injection site using MRI. We also identified early deficits in reversal learning prior to the emergence of motor symptoms. Our findings highlight a network of regions with related cellular correlates of pathology that follow the progression of aSyn spreading, and that affect brain areas relevant for reversal learning. Our experiments suggest that M83 hemizygous mice injected with human PFF provides a model to understand how misfolded aSyn affects human-relevant pre-clinical measures and suggest that these pre-clinical biomarkers could be used to detect early toxicity of aSyn and provide better translational measures between mice and human disease., (© 2023 The Authors. Journal of Neurochemistry published by John Wiley & Sons Ltd on behalf of International Society for Neurochemistry.)
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- 2024
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4. Alpha-7 Nicotinic Receptor Agonist Protects Mice Against Pulmonary Emphysema Induced by Elastase.
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Banzato R, Pinheiro-Menegasso NM, Novelli FPRS, Olivo CR, Taguchi L, de Oliveira Santos S, Fukuzaki S, Teodoro WPR, Lopes FDTQS, Tibério IFLC, de Toledo-Arruda AC, Prado MAM, Prado VF, and Prado CM
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- Animals, Mice, Male, Lung pathology, Lung drug effects, Lung metabolism, Anti-Inflammatory Agents pharmacology, Anti-Inflammatory Agents therapeutic use, alpha7 Nicotinic Acetylcholine Receptor agonists, alpha7 Nicotinic Acetylcholine Receptor metabolism, Pancreatic Elastase, Pulmonary Emphysema drug therapy, Pulmonary Emphysema chemically induced, Pulmonary Emphysema metabolism, Pulmonary Emphysema prevention & control, Benzamides pharmacology, Benzamides therapeutic use, Bridged Bicyclo Compounds pharmacology, Bridged Bicyclo Compounds therapeutic use, Nicotinic Agonists pharmacology, Nicotinic Agonists therapeutic use, Mice, Inbred C57BL
- Abstract
Pulmonary emphysema is a primary component of chronic obstructive pulmonary disease (COPD), a life-threatening disorder characterized by lung inflammation and restricted airflow, primarily resulting from the destruction of small airways and alveolar walls. Cumulative evidence suggests that nicotinic receptors, especially the α7 subtype (α7nAChR), is required for anti-inflammatory cholinergic responses. We postulated that the stimulation of α7nAChR could offer therapeutic benefits in the context of pulmonary emphysema. To investigate this, we assessed the potential protective effects of PNU-282987, a selective α7nAChR agonist, using an experimental emphysema model. Male mice (C57BL/6) were submitted to a nasal instillation of porcine pancreatic elastase (PPE) (50 µl, 0.667 IU) to induce emphysema. Treatment with PNU-282987 (2.0 mg/kg, ip) was performed pre and post-emphysema induction by measuring anti-inflammatory effects (inflammatory cells, cytokines) as well as anti-remodeling and anti-oxidant effects. Elastase-induced emphysema led to an increase in the number of α7nAChR-positive cells in the lungs. Notably, both groups treated with PNU-282987 (prior to and following emphysema induction) exhibited a significant decrease in the number of α7nAChR-positive cells. Furthermore, both groups treated with PNU-282987 demonstrated decreased levels of macrophages, IL-6, IL-1β, collagen, and elastic fiber deposition. Additionally, both groups exhibited reduced STAT3 phosphorylation and lower levels of SOCS3. Of particular note, in the post-treated group, PNU-282987 successfully attenuated alveolar enlargement, decreased IL-17 and TNF-α levels, and reduced the recruitment of polymorphonuclear cells to the lung parenchyma. Significantly, it is worth noting that MLA, an antagonist of α7nAChR, counteracted the protective effects of PNU-282987 in relation to certain crucial inflammatory parameters. In summary, these findings unequivocally demonstrate the protective abilities of α7nAChR against elastase-induced emphysema, strongly supporting α7nAChR as a pivotal therapeutic target for ameliorating pulmonary emphysema., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
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- 2024
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5. Multimodal gradients of basal forebrain connectivity across the neocortex.
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Chakraborty S, Haast RAM, Onuska KM, Kanel P, Prado MAM, Prado VF, Khan AR, and Schmitz TW
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The cholinergic innervation of the cortex originates almost entirely from populations of neurons in the basal forebrain (BF). Structurally, the ascending BF cholinergic projections are highly branched, with individual cells targeting multiple different cortical regions. However, it is not known whether the structural organization of basal forebrain projections reflects their functional integration with the cortex. We therefore used high-resolution 7T diffusion and resting state functional MRI in humans to examine multimodal gradients of BF cholinergic connectivity with the cortex. Moving from anteromedial to posterolateral BF, we observed reduced tethering between structural and functional connectivity gradients, with the most pronounced dissimilarity localized in the nucleus basalis of Meynert (NbM). The cortical expression of this structure-function gradient revealed progressively weaker tethering moving from unimodal to transmodal cortex, with the lowest tethering in midcingulo-insular cortex. We used human [
18 F] fluoroethoxy-benzovesamicol (FEOBV) PET to demonstrate that cortical areas with higher concentrations of cholinergic innervation tend to exhibit lower tethering between BF structural and functional connectivity, suggesting a pattern of increasingly diffuse axonal arborization. Anterograde viral tracing of cholinergic projections and [18 F] FEOBV PET in mice confirmed a gradient of axonal arborization across individual BF cholinergic neurons. Like humans, cholinergic neurons with the highest arborization project to cingulo-insular areas of the mouse isocortex. Altogether, our findings reveal that BF cholinergic neurons vary in their branch complexity, with certain subpopulations exhibiting greater modularity and others greater diffusivity in the functional integration of their cortical targets.- Published
- 2024
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6. Sex-dependent cholinergic effects on amyloid pathology: A translational study.
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German-Castelan L, Shanks HRC, Gros R, Saito T, Saido TC, Saksida LM, Bussey TJ, Prado MAM, Schmitz TW, and Prado VF
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- Mice, Humans, Female, Male, Animals, Aged, Amyloid beta-Peptides, Estradiol, Cholinergic Agents, Amyloid beta-Protein Precursor, Mice, Transgenic, Disease Models, Animal, Alzheimer Disease pathology, Cognitive Dysfunction
- Abstract
Introduction: About two-thirds of Alzheimer's Disease (AD) patients are women, who exhibit more severe pathology and cognitive decline than men. Whether biological sex causally modulates the relationship between cholinergic signaling and amyloid pathology remains unknown., Methods: We quantified amyloid beta (Aβ) in male and female App-mutant mice with either decreased or increased cholinergic tone and examined the impact of ovariectomy and estradiol replacement in this relationship. We also investigated longitudinal changes in basal forebrain (cholinergic function) and Aβ in elderly individuals., Results: We show a causal relationship between cholinergic tone and amyloid pathology in males and ovariectomized female mice, which is decoupled in ovary-intact and ovariectomized females receiving estradiol. In elderly humans, cholinergic loss exacerbates Aβ., Discussion: Our findings emphasize the importance of reflecting human menopause in mouse models. They also support a role for therapies targeting estradiol and cholinergic signaling to reduce Aβ., Highlights: Cholinergic tone regulates amyloid beta (Aβ) pathology in males and ovariectomized female mice. Estradiol uncouples the relationship between cholinergic tone and Aβ. In elderly humans, cholinergic loss correlates with increased Aβ in both sexes., (© 2023 The Authors. Alzheimer's & Dementia published by Wiley Periodicals LLC on behalf of Alzheimer's Association.)
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- 2024
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7. Open science and data sharing in cognitive neuroscience with MouseBytes and MouseBytes.
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Memar S, Jiang E, Prado VF, Saksida LM, Bussey TJ, and Prado MAM
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- Animals, Genomics, Information Dissemination, Magnetic Resonance Imaging, Neuroimaging, Humans, Cognitive Neuroscience, Databases, Factual
- Abstract
Open access to rodent cognitive data has lagged behind the rapid generation of large open-access datasets in other areas of neuroscience, such as neuroimaging and genomics. One contributing factor has been the absence of uniform standardization in experiments and data output, an issue that has particularly plagued studies in animal models. Touchscreen-automated cognitive testing of animal models allows standardized outputs that are compatible with open-access sharing. Touchscreen datasets can be combined with different neuro-technologies such as fiber photometry, miniscopes, optogenetics, and MRI to evaluate the relationship between neural activity and behavior. Here we describe a platform that allows deposition of these data into an open-access repository. This platform, called MouseBytes, is a web-based repository that enables researchers to store, share, visualize, and analyze cognitive data. Here we present the architecture, structure, and the essential infrastructure behind MouseBytes. In addition, we describe MouseBytes+, a database that allows data from complementary neuro-technologies such as imaging and photometry to be easily integrated with behavioral data in MouseBytes to support multi-modal behavioral analysis., (© 2023. The Author(s).)
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- 2023
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8. AAV-mediated neuronal expression of an scFv antibody selective for Aβ oligomers protects synapses and rescues memory in Alzheimer models.
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Selles MC, Fortuna JTS, Cercato MC, Santos LE, Domett L, Bitencourt ALB, Carraro MF, Souza AS, Janickova H, Azevedo CV, Campos HC, de Souza JM, Alves-Leon S, Prado VF, Prado MAM, Epstein AL, Salvetti A, Longo BM, Arancio O, Klein WL, Sebollela A, De Felice FG, Jerusalinsky DA, and Ferreira ST
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- Mice, Rats, Humans, Animals, Aged, Amyloid beta-Peptides genetics, Amyloid beta-Peptides metabolism, Synapses metabolism, Neurons metabolism, Memory Disorders genetics, Memory Disorders therapy, Alzheimer Disease genetics, Alzheimer Disease therapy, Alzheimer Disease metabolism, Single-Chain Antibodies genetics, Single-Chain Antibodies metabolism
- Abstract
The accumulation of soluble oligomers of the amyloid-β peptide (AβOs) in the brain has been implicated in synapse failure and memory impairment in Alzheimer's disease. Here, we initially show that treatment with NUsc1, a single-chain variable-fragment antibody (scFv) that selectively targets a subpopulation of AβOs and shows minimal reactivity to Aβ monomers and fibrils, prevents the inhibition of long-term potentiation in hippocampal slices and memory impairment induced by AβOs in mice. As a therapeutic approach for intracerebral antibody delivery, we developed an adeno-associated virus vector to drive neuronal expression of NUsc1 (AAV-NUsc1) within the brain. Transduction by AAV-NUsc1 induced NUsc1 expression and secretion in adult human brain slices and inhibited AβO binding to neurons and AβO-induced loss of dendritic spines in primary rat hippocampal cultures. Treatment of mice with AAV-NUsc1 prevented memory impairment induced by AβOs and, remarkably, reversed memory deficits in aged APPswe/PS1ΔE9 Alzheimer's disease model mice. These results support the feasibility of immunotherapy using viral vector-mediated gene delivery of NUsc1 or other AβO-specific single-chain antibodies as a potential therapeutic approach in Alzheimer's disease., Competing Interests: Declaration of interests A patent application covering the use of AAV-NUsc1 in Alzheimer’s disease has been filed with the USPTO (16/820,269; pending) by Northwestern University with S.T.F., W.L.K., D.A.J., A. Sebollela, and M.C.S. as named inventors., (Copyright © 2022 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)
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- 2023
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9. Continuous cholinergic-dopaminergic updating in the nucleus accumbens underlies approaches to reward-predicting cues.
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Skirzewski M, Princz-Lebel O, German-Castelan L, Crooks AM, Kim GK, Tarnow SH, Reichelt A, Memar S, Palmer D, Li Y, Jane Rylett R, Saksida LM, Prado VF, Prado MAM, and Bussey TJ
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- Mice, Animals, Acetylcholine, Cues, Cholinergic Agents, Reward, Dopamine, Nucleus Accumbens physiology
- Abstract
The ability to learn Pavlovian associations from environmental cues predicting positive outcomes is critical for survival, motivating adaptive behaviours. This cued-motivated behaviour depends on the nucleus accumbens (NAc). NAc output activity mediated by spiny projecting neurons (SPNs) is regulated by dopamine, but also by cholinergic interneurons (CINs), which can release acetylcholine and glutamate via the activity of the vesicular acetylcholine transporter (VAChT) or the vesicular glutamate transporter (VGLUT3), respectively. Here we investigated behavioural and neurochemical changes in mice performing a touchscreen Pavlovian approach task by recording dopamine, acetylcholine, and calcium dynamics from D1- and D2-SPNs using fibre photometry in control, VAChT or VGLUT3 mutant mice to understand how these signals cooperate in the service of approach behaviours toward reward-predicting cues. We reveal that NAc acetylcholine-dopaminergic signalling is continuously updated to regulate striatal output underlying the acquisition of Pavlovian approach learning toward reward-predicting cues., (© 2022. The Author(s).)
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- 2022
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10. Chemogenetic activation of VGLUT3-expressing neurons decreases movement.
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Kljakic O, Hogan-Cann AE, Yang H, Dover B, Al-Onaizi M, Prado MAM, and Prado VF
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- Neurons, Glutamic Acid, Acetylcholine, Clozapine pharmacology
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Vesicular glutamate transporters (VGLUTs) are responsible for the storage of glutamate into secretory vesicles. The VGLUT3 isoform is mainly expressed in neurons that secrete other classical neurotransmitters, including the cholinergic interneurons in the striatum, and VGLUT3-expressing neurons often secrete two distinct neurotransmitters. VGLUT3 is discretely distributed throughout the brain and is found in subpopulations of spinal cord interneurons, in subset of neurons in the dorsal root ganglion, and in Merkel cells. Mice with a global loss of VGLUT3 are hyperactive and the modulation of specific VGLUT3-expressing circuits can lead to changes in movement. In this study, we tested the hypothesis that increased activity of VGLUT3-expressing neurons is associated with decreased movement. Using a mouse line expressing excitatory designer receptor exclusively activated by designer drugs (hM3Dq-DREADD) on VGLUT3-expressing neurons, we showed that activation of hM3Dq signalling acutely decreased locomotor activity. This decreased locomotion was likely not due to circuit changes mediated by glutamate nor acetylcholine released from VGLUT3-expressing neurons, as activation of hM3Dq signalling in mice that do not release glutamate or acetylcholine from VGLUT3-expressing neurons also decreased locomotor activity. This suggests that other neurotransmitters are likely driving this hypoactive phenotype. We used these mouse lines to compare the effects of DREADD agonists in vivo. We observed that clozapine-N-oxide (CNO), clozapine, compound 21 and perlapine show small differences in the speed at which they prompt behavioural responses but the four of them are selective DREADD ligands., (Copyright © 2022 Elsevier B.V. All rights reserved.)
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- 2022
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11. Stress-inducible phosphoprotein 1 (HOP/STI1/STIP1) regulates the accumulation and toxicity of α-synuclein in vivo.
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Lackie RE, de Miranda AS, Lim MP, Novikov V, Madrer N, Karunatilleke NC, Rutledge BS, Tullo S, Brickenden A, Maitland MER, Greenberg D, Gallino D, Luo W, Attaran A, Shlaifer I, Del Cid Pellitero E, Schild-Poulter C, Durcan TM, Fon EA, Duennwald M, Beraldo FH, Chakravarty MM, Bussey TJ, Saksida LM, Soreq H, Choy WY, Prado VF, and Prado MAM
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- Animals, HSP90 Heat-Shock Proteins chemistry, HSP90 Heat-Shock Proteins metabolism, Humans, Mice, Molecular Chaperones metabolism, Phosphoproteins, Ubiquitins, alpha-Synuclein toxicity, Heat-Shock Proteins metabolism, Intrinsically Disordered Proteins, alpha-Synuclein metabolism
- Abstract
The predominantly pre-synaptic intrinsically disordered protein α-synuclein is prone to misfolding and aggregation in synucleinopathies, such as Parkinson's disease (PD) and Dementia with Lewy bodies (DLB). Molecular chaperones play important roles in protein misfolding diseases and members of the chaperone machinery are often deposited in Lewy bodies. Here, we show that the Hsp90 co-chaperone STI1 co-immunoprecipitated α-synuclein, and co-deposited with Hsp90 and Hsp70 in insoluble protein fractions in two mouse models of α-synuclein misfolding. STI1 and Hsp90 also co-localized extensively with filamentous S129 phosphorylated α-synuclein in ubiquitin-positive inclusions. In PD human brains, STI1 transcripts were increased, and in neurologically healthy brains, STI1 and α-synuclein transcripts correlated. Nuclear Magnetic Resonance (NMR) analyses revealed direct interaction of α-synuclein with STI1 and indicated that the STI1 TPR2A, but not TPR1 or TPR2B domains, interacted with the C-terminal domain of α-synuclein. In vitro, the STI1 TPR2A domain facilitated S129 phosphorylation by Polo-like kinase 3. Moreover, mice over-expressing STI1 and Hsp90ß presented elevated α-synuclein S129 phosphorylation accompanied by inclusions when injected with α-synuclein pre-formed fibrils. In contrast, reduced STI1 function decreased protein inclusion formation, S129 α-synuclein phosphorylation, while mitigating motor and cognitive deficits as well as mesoscopic brain atrophy in α-synuclein-over-expressing mice. Our findings reveal a vicious cycle in which STI1 facilitates the generation and accumulation of toxic α-synuclein conformers, while α-synuclein-induced proteostatic stress increased insoluble STI1 and Hsp90., (© 2022. The Author(s).)
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- 2022
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12. Measuring Nonapoptotic Caspase Activity with a Transgenic Reporter in Mice.
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Nicholls PJ, Pack TF, Urs NM, Kumar S, Zhou Y, Ichim G, Ginzel JD, Turu G, Calabrese E, Roberts WL, Fan P, Ostapchenko VG, Guzman Lenis MS, Beraldo F, Hatina J, Prado VF, Prado MAM, Spasojevic I, Snyder JC, Dzirasa K, Johnson GA, and Caron MG
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- Male, Female, Mice, Animals, Mice, Transgenic, Neuronal Plasticity, Caspase 9, Apoptosis physiology, Brain
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The protease caspase-3 is a key mediator of apoptotic programmed cell death. But weak or transient caspase activity can contribute to neuronal differentiation, axonal pathfinding, and synaptic long-term depression. Despite the importance of sublethal, or nonapoptotic, caspase activity in neurodevelopment and neural plasticity, there has been no simple method for mapping and quantifying nonapoptotic caspase activity (NACA) in rodent brains. We therefore generated a transgenic mouse expressing a highly sensitive and specific fluorescent reporter of caspase activity, with peak signal localized to the nucleus. As a proof of concept, we first obtained evidence that NACA influences neurophysiology in an amygdalar circuit. Then focusing on the amygdala, we were able to quantify a sex-specific persistent elevation in caspase activity in females after restraint stress. This simple in vivo caspase activity reporter will facilitate systems-level studies of apoptotic and nonapoptotic phenomena in behavioral and pathologic models., (Copyright © 2022 Nicholls et al.)
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- 2022
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13. Muscarinic and N-methyl-D-aspartate receptor blockade reveal differences in hippocampal local field potentials in mice with low cholinergic tone.
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Leung LS, Moallem S, Prado MAM, Prado VF, and Chu L
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- Animals, Hippocampus physiology, Mice, Mice, Knockout, Muscarinic Antagonists pharmacology, N-Methylaspartate, Scopolamine pharmacology, Vesicular Acetylcholine Transport Proteins, Acetylcholine, Receptors, N-Methyl-D-Aspartate
- Abstract
We hypothesize that hippocampal local field potentials in acetylcholine (ACh)-deficient mutant mice, compared to wild-type (WT) mice, will show lower sensitivity to muscarinic cholinergic antagonist scopolamine (5 mg/kg i.p.) but higher sensitivity to NMDA receptor antagonist 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP, 10 mg/kg i.p.). Recordings were made during walk and awake-immobility (IMM) in WT mice, and in mice with forebrain knockout (KO) of the vesicular acetylcholine transporter (VAChT) gene, or heterozygous knockdown of VAChT gene (KD). Scopolamine or CPP did not significantly alter walk theta frequency, which was higher in KD than WT/KO mice. Scopolamine decreased theta power peak rise during walk in WT/KD mice but not in KO mice, while CPP suppressed theta peak rise more in WT/KO mice than KD mice. During IMM, scopolamine decreased gamma1 (γ1, 30-58 Hz) power more in KD/WT mice than KO mice, while delta (1-4 Hz) power and delta-gamma cross-frequency coherence (CFC) were increased in all mouse groups during IMM or walk. During walk, scopolamine increased delta and beta (13-30 Hz) power and decreased gamma2 (γ2, 62-100 Hz) power and theta-γ2 CFC more in WT/KD than KO mice. Theta-γ2, but not theta-γ1, CFC increased with theta-peak-frequency in WT/KD mice, and was suppressed by scopolamine at high theta (8-10 Hz) frequency; theta-γ2 CFC in KO mice was not significantly altered by scopolamine. CPP decreased beta and gamma power more in KD/KO mice compared to WT mice, while delta power and delta-gamma CFC were increased in all mouse groups. ACh deficiency exacerbates the attenuation of beta and gamma power by CPP. We conclude that both muscarinic and NMDA transmission contribute toward hippocampal theta, beta, and gamma power, and a decrease in gamma power or theta-gamma CFC may be associated with loss of arousal and cognitive functions., (© 2022 Wiley Periodicals LLC.)
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- 2022
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14. Aerobic exercise training engages cholinergic signaling to improve emphysema induced by cigarette smoke exposure in mice.
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Suehiro CL, Souza NTS, da Silva EB, Cruz MM, Laia RM, de Oliveira Santos S, Santana-Novelli FPR, de Castro TBP, Lopes FDTQS, Pinheiro NM, de Fátima Lopes Calvo Tibério I, Olivo CR, Alonso-Vale MI, Prado MAM, Prado VF, de Toledo-Arruda AC, and Prado CM
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- Animals, Cholinergic Agents, Inflammation, Lung, Mice, Mice, Inbred C57BL, Vesicular Acetylcholine Transport Proteins, Cigarette Smoking, Emphysema, Pulmonary Emphysema etiology, Pulmonary Emphysema prevention & control
- Abstract
Lung inflammation is modulated by cholinergic signaling and exercise training protects mice against pulmonary emphysema development; however, whether exercise training engages cholinergic signaling is unknown., Aims: As cholinergic signaling is directly linked to the vesicular acetylcholine transporter (VAChT) levels, we evaluated whether the effects of aerobic exercise training depend on the VAChT levels in mice with pulmonary emphysema., Main Methods: Wild-type (WT) and mutant (KD
HOM ) mice (65-70% of reduction in VAChT levels) were exposed to cigarette smoke (30 min, 2×/day, 5×/week, 12 weeks) and submitted or not to aerobic exercise training on a treadmill (60 min/day, 5×/week, 12 weeks). Lung function and inflammation were evaluated., Key Findings: Cigarette smoke reduced body mass in mice (p < 0.001) and increased alveolar diameter (p < 0.001), inflammation (p < 0.001) and collagen deposition (p < 0.01) in lung tissue. Both trained groups improved their performance in the final physical test compared to the initial test (p < 0.001). In WT mice, exercise training protected against emphysema development (p < 0.05), reduced mononuclear cells infiltrate (p < 0.001) and increased MAC-2 positive cells in lung parenchyma (p < 0.05); however, these effects were not observed in KDHOM mice. The exercise training reduced iNOS-positive cells (p < 0.001) and collagen fibers deposition (p < 0.05) in lung parenchyma of WT and KDHOM mice, although KDHOM mice showed higher levels of iNOS-positive cells., Significance: Our data suggest that the protective effects of aerobic exercise training on pulmonary emphysema are, at least in part, dependent on the integrity of the lung cholinergic signaling., (Copyright © 2022 Elsevier Inc. All rights reserved.)- Published
- 2022
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15. Neuronal cholinergic signaling constrains norepinephrine activity in the heart.
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Guimarães DA, Aquino NSS, Rocha-Resende C, Jesus ICG, Silva MM, Scalzo SA, Fonseca RC, Durand MT, Pereira V, Tezini GCSV, Oliveira A, Prado VF, Stefanon I, Salgado HC, Prado MAM, Szawka RE, and Guatimosim S
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- Adrenergic Agents, Animals, Mice, Myocytes, Cardiac, Vesicular Acetylcholine Transport Proteins genetics, Cholinergic Agents, Norepinephrine
- Abstract
It is well known that cholinergic hypofunction contributes to cardiac pathology, yet, the mechanisms involved remain unclear. Our previous study has shown that genetically engineered model of cholinergic deficit, the vesicular acetylcholine transporter knockdown homozygous (VAChT KD
HOM ) mice, exhibit pathological cardiac remodeling and a gradual increase in cardiac mass with aging. Given that an increase in cardiac mass is often caused by adrenergic hyperactivity, we hypothesized that VAChT KDHOM mice might have an increase in cardiac norepinephrine (NE) levels. We thus investigated the temporal changes in NE content in the heart from 3-, 6-, and 12-mo-old VAChT mutants. Interestingly, mice with cholinergic hypofunction showed a gradual elevation in cardiac NE content, which was already increased at 6 mo of age. Consistent with this finding, 6-mo-old VAChT KDHOM mice showed enhanced sympathetic activity and a greater abundance of tyrosine hydroxylase positive sympathetic nerves in the heart. VAChT mutants exhibited an increase in peak calcium transient, and mitochondrial oxidative stress in cardiomyocytes along with enhanced G protein-coupled receptor kinase 5 (GRK5) and nuclear factor of activated T-cells (NFAT) staining in the heart. These are known targets of adrenergic signaling in the cell. Moreover, vagotomized-mice displayed an increase in cardiac NE content confirming the data obtained in VAChT KDHOM mice. Establishing a causal relationship between acetylcholine and NE, VAChT KDHOM mice treated with pyridostigmine, a cholinesterase inhibitor, showed reduced cardiac NE content, rescuing the phenotype. Our findings unveil a yet unrecognized role of cholinergic signaling as a modulator of cardiac NE, providing novel insights into the mechanisms that drive autonomic imbalance.- Published
- 2022
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16. Functional dissociation of behavioral effects from acetylcholine and glutamate released from cholinergic striatal interneurons.
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Kljakic O, Janíčková H, Skirzewski M, Reichelt A, Memar S, El Mestikawy S, Li Y, Saksida LM, Bussey TJ, Prado VF, and Prado MAM
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- Animals, Dopamine metabolism, Male, Mice, Mice, Inbred C57BL, Neurotransmitter Agents metabolism, Signal Transduction physiology, Vesicular Acetylcholine Transport Proteins metabolism, Vesicular Glutamate Transport Proteins metabolism, Acetylcholine metabolism, Cholinergic Agents metabolism, Corpus Striatum metabolism, Glutamic Acid metabolism, Interneurons metabolism
- Abstract
In the striatum, cholinergic interneurons (CINs) have the ability to release both acetylcholine and glutamate, due to the expression of the vesicular acetylcholine transporter (VAChT) and the vesicular glutamate transporter 3 (VGLUT3). However, the relationship these neurotransmitters have in the regulation of behavior is not fully understood. Here we used reward-based touchscreen tests in mice to assess the individual and combined contributions of acetylcholine/glutamate co-transmission in behavior. We found that reduced levels of the VAChT from CINs negatively impacted dopamine signalling in response to reward, and disrupted complex responses in a sequential chain of events. In contrast, diminished VGLUT3 levels had somewhat opposite effects. When mutant mice were treated with haloperidol in a cue-based task, the drug did not affect the performance of VAChT mutant mice, whereas VGLUT3 mutant mice were highly sensitive to haloperidol. In mice where both vesicular transporters were deleted from CINs, we observed altered reward-evoked dopaminergic signalling and behavioral deficits that resemble, but were worse, than those in mice with specific loss of VAChT alone. These results demonstrate that the ability to secrete two different neurotransmitters allows CINs to exert complex modulation of a wide range of behaviors., (© 2022 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.)
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- 2022
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17. Evaluating Sequential Response Learning in the Rodent Operant Touchscreen System.
- Author
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Janickova H, Kljakic O, Robbins TW, Saksida LM, Bussey TJ, Prado VF, and Prado MAM
- Subjects
- Animals, Corpus Striatum, Mice, Learning, Rodentia
- Abstract
Sequential and cue-directed response learning in rodents have been previously shown to depend on intact striatal signaling. In particular, these behaviors rely on striatal dopamine and acetylcholine release, with an impairment of sequential response learning evident in animal models with alterations in the two systems. Here we provide a protocol for testing sequential response/response chain learning using the rodent touchscreen system. Specifically, the present protocol is designed to implement the heterogeneous sequence task, adapted from Keeler et al. (2014), in the rodent touchscreen apparatus. This task has been used previously to assess complex motor learning and response selection in mice. In the following protocol, the task is performed in touchscreen-based automated chambers with five response locations using food reinforcers to maintain performance. The sequence task requires the subject to make five nose pokes to white square stimuli appearing in five different locations sequentially from left to right. © 2021 Wiley Periodicals LLC. Basic Protocol: Implementation of the heterogeneous sequence task Support Protocol: Creation of the heterogeneous sequence task ABET II touchscreen schedule., (© 2021 Wiley Periodicals LLC.)
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- 2021
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18. Motoneuron-specific loss of VAChT mimics neuromuscular defects seen in congenital myasthenic syndrome.
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Joviano-Santos JV, Kljakic O, Magalhães-Gomes MPS, Valadão PAC, de Oliveira LR, Prado MAM, Prado VF, and Guatimosim C
- Subjects
- Acetylcholine metabolism, Animals, Disease Models, Animal, Humans, Mice, Motor Neurons pathology, Muscle Contraction genetics, Muscle Contraction physiology, Muscle, Skeletal metabolism, Muscle, Skeletal physiology, Myasthenic Syndromes, Congenital metabolism, Myasthenic Syndromes, Congenital pathology, Neuromuscular Junction genetics, Neuromuscular Junction metabolism, Neurotransmitter Agents genetics, Spinal Cord metabolism, Spinal Cord physiology, Synaptic Transmission genetics, Synaptic Vesicles metabolism, Acetylcholine genetics, Motor Neurons metabolism, Myasthenic Syndromes, Congenital genetics, Vesicular Acetylcholine Transport Proteins genetics
- Abstract
Motoneurons (MNs) control muscle activity by releasing the neurotransmitter acetylcholine (ACh) at the level of neuromuscular junctions. ACh is packaged into synaptic vesicles by the vesicular ACh transporter (VAChT), and disruptions in its release can impair muscle contraction, as seen in congenital myasthenic syndromes (CMS). Recently, VAChT gene mutations were identified in humans displaying varying degrees of myasthenia. Moreover, mice with a global deficiency in VAChT expression display several characteristics of CMS. Despite these findings, little is known about how a long-term decrease in VAChT expression in vivo affects MNs structure and function. Using Cre-loxP technology, we generated a mouse model where VAChT is deleted in select groups of MNs (mnVAChT-KD). Molecular analysis revealed that the VAChT deletion was specific to MNs and affected approximately 50% of its population in the brainstem and spinal cord, with alpha-MNs primarily targeted (70% in spinal cord). Within each animal, the cell body area of VAChT-deleted MNs was significantly smaller compared to MNs with VAChT preserved. Likewise, muscles innervated by VAChT-deleted MNs showed atrophy while muscles innervated by VAChT-containing neurons appeared normal. In addition, mnVAChT KD mice had decreased muscle strength, were hypoactive, leaner and exhibited kyphosis. This neuromuscular dysfunction was evident at 2 months of age and became progressively worse by 6 months. Treatment of mutants with a cholinesterase inhibitor was able to improve some of the motor deficits. As these observations mimic what is seen in CMS, this new line could be valuable for assessing the efficacy of potential CMS drugs., (© 2021 Federation of European Biochemical Societies.)
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- 2021
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19. Cholinergic transmission from the basal forebrain modulates social memory in male mice.
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Kljakic O, Al-Onaizi M, Janíčková H, Chen KS, Guzman MS, Prado MAM, and Prado VF
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- Acetylcholine, Animals, Cholinergic Agents, Hippocampus metabolism, Male, Mice, Vesicular Acetylcholine Transport Proteins metabolism, Basal Forebrain
- Abstract
Disruptions in social behaviour are prevalent in many neuropsychiatric disorders such as schizophrenia, bipolar disorder and autism spectrum disorders. However, the underlying neurochemical regulation of social behaviour is still not well understood. The central cholinergic system has been proposed to contribute to the regulation of social behaviour. For instance, decreased global levels of acetylcholine release in the brain leads to decreased social interaction and an impairment of social memory in mice. Nonetheless, it has been difficult to ascertain the specific brain areas where cholinergic signalling influences social preference and social memory. In this study, we investigated the impact of different forebrain cholinergic regions on social behaviour by examining mouse lines that differ in their regional expression level of the vesicular acetylcholine transporter-the protein that regulates acetylcholine secretion. We found that when cholinergic signalling is highly disrupted in the striatum, hippocampus, cortex and amygdala mice have intact social preference but are impaired in social memory, as they cannot remember a familiar conspecific nor recognize a novel one. A similar pattern emerges when acetylcholine release is disrupted mainly in the striatum, cortex, and amygdala; however, the ability to recognize novel conspecifics is retained. In contrast, cholinergic signalling of the striatum and amygdala does not appear to significantly contribute to the modulation of social memory and social preference. Furthermore, we demonstrated that increasing global cholinergic tone does not increase social behaviours. Together, these data suggest that cholinergic transmission from the hippocampus and cortex are important for regulating social memory., (© 2021 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.)
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- 2021
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20. Long-term endogenous acetylcholine deficiency potentiates pulmonary inflammation in a murine model of elastase-induced emphysema.
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Banzato R, Pinheiro NM, Olivo CR, Santana FR, Lopes FDTQS, Caperuto LC, Câmara NO, Martins MA, Tibério IFLC, Prado MAM, Prado VF, and Prado CM
- Subjects
- Acetylcholine metabolism, Animals, Bronchoalveolar Lavage Fluid cytology, Cytokines metabolism, Disease Models, Animal, Emphysema metabolism, Inflammation pathology, Lung pathology, Macrophages metabolism, Male, Mice, NF-kappa B metabolism, Neutrophils metabolism, Pancreatic Elastase adverse effects, Pancreatic Elastase pharmacology, Pneumonia physiopathology, Pulmonary Emphysema metabolism, Signal Transduction, Vesicular Acetylcholine Transport Proteins deficiency, Vesicular Acetylcholine Transport Proteins genetics, Vesicular Acetylcholine Transport Proteins metabolism, Acetylcholine deficiency, Emphysema immunology, Pneumonia etiology
- Abstract
Acetylcholine (ACh), the neurotransmitter of the cholinergic system, regulates inflammation in several diseases including pulmonary diseases. ACh is also involved in a non-neuronal mechanism that modulates the innate immune response. Because inflammation and release of pro-inflammatory cytokines are involved in pulmonary emphysema, we hypothesized that vesicular acetylcholine transport protein (VAChT) deficiency, which leads to reduction in ACh release, can modulate lung inflammation in an experimental model of emphysema. Mice with genetical reduced expression of VAChT (VAChT KD
HOM 70%) and wild-type mice (WT) received nasal instillation of 50 uL of porcine pancreatic elastase (PPE) or saline on day 0. Twenty-eight days after, animals were evaluated. Elastase instilled VAChT KDHOM mice presented an increase in macrophages, lymphocytes, and neutrophils in bronchoalveolar lavage fluid and MAC2-positive macrophages in lung tissue and peribronchovascular area that was comparable to that observed in WT mice. Conversely, elastase instilled VAChT KDHOM mice showed significantly larger number of NF-κB-positive cells and isoprostane staining in the peribronchovascular area when compared to elastase-instilled WT-mice. Moreover, elastase-instilled VAChT-deficient mice showed increased MCP-1 levels in the lungs. Other cytokines, extracellular matrix remodeling, alveolar enlargement, and lung function were not worse in elastase-instilled VAChT deficiency than in elastase-instilled WT-controls. These data suggest that decreased VAChT expression may contribute to the pathogenesis of emphysema, at least in part, through NF-κB activation, MCP-1, and oxidative stress pathways. This study highlights novel pathways involved in lung inflammation that may contribute to the development of chronic obstrutive lung disease (COPD) in cholinergic deficient individuals such as Alzheimer's disease patients., (© 2021. The Author(s).)- Published
- 2021
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21. Lung Edema and Mortality Induced by Intestinal Ischemia and Reperfusion Is Regulated by VAChT Levels in Female Mice.
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Santana FPR, Ricardo-da-Silva FY, Fantozzi ET, Pinheiro NM, Tibério IFLC, Moreira LFP, Prado MAM, Prado VF, Tavares-de-Lima W, Prado CM, and Breithaupt-Faloppa AC
- Subjects
- Animals, Female, Inflammation Mediators metabolism, Intestines blood supply, Mice, Mice, Transgenic, Ovariectomy adverse effects, Ovariectomy mortality, Intestines metabolism, Pulmonary Edema metabolism, Pulmonary Edema mortality, Reperfusion Injury metabolism, Reperfusion Injury mortality, Vesicular Acetylcholine Transport Proteins metabolism
- Abstract
Acute lung injury induced by intestinal ischemia/reperfusion (I/R) is a relevant clinical condition. Acetylcholine (ACh) and the α7 nicotinic ACh receptor (nAChRα-7) are involved in the control of inflammation. Mice with reduced levels of the vesicular ACh transporter (VAChT), a protein responsible for controlling ACh release, were used to test the involvement of cholinergic signaling in lung inflammation due to intestinal I/R. Female mice with reduced levels of VAChT (VAChT-KD
HOM ) or wild-type littermate controls (WT) were submitted to intestinal I/R followed by 2 h of reperfusion. Mortality, vascular permeability, and recruitment of inflammatory cells into the lung were investigated. Parts of mice were submitted to ovariectomy (OVx) to study the effect of sex hormones or treated with PNU-282,987 (nAChRα-7 agonist). A total of 43.4% of VAChT-KDHOM -I/R mice died in the reperfusion period compared to 5.2% of WT I/R mice. The I/R increased lung inflammation in both genotypes. In VAChT-KDHOM mice, I/R increased vascular permeability and decreased the release of cytokines in the lung compared to WT I/R mice. Ovariectomy reduced lung inflammation and permeability compared to non-OVx, but it did not avoid mortality in VAChT-KDHOM -I/R mice. PNU treatment reduced lung permeability, increased the release of proinflammatory cytokines and the myeloperoxidase activity in the lungs, and prevented the increased mortality observed in VAChT-KDHOM mice. Cholinergic signaling is an important component of the lung protector response against intestinal I/R injury. Decreased cholinergic signaling seems to increase pulmonary edema and dysfunctional cytokine release that increased mortality, which can be prevented by increasing activation of nAChRα-7., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)- Published
- 2021
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22. Acute Lung Injury in Cholinergic-Deficient Mice Supports Anti-Inflammatory Role of α7 Nicotinic Acetylcholine Receptor.
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Pinheiro NM, Banzato R, Tibério I, Prado MAM, Prado VF, Hamouda AK, and Prado CM
- Subjects
- Acute Lung Injury chemically induced, Acute Lung Injury metabolism, Acute Lung Injury pathology, Animals, Cytokines metabolism, Male, Mice, Nicotinic Agonists pharmacology, Pneumonia etiology, Pneumonia metabolism, Pneumonia pathology, Vesicular Acetylcholine Transport Proteins genetics, alpha7 Nicotinic Acetylcholine Receptor genetics, Acute Lung Injury drug therapy, Anti-Inflammatory Agents pharmacology, Benzamides pharmacology, Bridged Bicyclo Compounds pharmacology, Cholinergic Agents metabolism, Pneumonia prevention & control, Vesicular Acetylcholine Transport Proteins metabolism, alpha7 Nicotinic Acetylcholine Receptor metabolism
- Abstract
(1) Background: The lung cholinergic pathway is important for controlling pulmonary inflammation in acute lung injury, a condition that is characterized by a sudden onset and intense inflammation. This study investigated changes in the expression levels of nicotinic and muscarinic acetylcholine receptors (nAChR and mAChR) in the lung during acute lung injury. (2) Methods: acute lung injury (ALI) was induced in wild-type and cholinergic-deficient (VAChT-KD
HOM ) mice using intratracheal lipopolysaccharide (LPS) instillation with or without concurrent treatment with nicotinic ligands. Bronchoalveolar lavage fluid was collected to evaluate markers of inflammation, and then the lung was removed and processed for isolation of membrane fraction and determination of acetylcholine receptors level using radioligand binding assays. (3) Results: LPS-induced increase in lung inflammatory markers (e.g., neutrophils and IL-1β) was significantly higher in VAChT-KDHOM than wild-type mice. In contrast, LPS treatment resulted in a significant increase in lung's α7 nicotinic receptor level in wild-type, but not in VAChT-KDHOM mice. However, treatment with PNU 282987, a selective α7 nicotinic receptor agonist, restored VAChT-KDHOM mice's ability to increase α7 nicotinic receptor levels in response to LPS-induced acute lung injury and reduced lung inflammation. LPS also increased muscarinic receptors level in VAChT-KDHOM mice, and PNU 282987 treatment reduced this response. (4) Conclusions: Our data indicate that the anti-inflammatory effects of the lung cholinergic system involve an increase in the level of α7 nicotinic receptors. Pharmacological agents that increase the expression or the function of lung α7 nicotinic receptors have potential clinical uses for treating acute lung injury.- Published
- 2021
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23. Hsp90 and its co-chaperone Sti1 control TDP-43 misfolding and toxicity.
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Lin LT, Razzaq A, Di Gregorio SE, Hong S, Charles B, Lopes MH, Beraldo F, Prado VF, Prado MAM, and Duennwald ML
- Subjects
- Animals, Binding Sites, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Female, HSP90 Heat-Shock Proteins genetics, HeLa Cells, Humans, Inclusion Bodies, Mice, Mice, Inbred C57BL, Mice, Knockout, Neurons metabolism, Neurons pathology, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae growth & development, TDP-43 Proteinopathies etiology, Apoptosis, DNA-Binding Proteins chemistry, HSP90 Heat-Shock Proteins metabolism, Heat-Shock Proteins physiology, Protein Folding, Saccharomyces cerevisiae metabolism, TDP-43 Proteinopathies pathology
- Abstract
Protein misfolding is a central feature of most neurodegenerative diseases. Molecular chaperones can modulate the toxicity associated with protein misfolding, but it remains elusive which molecular chaperones and co-chaperones interact with specific misfolded proteins. TDP-43 misfolding and inclusion formation are a hallmark of amyotrophic lateral sclerosis (ALS) and other neurodegenerative diseases. Using yeast and mammalian neuronal cells we find that Hsp90 and its co-chaperone Sti1 have the capacity to alter TDP-43 misfolding, inclusion formation, aggregation, and cellular toxicity. Our data also demonstrate that impaired Hsp90 function sensitizes cells to TDP-43 toxicity and that Sti1 specifically interacts with and strongly modulates TDP-43 toxicity in a dose-dependent manner. Our study thus uncovers a previously unrecognized tie between Hsp90, Sti1, TDP-43 misfolding, and cellular toxicity., (© 2021 Federation of American Societies for Experimental Biology.)
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- 2021
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24. Increased Cholinergic Tone Causes Pre-synaptic Neuromuscular Degeneration and is Associated with Impaired Diaphragm Function.
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Magalhães-Gomes MPS, Camargos W, Valadão PAC, Garcias RS, Rodrigues HA, Andrade JN, Teixeira VP, Naves LA, Cavalcante WLG, Gallaci M, Guatimosim S, Prado VF, Prado MAM, and Guatimosim C
- Subjects
- Animals, Cholinergic Agents, Mice, Muscle, Skeletal metabolism, Neuromuscular Junction metabolism, Synaptic Transmission, Vesicular Acetylcholine Transport Proteins metabolism, DNA Copy Number Variations, Diaphragm metabolism
- Abstract
In vertebrates, muscle activity is dependent on acetylcholine (ACh) released from neuromuscular junctions (NMJs), and changes in cholinergic neurotransmission are linked to a variety of neuromuscular diseases, including congenital myasthenic syndromes (CMS). The storage and release of ACh depends on the activity of the Vesicular Acetylcholine Transporter (VAChT), a rate-limiting step for cholinergic neurotransmission whose loss of function mutations was shown to cause human congenital myasthenia. However, we know much less about increased VAChT activity, due to copy number variations, for example. Therefore, here we investigated the impact of increased VAChT expression and consequently ACh levels at the synaptic cleft of the diaphragm NMJs. We analyzed structure and function of nerve and muscles from a mouse model of cholinergic hyperfunction (ChAT-ChR2-EYFP) with increased expression of VAChT. Our results showed a significant increase of ACh released under evoked stimuli. However, we observed deleterious changes in synaptic vesicles cycle (impaired endocytosis and decrease in vesicles number), together with structural alterations of NMJs. Interestingly, ultrastructure analyses showed that synaptic vesicles from ChAT-ChR2-EYFP mice NMJs were larger, which might be related to increased ACh load. We also observed that these larger synaptic vesicles were less rounded in comparison with control. Finally, we showed that ChAT-ChR2-EYFP mice NMJs have compromised safety factor, possible due to the structural alterations we described. These findings reveal that physiological cholinergic activity is important to maintain the structure and function of the neuromuscular system and help to understand some of the neuromuscular adverse effects experienced by chronically increased NMJ neurotransmission, such as individuals treated with cholinesterase inhibitors., (Copyright © 2021 IBRO. All rights reserved.)
- Published
- 2021
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25. Forebrain Acetylcholine Modulates Isoflurane and Ketamine Anesthesia in Adult Mice.
- Author
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Leung LS, Chu L, Prado MAM, and Prado VF
- Subjects
- Animals, Male, Mice, Mice, Knockout, Models, Animal, Acetylcholine pharmacology, Anesthetics, Inhalation pharmacology, Isoflurane pharmacology, Ketamine pharmacology, Prosencephalon drug effects
- Abstract
Background: Cholinergic drugs are known to modulate general anesthesia, but anesthesia responses in acetylcholine-deficient mice have not been studied. It was hypothesized that mice with genetic deficiency of forebrain acetylcholine show increased anesthetic sensitivity to isoflurane and ketamine and decreased gamma-frequency brain activity., Methods: Male adult mice with heterozygous knockdown of vesicular acetylcholine transporter in the brain or homozygous knockout of the transporter in the basal forebrain were compared with wild-type mice. Hippocampal and frontal cortical electrographic activity and righting reflex were studied in response to isoflurane and ketamine doses., Results: The loss-of-righting-reflex dose for isoflurane was lower in knockout (mean ± SD, 0.76 ± 0.08%, n = 18, P = 0.005) but not knockdown (0.78 ± 0.07%, n = 24, P = 0.021), as compared to wild-type mice (0.83 ± 0.07%, n = 23), using a significance criterion of P = 0.017 for three planned comparisons. Loss-of-righting-reflex dose for ketamine was lower in knockout (144 ± 39 mg/kg, n = 14, P = 0.006) but not knockdown (162 ± 32 mg/kg, n = 20, P = 0.602) as compared to wild-type mice (168 ± 24 mg/kg, n = 21). Hippocampal high-gamma (63 to 100 Hz) power after isoflurane was significantly lower in knockout and knockdown mice compared to wild-type mice (isoflurane-dose and mouse-group interaction effect, F[8,56] = 2.87, P = 0.010; n = 5 to 6 mice per group). Hippocampal high-gamma power after ketamine was significantly lower in both knockout and knockdown mice when compared to wild-type mice (interaction effect F[2,13] = 6.06, P = 0.014). The change in frontal cortical gamma power with isoflurane or ketamine was not statistically different among knockout, knockdown, and wild-type mice., Conclusions: These findings suggest that forebrain cholinergic neurons modulate behavioral sensitivity and hippocampal gamma activity during isoflurane and ketamine anesthesia., (Copyright © 2021, the American Society of Anesthesiologists, Inc. All Rights Reserved.)
- Published
- 2021
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26. The brain in flux: Genetic, physiologic, and therapeutic perspectives on transporters in the CNS.
- Author
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Hewett SJ, Prado VF, and Robinson MB
- Subjects
- Animals, Humans, Blood-Brain Barrier metabolism, Brain physiology, Membrane Transport Proteins genetics, Membrane Transport Proteins metabolism
- Abstract
Active and passive transporters constitute a gene family of approximately 2000 members. These proteins are required for import and export across the blood brain barrier, clearance of neurotransmitters, inter-cellular solute transfer, and transport across the membranes of subcellular organelles. Neurologic, neurodevelopmental, and psychiatric diseases have been linked to alterations in function and/or mutations in every one of these types of transporters, and many of the transporters are targeted by therapeutics. This is the 4th biennial special edition of Neurochemistry International that originates from a scientific meeting devoted to studies of transporters and their relationship to brain function and to neurodevelopmental, neurologic, and psychiatric disorders. This meeting provides the only international forum for the presentation and discussion of cutting-edge research on brain transporters covering fundamental aspects of transporter structure, function, and trafficking. Scientists describe the novel approaches being used to link this information to physiology/circuit function and behavior. The meeting also addresses translational topics surrounding mouse models of brain transporter disorders, novel human brain disorders arising from transporter mutations, and innovative therapeutic approaches centered on modification of transporter function. This special issue includes a sampling of review articles that address timely questions of the field and several primary research articles., (Copyright © 2021. Published by Elsevier Ltd.)
- Published
- 2021
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27. New frontiers in translational research: Touchscreens, open science, and the mouse translational research accelerator platform.
- Author
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Sullivan JA, Dumont JR, Memar S, Skirzewski M, Wan J, Mofrad MH, Ansari HZ, Li Y, Muller L, Prado VF, Prado MAM, Saksida LM, and Bussey TJ
- Subjects
- Animals, Behavioral Research instrumentation, Citizen Science methods, Mice, Translational Science, Biomedical instrumentation, User-Computer Interface, Behavioral Research methods, Disease Models, Animal, Translational Science, Biomedical methods
- Abstract
Many neurodegenerative and neuropsychiatric diseases and other brain disorders are accompanied by impairments in high-level cognitive functions including memory, attention, motivation, and decision-making. Despite several decades of extensive research, neuroscience is little closer to discovering new treatments. Key impediments include the absence of validated and robust cognitive assessment tools for facilitating translation from animal models to humans. In this review, we describe a state-of-the-art platform poised to overcome these impediments and improve the success of translational research, the Mouse Translational Research Accelerator Platform (MouseTRAP), which is centered on the touchscreen cognitive testing system for rodents. It integrates touchscreen-based tests of high-level cognitive assessment with state-of-the art neurotechnology to record and manipulate molecular and circuit level activity in vivo in animal models during human-relevant cognitive performance. The platform also is integrated with two Open Science platforms designed to facilitate knowledge and data-sharing practices within the rodent touchscreen community, touchscreencognition.org and mousebytes.ca. Touchscreencognition.org includes the Wall, showcasing touchscreen news and publications, the Forum, for community discussion, and Training, which includes courses, videos, SOPs, and symposia. To get started, interested researchers simply create user accounts. We describe the origins of the touchscreen testing system, the novel lines of research it has facilitated, and its increasingly widespread use in translational research, which is attributable in part to knowledge-sharing efforts over the past decade. We then identify the unique features of MouseTRAP that stand to potentially revolutionize translational research, and describe new initiatives to partner with similar platforms such as McGill's M3 platform (m3platform.org)., (© 2020 International Behavioural and Neural Genetics Society and John Wiley & Sons Ltd.)
- Published
- 2021
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28. Cholinergic dysfunction in the dorsal striatum promotes habit formation and maladaptive eating.
- Author
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Favier M, Janickova H, Justo D, Kljakic O, Runtz L, Natsheh JY, Pascoal TA, Germann J, Gallino D, Kang JI, Meng XQ, Antinora C, Raulic S, Jacobsen JP, Moquin L, Vigneault E, Gratton A, Caron MG, Duriez P, Brandon MP, Neto PR, Chakravarty MM, Herzallah MM, Gorwood P, Prado MA, Prado VF, and El Mestikawy S
- Subjects
- Adult, Animals, Donepezil pharmacology, Feeding Behavior drug effects, Feeding and Eating Disorders drug therapy, Feeding and Eating Disorders genetics, Feeding and Eating Disorders physiopathology, Female, Humans, Levodopa pharmacology, Male, Mice, Mice, Knockout, Middle Aged, Vesicular Acetylcholine Transport Proteins genetics, Vesicular Acetylcholine Transport Proteins metabolism, Acetylcholine metabolism, Corpus Striatum metabolism, Corpus Striatum physiopathology, Feeding and Eating Disorders metabolism, Glutamic Acid metabolism, Interneurons metabolism
- Abstract
Dysregulation of habit formation has been recently proposed as pivotal to eating disorders. Here, we report that a subset of patients suffering from restrictive anorexia nervosa have enhanced habit formation compared with healthy controls. Habit formation is modulated by striatal cholinergic interneurons. These interneurons express vesicular transporters for acetylcholine (VAChT) and glutamate (VGLUT3) and use acetylcholine/glutamate cotransmission to regulate striatal functions. Using mice with genetically silenced VAChT (VAChT conditional KO, VAChTcKO) or VGLUT3 (VGLUT3cKO), we investigated the roles that acetylcholine and glutamate released by cholinergic interneurons play in habit formation and maladaptive eating. Silencing glutamate favored goal-directed behaviors and had no impact on eating behavior. In contrast, VAChTcKO mice were more prone to habits and maladaptive eating. Specific deletion of VAChT in the dorsomedial striatum of adult mice was sufficient to phenocopy maladaptive eating behaviors of VAChTcKO mice. Interestingly, VAChTcKO mice had reduced dopamine release in the dorsomedial striatum but not in the dorsolateral striatum. The dysfunctional eating behavior of VAChTcKO mice was alleviated by donepezil and by l-DOPA, confirming an acetylcholine/dopamine deficit. Our study reveals that loss of acetylcholine leads to a dopamine imbalance in striatal compartments, thereby promoting habits and vulnerability to maladaptive eating in mice.
- Published
- 2020
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29. Infiltrating Hematogenous Macrophages Aggregate Around β-Amyloid Plaques in an Age- and Sex-Dependent Manner in a Mouse Model of Alzheimer Disease.
- Author
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Kozyrev N, Albers S, Yang J, Prado VF, Prado MAM, Fonseca GJ, Rylett RJ, and Dekaban GA
- Subjects
- Alzheimer Disease immunology, Amyloid beta-Peptides immunology, Amyloid beta-Peptides metabolism, Animals, Brain immunology, Disease Models, Animal, Female, Inflammation immunology, Inflammation pathology, Male, Mice, Mice, Transgenic, Plaque, Amyloid immunology, Alzheimer Disease pathology, Brain pathology, Macrophages pathology, Plaque, Amyloid pathology
- Abstract
β-Amyloid (Aβ) plaques can trigger chronic inflammation in the cellular environment that recruits infiltrating macrophages during the course of Alzheimer disease (AD). Activated macrophages release pro-inflammatory cytokines that increase neurotoxicity associated with AD. A major impediment to investigating neuroinflammation involving macrophage activity is the inability to discriminate resident microglial macrophages (mMϕ) from hematogenous macrophages (hMϕ), as they are morphologically and phenotypically similar when activated. To distinguish between mMϕ and hMϕ and to determine their respective roles in chronic inflammation associated with the progression of amyloidosis, we used lys-EGFP-ki transgenic mice that express enhanced green fluorescent protein in hMϕ, but not in mMϕ. These mice were crossed with 5XFAD mice. The offspring demonstrated robust AD pathology and enabled visual discrimination of mMϕ from hMϕ. Mutant mice demonstrated robust increases in Aβ1-42, area of Aβ plaques, gliosis and deficits in spatial learning by age 5 months. The time-course of Aβ accumulation, paralleled by the accumulation of hMϕ around Aβ plaques, was more robust in female compared with male mice and preceded behavioral changes. Thus, the accumulation of infiltrating hMϕ around Aβ plaques was age- and sex-dependent and preceded cognitive impairment., (© 2020 American Association of Neuropathologists, Inc. All rights reserved.)
- Published
- 2020
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30. An optimized acetylcholine sensor for monitoring in vivo cholinergic activity.
- Author
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Jing M, Li Y, Zeng J, Huang P, Skirzewski M, Kljakic O, Peng W, Qian T, Tan K, Zou J, Trinh S, Wu R, Zhang S, Pan S, Hires SA, Xu M, Li H, Saksida LM, Prado VF, Bussey TJ, Prado MAM, Chen L, Cheng H, and Li Y
- Subjects
- Animals, Animals, Genetically Modified, Behavior, Animal physiology, Cholinergic Agents pharmacology, Drosophila genetics, Drosophila metabolism, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, HEK293 Cells, Humans, Mice, Mushroom Bodies metabolism, Neurons metabolism, Olfactory Cortex metabolism, Receptor, Muscarinic M3 genetics, Receptor, Muscarinic M3 metabolism, Somatosensory Cortex metabolism, Acetylcholine metabolism, Biosensing Techniques methods, Brain metabolism
- Abstract
The ability to directly measure acetylcholine (ACh) release is an essential step toward understanding its physiological function. Here we optimized the GRAB
ACh (GPCR-activation-based ACh) sensor to achieve substantially improved sensitivity in ACh detection, as well as reduced downstream coupling to intracellular pathways. The improved version of the ACh sensor retains the subsecond response kinetics, physiologically relevant affinity and precise molecular specificity for ACh of its predecessor. Using this sensor, we revealed compartmental ACh signals in the olfactory center of transgenic flies in response to external stimuli including odor and body shock. Using fiber photometry recording and two-photon imaging, our ACh sensor also enabled sensitive detection of single-trial ACh dynamics in multiple brain regions in mice performing a variety of behaviors.- Published
- 2020
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31. Effects of VAChT reduction and α7nAChR stimulation by PNU-282987 in lung inflammation in a model of chronic allergic airway inflammation.
- Author
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Pinheiro NM, Miranda CJCP, Santana FR, Bittencourt-Mernak M, Arantes-Costa FM, Olivo C, Perini A, Festa S, Caperuto LC, Tibério IFLC, Prado MAM, Martins MA, Prado VF, and Prado CM
- Subjects
- Airway Remodeling, Allergens, Animals, Asthma etiology, Benzamides pharmacology, Bridged Bicyclo Compounds pharmacology, Bronchoalveolar Lavage Fluid cytology, Bronchoalveolar Lavage Fluid immunology, Chronic Disease, Cytokines immunology, Disease Models, Animal, Inflammation etiology, Inflammation immunology, Leukocyte Count, Lung drug effects, Lung immunology, Lung pathology, Male, Mice, Inbred BALB C, Mice, Knockout, Ovalbumin, STAT3 Transcription Factor antagonists & inhibitors, Suppressor of Cytokine Signaling 3 Protein antagonists & inhibitors, Vesicular Acetylcholine Transport Proteins genetics, alpha7 Nicotinic Acetylcholine Receptor agonists, Asthma immunology, Vesicular Acetylcholine Transport Proteins deficiency, alpha7 Nicotinic Acetylcholine Receptor immunology
- Abstract
The cholinergic anti-inflammatory pathway has been shown to regulate lung inflammation and cytokine release in acute models of inflammation, mainly via α7 nicotinic receptor (α7nAChR). We aimed to evaluate the role of endogenous acetylcholine in chronic allergic airway inflammation in mice and the effects of therapeutic nAChR stimulation in this model. We first evaluated lung inflammation and remodeling on knock-down mice with 65% of vesicular acetylcholine transport (VAChT) gene reduction (KDVAChT) and wild-type(WT) controls that were subcutaneously sensitized and then inhaled with ovalbumin(OVA). We then evaluated the effects of PNU-282987(0.5-to-2mg/kg),(α7nAChR agonist) treatment in BALB/c male mice intraperitoneal sensitized and then inhaled with OVA. Another OVA-sensitized-group was treated with PNU-282987 plus Methyllycaconitine (MLA,1 mg/kg, α7nAChR antagonist) to confirm that the effects observed by PNU were due to α7nAChR. We showed that KDVAChT-OVA mice exhibit exacerbated airway inflammation when compared to WT-OVA mice. In BALB/c, PNU-282987 treatment reduced the number of eosinophils in the blood, BAL fluid, and around airways, and also decreased pulmonary levels of IL-4,IL-13,IL-17, and IgE in the serum of OVA-exposed mice. MLA pre-treatment abolished all the effects of PNU-282987. Additionally, we showed that PNU-282987 inhibited STAT3-phosphorylation and reduced SOCS3 expression in the lung. These data indicate that endogenous cholinergic tone is important to control allergic airway inflammation in a murine model. Moreover, α7nAChR is involved in the control of eosinophilic inflammation and airway remodeling, possibly via inhibition of STAT3/SOCS3 pathways. Together these data suggest that cholinergic anti-inflammatory system mainly α7nAChR should be further considered as a therapeutic target in asthma., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
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32. Increased levels of Stress-inducible phosphoprotein-1 accelerates amyloid-β deposition in a mouse model of Alzheimer's disease.
- Author
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Lackie RE, Marques-Lopes J, Ostapchenko VG, Good S, Choy WY, van Oosten-Hawle P, Pasternak SH, Prado VF, and Prado MAM
- Subjects
- Alzheimer Disease metabolism, Animals, Brain metabolism, Brain pathology, Caenorhabditis elegans, Caenorhabditis elegans Proteins metabolism, Humans, Mice, Nuclear Proteins metabolism, Alzheimer Disease pathology, Amyloid beta-Peptides metabolism, Disease Models, Animal, Heat-Shock Proteins metabolism, Plaque, Amyloid metabolism
- Abstract
Molecular chaperones and co-chaperones, which are part of the protein quality control machinery, have been shown to regulate distinct aspects of Alzheimer's Disease (AD) pathology in multiple ways. Notably, the co-chaperone STI1, which presents increased levels in AD, can protect mammalian neurons from amyloid-β toxicity in vitro and reduced STI1 levels worsen Aβ toxicity in C. elegans. However, whether increased STI1 levels can protect neurons in vivo remains unknown. We determined that overexpression of STI1 and/or Hsp90 protected C. elegans expressing Aβ
(3-42) against Aβ-mediated paralysis. Mammalian neurons were also protected by elevated levels of endogenous STI1 in vitro, and this effect was mainly due to extracellular STI1. Surprisingly, in the 5xFAD mouse model of AD, by overexpressing STI1, we find increased amyloid burden, which amplifies neurotoxicity and worsens spatial memory deficits in these mutants. Increased levels of STI1 disturbed the expression of Aβ-regulating enzymes (BACE1 and MMP-2), suggesting potential mechanisms by which amyloid burden is increased in mice. Notably, we observed that STI1 accumulates in dense-core AD plaques in both 5xFAD mice and human brain tissue. Our findings suggest that elevated levels of STI1 contribute to Aβ accumulation, and that STI1 is deposited in AD plaques in mice and humans. We conclude that despite the protective effects of STI1 in C. elegans and in mammalian cultured neurons, in vivo, the predominant effect of elevated STI1 is deleterious in AD.- Published
- 2020
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33. Chronic hM3Dq signaling in microglia ameliorates neuroinflammation in male mice.
- Author
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Binning W, Hogan-Cann AE, Yae Sakae D, Maksoud M, Ostapchenko V, Al-Onaizi M, Matovic S, Lu WY, Prado MAM, Inoue W, and Prado VF
- Subjects
- Acetylcholine, Animals, Male, Mice, Neurons, Receptors, G-Protein-Coupled, Signal Transduction, Clozapine pharmacology, Microglia
- Abstract
Microglia express muscarinic G protein-coupled receptors (GPCRs) that sense cholinergic activity and are activated by acetylcholine to potentially regulate microglial functions. Knowledge about how distinct types of muscarinic GPCR signaling regulate microglia function in vivo is still poor, partly due to the fact that some of these receptors are also present in astrocytes and neurons. We generated mice expressing the hM3Dq Designer Receptor Exclusively Activated by Designer Drugs (DREADD) selectively in microglia to investigate the role of muscarinic M3Gq-linked signaling. We show that activation of hM3Dq using clozapine N-oxide (CNO) elevated intracellular calcium levels and increased phagocytosis of FluoSpheres by microglia in vitro. Interestingly, whereas acute treatment with CNO increased synthesis of cytokine mRNA, chronic treatment attenuated LPS-induced cytokine mRNA changes in the brain. No effect of CNO on cytokine expression was observed in DREADD-negative mice. Interestingly, CNO activation of M3Dq in microglia was able to attenuate LPS-mediated decrease in social interactions. These results suggest that chronic activation of M3 muscarinic receptors (the hM3Dq progenitor) in microglia, and potentially other Gq-coupled GPCRs, can trigger an inflammatory-like response that preconditions microglia to decrease their response to further immunological challenges. Our results indicate that hM3Dq can be a useful tool to modulate neuroinflammation and study microglial immunological memory in vivo, which may be applicable for manipulations of neuroinflammation in neurodegenerative and psychiatric diseases., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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34. Modulation of hippocampal neuronal resilience during aging by the Hsp70/Hsp90 co-chaperone STI1.
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Lackie RE, Razzaq AR, Farhan SMK, Qiu LR, Moshitzky G, Beraldo FH, Lopes MH, Maciejewski A, Gros R, Fan J, Choy WY, Greenberg DS, Martins VR, Duennwald ML, Lerch JP, Soreq H, Prado VF, and Prado MAM
- Subjects
- Adaptation, Physiological physiology, Aging genetics, Animals, Embryonic Stem Cells metabolism, Gene Knockout Techniques methods, HSP70 Heat-Shock Proteins genetics, HSP90 Heat-Shock Proteins genetics, Heat-Shock Proteins genetics, Hippocampus cytology, Humans, Male, Mice, Mice, Inbred C57BL, Molecular Chaperones genetics, Neurons metabolism, Aging metabolism, HSP70 Heat-Shock Proteins metabolism, HSP90 Heat-Shock Proteins metabolism, Heat-Shock Proteins deficiency, Hippocampus metabolism, Molecular Chaperones metabolism
- Abstract
Chaperone networks are dysregulated with aging, but whether compromised Hsp70/Hsp90 chaperone function disturbs neuronal resilience is unknown. Stress-inducible phosphoprotein 1 (STI1; STIP1; HOP) is a co-chaperone that simultaneously interacts with Hsp70 and Hsp90, but whose function in vivo remains poorly understood. We combined in-depth analysis of chaperone genes in human datasets, analysis of a neuronal cell line lacking STI1 and of a mouse line with a hypomorphic Stip1 allele to investigate the requirement for STI1 in aging. Our experiments revealed that dysfunctional STI1 activity compromised Hsp70/Hsp90 chaperone network and neuronal resilience. The levels of a set of Hsp90 co-chaperones and client proteins were selectively affected by reduced levels of STI1, suggesting that their stability depends on functional Hsp70/Hsp90 machinery. Analysis of human databases revealed a subset of co-chaperones, including STI1, whose loss of function is incompatible with life in mammals, albeit they are not essential in yeast. Importantly, mice expressing a hypomorphic STI1 allele presented spontaneous age-dependent hippocampal neurodegeneration and reduced hippocampal volume, with consequent spatial memory deficit. We suggest that impaired STI1 function compromises Hsp70/Hsp90 chaperone activity in mammals and can by itself cause age-dependent hippocampal neurodegeneration in mice. Cover Image for this issue: doi: 10.1111/jnc.14749., (© 2019 International Society for Neurochemistry.)
- Published
- 2020
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35. Optimizing Nervous System-Specific Gene Targeting with Cre Driver Lines: Prevalence of Germline Recombination and Influencing Factors.
- Author
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Luo L, Ambrozkiewicz MC, Benseler F, Chen C, Dumontier E, Falkner S, Furlanis E, Gomez AM, Hoshina N, Huang WH, Hutchison MA, Itoh-Maruoka Y, Lavery LA, Li W, Maruo T, Motohashi J, Pai EL, Pelkey KA, Pereira A, Philips T, Sinclair JL, Stogsdill JA, Traunmüller L, Wang J, Wortel J, You W, Abumaria N, Beier KT, Brose N, Burgess HA, Cepko CL, Cloutier JF, Eroglu C, Goebbels S, Kaeser PS, Kay JN, Lu W, Luo L, Mandai K, McBain CJ, Nave KA, Prado MAM, Prado VF, Rothstein J, Rubenstein JLR, Saher G, Sakimura K, Sanes JR, Scheiffele P, Takai Y, Umemori H, Verhage M, Yuzaki M, Zoghbi HY, Kawabe H, and Craig AM
- Subjects
- Animals, Female, Genes, Reporter, Germ Cells, Male, Mice, Mice, Transgenic, Mosaicism, Gene Targeting methods, Integrases genetics, Neurons metabolism, Oocytes metabolism, Recombination, Genetic genetics, Spermatozoa metabolism
- Abstract
The Cre-loxP system is invaluable for spatial and temporal control of gene knockout, knockin, and reporter expression in the mouse nervous system. However, we report varying probabilities of unexpected germline recombination in distinct Cre driver lines designed for nervous system-specific recombination. Selective maternal or paternal germline recombination is showcased with sample Cre lines. Collated data reveal germline recombination in over half of 64 commonly used Cre driver lines, in most cases with a parental sex bias related to Cre expression in sperm or oocytes. Slight differences among Cre driver lines utilizing common transcriptional control elements affect germline recombination rates. Specific target loci demonstrated differential recombination; thus, reporters are not reliable proxies for another locus of interest. Similar principles apply to other recombinase systems and other genetically targeted organisms. We hereby draw attention to the prevalence of germline recombination and provide guidelines to inform future research for the neuroscience and broader molecular genetics communities., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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36. Striatal Acetylcholine Helps to Preserve Functional Outcomes in a Mouse Model of Stroke.
- Author
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Goncalves DF, Guzman MS, Gros R, Massensini AR, Bartha R, Prado VF, and Prado MAM
- Subjects
- Acetylcholine genetics, Animals, Corpus Striatum pathology, Disease Models, Animal, Infarction, Middle Cerebral Artery genetics, Infarction, Middle Cerebral Artery pathology, Male, Mice, Mice, Knockout, Mice, Transgenic, Stroke genetics, Stroke pathology, Vesicular Acetylcholine Transport Proteins deficiency, Vesicular Acetylcholine Transport Proteins genetics, Acetylcholine metabolism, Corpus Striatum metabolism, Infarction, Middle Cerebral Artery metabolism, Stroke metabolism
- Abstract
Acetylcholine (ACh) has been suggested to facilitate plasticity and improve functional recovery after different types of brain lesions. Interestingly, numerous studies have shown that striatal cholinergic interneurons are relatively resistant to acute ischemic insults, but whether ACh released by these neurons enhances functional recovery after stroke is unknown. We investigated the role of endogenous striatal ACh in stroke lesion volume and functional outcomes following middle cerebral artery occlusion to induce focal ischemia in striatum-selective vesicular acetylcholine transporter-deficient mice (stVAChT-KO). As transporter expression is almost completely eliminated in the striatum of stVAChT-KO mice, ACh release is nearly abolished in this area. Conversely, in other brain areas, VAChT expression and ACh release are preserved. Our results demonstrate a larger infarct size after ischemic insult in stVAChT-KO mice, with more pronounced functional impairments and increased mortality than in littermate controls. These changes are associated with increased activation of GSK-3, decreased levels of β-catenin, and a higher permeability of the blood-brain barrier in mice with loss of VAChT in striatum neurons. These results support a framework in which endogenous ACh secretion originating from cholinergic interneurons in the striatum helps to protect brain tissue against ischemia-induced damage and facilitates brain recovery by supporting blood-brain barrier function.
- Published
- 2020
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37. MouseBytes, an open-access high-throughput pipeline and database for rodent touchscreen-based cognitive assessment.
- Author
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Beraldo FH, Palmer D, Memar S, Wasserman DI, Lee WV, Liang S, Creighton SD, Kolisnyk B, Cowan MF, Mels J, Masood TS, Fodor C, Al-Onaizi MA, Bartha R, Gee T, Saksida LM, Bussey TJ, Strother SS, Prado VF, Winters BD, and Prado MA
- Subjects
- Alzheimer Disease, Animals, Behavior, Animal, Choice Behavior, Databases, Factual, Disease Models, Animal, Female, Learning physiology, Male, Memory physiology, Mice, Neuropsychological Tests, Reproducibility of Results, Software, Cognition physiology, Laboratory Animal Science instrumentation, Laboratory Animal Science methods, Rodentia genetics
- Abstract
Open Science has changed research by making data accessible and shareable, contributing to replicability to accelerate and disseminate knowledge. However, for rodent cognitive studies the availability of tools to share and disseminate data is scarce. Automated touchscreen-based tests enable systematic cognitive assessment with easily standardised outputs that can facilitate data dissemination. Here we present an integration of touchscreen cognitive testing with an open-access database public repository (mousebytes.ca), as well as a Web platform for knowledge dissemination (https://touchscreencognition.org). We complement these resources with the largest dataset of age-dependent high-level cognitive assessment of mouse models of Alzheimer's disease, expanding knowledge of affected cognitive domains from male and female mice of three strains. We envision that these new platforms will enhance sharing of protocols, data availability and transparency, allowing meta-analysis and reuse of mouse cognitive data to increase the replicability/reproducibility of datasets., Competing Interests: FB, DP, SM, DW, WL, SL, SC, BK, MC, JM, TM, CF, MA, RB, TG, SS, VP, BW, MP No competing interests declared, LS Lisa Saksida has established a series of targeted cognitive tests for animals, administered via touchscreen within a custom environment known as the “Bussey-Saksida touchscreen chamber”. Cambridge Enterprise, the technology transfer office of the University of Cambridge, supported commercialization of the Bussey-Saksida chamber, culminating in a license to Campden Instruments. Any financial compensation received from commercialization of the technology is fully invested in further touchscreen development and/or maintenance, TB Tim Bussey has established a series of targeted cognitive tests for animals, administered via touchscreen within a custom environment known as the “Bussey-Saksida touchscreen chamber”. Cambridge Enterprise, the technology transfer office of the University of Cambridge, supported commercialization of the Bussey-Saksida chamber, culminating in a license to Campden Instruments. Any financial compensation received from commercialization of the technology is fully invested in further touchscreen development and/or maintenance, (© 2019, Beraldo et al.)
- Published
- 2019
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38. Evaluation of the neuromuscular junction in a middle-aged mouse model of congenital myasthenic syndrome.
- Author
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Leite Schetino LP, de Castro Fonseca M, Magalhães Gomes MPS, Costa Valadão PA, de Camargo WL, Rodrigues HA, Andrade JN, Arantes-Costa FM, Naves LA, Prado CM, Prado VF, Prado MAM, and Guatimosim C
- Subjects
- Acetylcholine metabolism, Aging metabolism, Animals, Diaphragm metabolism, Diaphragm physiopathology, Disease Models, Animal, Endocytosis, Excitatory Postsynaptic Potentials physiology, Exocytosis, Gene Knockdown Techniques, Mice, Microscopy, Confocal, Microscopy, Electron, Transmission, Motor Endplate, Myasthenic Syndromes, Congenital genetics, Myasthenic Syndromes, Congenital metabolism, Myasthenic Syndromes, Congenital physiopathology, Neuromuscular Junction metabolism, Neuromuscular Junction physiopathology, Respiratory Mechanics physiology, Synaptic Transmission, Synaptic Vesicles metabolism, Vesicular Acetylcholine Transport Proteins genetics, Aging pathology, Diaphragm ultrastructure, Myasthenic Syndromes, Congenital pathology, Neuromuscular Junction ultrastructure, Synaptic Vesicles ultrastructure
- Abstract
Introduction: Reduced expression of the vesicular acetylcholine transporter (VAChT) leads to changes in the distribution and shape of synaptic vesicles (SVs) at neuromuscular junctions (NMJs), suggesting vesicular acetylcholine (ACh) as a key component of synaptic structure and function. It is poorly understood how long-term changes in cholinergic transmission contribute to age- and disease-related degeneration in the motor system., Methods: In this study we performed confocal imaging, electrophysiology, electron microscopy, and analyses of respiratory mechanics of the diaphragm NMJ components in 12-month-old wild-type (WT) and VAChTKD
HOM mice., Results: Diaphragms of NMJs of the VAChTKDHOM mice were similar to those in WT mice in number, colocalization, and fragmentation of pre-/postsynaptic components. However, they had increased spontaneous SV exocytosis, miniature endplate potential frequency, and diminished MEPP amplitude. No impairment in respiratory mechanics at rest was observed, probably due to the large neurotransmission safety factor of the diaphragm., Discussion: The present findings help us to understand the consequences of reduced ACh release at the NMJs during aging., (© 2019 Wiley Periodicals, Inc.)- Published
- 2019
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39. Selective decrease of cholinergic signaling from pedunculopontine and laterodorsal tegmental nuclei has little impact on cognition but markedly increases susceptibility to stress.
- Author
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Janickova H, Kljakic O, Rosborough K, Raulic S, Matovic S, Gros R, Saksida LM, Bussey TJ, Inoue W, Prado VF, and Prado MAM
- Subjects
- Animals, Attention, Corticosterone blood, Gene Expression Regulation, Vesicular Acetylcholine Transport Proteins genetics, Cognition physiology, Lateral Thalamic Nuclei physiology, Pedunculopontine Tegmental Nucleus physiology, Stress, Physiological
- Abstract
The pedunculopontine tegmental nucleus (PPT) and laterodorsal tegmental nucleus (LDT) are heterogeneous brainstem structures that contain cholinergic, glutamatergic, and GABAergic neurons. PPT/LDT neurons are suggested to modulate both cognitive and noncognitive functions, yet the extent to which acetylcholine (ACh) signaling from the PPT/LDT is necessary for normal behavior remains uncertain. We addressed this issue by using a mouse model in which PPT/LDT cholinergic signaling is highly decreased by selective deletion of the vesicular ACh transporter ( VAChT ) gene. This approach interferes exclusively with ACh signaling, leaving signaling by other neurotransmitters from PPT/LDT cholinergic neurons intact and sparing other cells. VAChT mutants were examined on different PPT/LDT-associated cognitive domains. Interestingly, VAChT mutants showed no attentional deficits and only minor cognitive flexibility impairments while presenting large deficiencies in both spatial and cued Morris water maze (MWM) tasks. Conversely, working spatial memory determined with the Y-maze and spatial memory measured with the Barnes maze were not affected, suggesting that deficits in MWM were unrelated to spatial memory abnormalities. Supporting this interpretation, VAChT mutants exhibited alterations in anxiety-like behavior and increased corticosterone levels after exposure to the MWM, suggesting altered stress response. Thus, PPT/LDT VAChT-mutant mice present little cognitive impairment per se , yet they exhibit increased susceptibility to stress, which may lead to performance deficits in more stressful conditions.-Janickova, H., Kljakic, O., Rosborough, K., Raulic, S., Matovic, S., Gros, R., Saksida, L. M., Bussey, T. J., Inoue, W., Prado, V. F., Prado, M. A. M. Selective decrease of cholinergic signaling from pedunculopontine and laterodorsal tegmental nuclei has little impact on cognition but markedly increases susceptibility to stress.
- Published
- 2019
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40. Forebrain Cholinergic Signaling Regulates Innate Immune Responses and Inflammation.
- Author
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Lehner KR, Silverman HA, Addorisio ME, Roy A, Al-Onaizi MA, Levine Y, Olofsson PS, Chavan SS, Gros R, Nathanson NM, Al-Abed Y, Metz CN, Prado VF, Prado MAM, Tracey KJ, and Pavlov VA
- Subjects
- Acetylcholine pharmacology, Animals, Anti-Inflammatory Agents pharmacology, Cholinergic Agonists pharmacology, Cholinesterase Inhibitors pharmacology, Cytokines blood, Cytokines immunology, Endotoxemia immunology, Endotoxemia metabolism, Galantamine pharmacology, Immunity, Innate, Inflammation immunology, Inflammation metabolism, Male, Mice, Inbred C57BL, Mice, Transgenic, Prosencephalon metabolism, Quinolines pharmacology, Receptor, Muscarinic M1 genetics, Receptor, Muscarinic M1 metabolism, Vesicular Acetylcholine Transport Proteins genetics, Prosencephalon immunology, Receptor, Muscarinic M1 immunology
- Abstract
The brain regulates physiological functions integral to survival. However, the insight into brain neuronal regulation of peripheral immune function and the neuromediator systems and pathways involved remains limited. Here, utilizing selective genetic and pharmacological approaches, we studied the role of forebrain cholinergic signaling in the regulation of peripheral immune function and inflammation. Forebrain-selective genetic ablation of acetylcholine release and vagotomy abolished the suppression of serum TNF by the centrally-acting cholinergic drug galantamine in murine endotoxemia. Selective stimulation of acetylcholine action on the M1 muscarinic acetylcholine receptor (M1 mAChR) by central administration of the positive allosteric modulator benzyl quinolone carboxylic acid (BQCA) suppressed serum TNF (TNFα) levels in murine endotoxemia. This effect was recapitulated by peripheral administration of the compound. BQCA also improved survival in murine endotoxemia and these effects were abolished in M1 mAChR knockout (KO) mice. Selective optogenetic stimulation of basal forebrain cholinergic neurons innervating brain regions with abundant M1 mAChR localization reduced serum TNF in endotoxemic mice. These findings reveal that forebrain cholinergic neurons regulate innate immune responses and inflammation, suggesting the possibility that in diseases associated with cholinergic dysfunction, including Alzheimer's disease this anti-inflammatory regulation can be impaired. These results also suggest novel anti-inflammatory approaches based on targeting forebrain cholinergic signaling in sepsis and other disorders characterized by immune dysregulation.
- Published
- 2019
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41. Vesicular acetylcholine transport deficiency potentiates some inflammatory responses induced by diesel exhaust particles.
- Author
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Santana FPR, Pinheiro NM, Bittencourt-Mernak MI, Perini A, Yoshizaki K, Macchione M, Saldiva PHN, Martins MA, Tibério IFLC, Prado MAM, Prado VF, and Prado CM
- Subjects
- Animals, Bronchoalveolar Lavage Fluid cytology, Cytokines genetics, Cytokines metabolism, Lung metabolism, Macrophages drug effects, Macrophages metabolism, Male, Mice, Parenchymal Tissue drug effects, Parenchymal Tissue metabolism, Pneumonia chemically induced, Pneumonia diagnosis, Vesicular Acetylcholine Transport Proteins deficiency, Vesicular Acetylcholine Transport Proteins metabolism, Lung drug effects, Vehicle Emissions toxicity, Vesicular Acetylcholine Transport Proteins genetics
- Abstract
Endogenous acetylcholine (ACh), which depends of the levels of vesicular ACh transport (VAChT) to be released, is the central mediator of the cholinergic anti-inflammatory system. ACh controls the release of cytokine in different models of inflammation. Diesel exhaust particles (DEP) are one of the major environmental pollutants produced in large quantity by automotive engines in urban center. DEP bind the lung parenchyma and induce inflammation. We evaluated whether cholinergic dysfunction worsens DEP-induced lung inflammation. Male mice with decreased ACh release due to reduced expression of VAChT (VAChT-KD mice) were submitted to DEP exposure for 30 days (3 mg/mL of DEP, once a day, five days a week) or saline. Pulmonary function and inflammation as well as extracellular matrix fiber deposition were evaluated. Additionally, airway and nasal epithelial mucus production were quantified. We found that DEP instillation worsened lung function and increased lung inflammation. Higher levels of mononuclear cells were observed in the peripheral blood of both wild-type (WT) and VAChT-KD mice. Also, both wild-type (WT) and VAChT-KD mice showed an increase in macrophages in bronchoalveolar lavage fluid (BALF) as well as increased expression of IL-4, IL-6, IL-13, TNF-α, and NF-κB in lung cells. The collagen fiber content in alveolar septa was also increased in both genotypes. On the other hand, we observed that granulocytes were increased only in VAChT-KD peripheral blood. Likewise, increased BALF lymphocytes and neutrophils as well as increased elastic fibers in alveolar septa, airway neutral mucus, and nasal epithelia acid mucus were observed only in VAChT-KD mice. The cytokines IL-4 and TNF-α were also higher in VAChT-KD mice compared with WT mice. In conclusion, decreased ability to release ACh exacerbates some of the lung alterations induced by DEP in mice, suggesting that VAChT-KD animals are more vulnerable to the effects of DEP in the lung., (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Published
- 2019
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42. Detection of Active Caspase-3 in Mouse Models of Stroke and Alzheimer's Disease with a Novel Dual Positron Emission Tomography/Fluorescent Tracer [ 68 Ga]Ga-TC3-OGDOTA.
- Author
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Ostapchenko VG, Snir J, Suchy M, Fan J, Cobb MR, Chronik BA, Kovacs M, Prado VF, Hudson RHE, Pasternak SH, Prado MAM, and Bartha R
- Subjects
- Animals, Cells, Cultured, Female, Kinetics, Male, Mice, Microscopy, Confocal, Alzheimer Disease diagnostic imaging, Caspase 3 metabolism, Gallium Radioisotopes chemistry, Positron-Emission Tomography methods, Stroke diagnostic imaging
- Abstract
Apoptosis is a feature of stroke and Alzheimer's disease (AD), yet there is no accepted method to detect or follow apoptosis in the brain in vivo. We developed a bifunctional tracer [
68 Ga]Ga-TC3-OGDOTA containing a cell-penetrating peptide separated from fluorescent Oregon Green and68 Ga-bound labels by the caspase-3 recognition peptide DEVD. We hypothesized that this design would allow [68 Ga]Ga-TC3-OGDOTA to accumulate in apoptotic cells. In vitro, Ga-TC3-OGDOTA labeled apoptotic neurons following exposure to camptothecin, oxygen-glucose deprivation, and β -amyloid oligomers. In vivo, PET showed accumulation of [68 Ga]Ga-TC3-OGDOTA in the brain of mouse models of stroke or AD. Optical clearing revealed colocalization of [68 Ga]Ga-TC3-OGDOTA and cleaved caspase-3 in brain cells. In stroke, [68 Ga]Ga-TC3-OGDOTA accumulated in neurons in the penumbra area, whereas in AD mice [68 Ga]Ga-TC3-OGDOTA was found in single cells in the forebrain and diffusely around amyloid plaques. In summary, this bifunctional tracer is selectively associated with apoptotic cells in vitro and in vivo in brain disease models and represents a novel tool for apoptosis detection that can be used in neurodegenerative diseases.- Published
- 2019
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43. Dissociable cognitive impairments in two strains of transgenic Alzheimer's disease mice revealed by a battery of object-based tests.
- Author
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Creighton SD, Mendell AL, Palmer D, Kalisch BE, MacLusky NJ, Prado VF, Prado MAM, and Winters BD
- Subjects
- Animals, Behavior, Animal, Disease Models, Animal, Female, Male, Mice, Mice, Transgenic, Neuropsychological Tests, Alzheimer Disease pathology, Cognitive Dysfunction pathology
- Abstract
Object recognition tasks detect cognitive deficits in transgenic Alzheimer's disease (AD) mouse models. Object recognition, however, is not a unitary process, and there are many uncharacterized facets of object processing with relevance to AD. We therefore systematically evaluated object processing in 5xFAD and 3xTG AD mice to clarify the nature of object recognition-related deficits. Twelve-month-old male and female 5xFAD and 3xTG mice were assessed on tasks for object identity recognition, spatial recognition, and multisensory object perception. Memory and multisensory perceptual impairments were observed, with interesting dissociations between transgenic AD strains and sex that paralleled neuropathological changes. Overreliance on the widespread "object recognition" task threatens to slow discovery of potentially significant and clinically relevant behavioural effects related to this multifaceted cognitive function. The current results support the use of carefully designed object-based test batteries to clarify the relationship between "object recognition" impairments and specific aspects of AD pathology in rodent models.
- Published
- 2019
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44. Exercise-linked FNDC5/irisin rescues synaptic plasticity and memory defects in Alzheimer's models.
- Author
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Lourenco MV, Frozza RL, de Freitas GB, Zhang H, Kincheski GC, Ribeiro FC, Gonçalves RA, Clarke JR, Beckman D, Staniszewski A, Berman H, Guerra LA, Forny-Germano L, Meier S, Wilcock DM, de Souza JM, Alves-Leon S, Prado VF, Prado MAM, Abisambra JF, Tovar-Moll F, Mattos P, Arancio O, Ferreira ST, and De Felice FG
- Subjects
- Adolescent, Adult, Aged, Alzheimer Disease cerebrospinal fluid, Alzheimer Disease genetics, Animals, Brain metabolism, Brain pathology, Disease Models, Animal, Down-Regulation, Female, Fibronectins cerebrospinal fluid, Fibronectins genetics, Humans, Long-Term Potentiation, Male, Mice, Inbred C57BL, Middle Aged, Neuroprotective Agents pharmacology, Neuroprotective Agents therapeutic use, RNA, Messenger genetics, RNA, Messenger metabolism, Recombinant Proteins pharmacology, Recombinant Proteins therapeutic use, Signal Transduction, Alzheimer Disease metabolism, Alzheimer Disease physiopathology, Fibronectins metabolism, Memory Disorders complications, Memory Disorders physiopathology, Neuronal Plasticity, Physical Conditioning, Animal
- Abstract
Defective brain hormonal signaling has been associated with Alzheimer's disease (AD), a disorder characterized by synapse and memory failure. Irisin is an exercise-induced myokine released on cleavage of the membrane-bound precursor protein fibronectin type III domain-containing protein 5 (FNDC5), also expressed in the hippocampus. Here we show that FNDC5/irisin levels are reduced in AD hippocampi and cerebrospinal fluid, and in experimental AD models. Knockdown of brain FNDC5/irisin impairs long-term potentiation and novel object recognition memory in mice. Conversely, boosting brain levels of FNDC5/irisin rescues synaptic plasticity and memory in AD mouse models. Peripheral overexpression of FNDC5/irisin rescues memory impairment, whereas blockade of either peripheral or brain FNDC5/irisin attenuates the neuroprotective actions of physical exercise on synaptic plasticity and memory in AD mice. By showing that FNDC5/irisin is an important mediator of the beneficial effects of exercise in AD models, our findings place FNDC5/irisin as a novel agent capable of opposing synapse failure and memory impairment in AD.
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- 2019
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45. Fast and slow-twitching muscles are differentially affected by reduced cholinergic transmission in mice deficient for VAChT: A mouse model for congenital myasthenia.
- Author
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Magalhães-Gomes MPS, Motta-Santos D, Schetino LPL, Andrade JN, Bastos CP, Guimarães DAS, Vaughan SK, Martinelli PM, Guatimosim S, Pereira GS, Coimbra CC, Prado VF, Prado MAM, Valdez G, and Guatimosim C
- Subjects
- Acetylcholine antagonists & inhibitors, Animals, Disease Models, Animal, Mice, Transgenic, Muscle, Skeletal drug effects, Myasthenic Syndromes, Congenital genetics, Neuromuscular Junction metabolism, Vesicular Acetylcholine Transport Proteins metabolism, Acetylcholine metabolism, Muscle, Skeletal metabolism, Myasthenic Syndromes, Congenital metabolism, Synaptic Vesicles metabolism
- Abstract
Congenital myasthenic syndromes (CMS) result from reduced cholinergic transmission at neuromuscular junctions (NMJs). While the etiology of CMS varies, the disease is characterized by muscle weakness. To date, it remains unknown if CMS causes long-term and irreversible changes to skeletal muscles. In this study, we examined skeletal muscles in a mouse line with reduced expression of Vesicular Acetylcholine Transporter (VAChT, mouse line herein called VAChT-KD
HOM ). We examined this mouse line for several reasons. First, VAChT plays a central function in loading acetylcholine (ACh) into synaptic vesicles and releasing it at NMJs, in addition to other cholinergic nerve endings. Second, loss of function mutations in VAChT causes myasthenia in humans. Importantly, VAChT-KDHOM present with reduced ACh and muscle weakness, resembling CMS. We evaluated the morphology, fiber type (myosin heavy chain isoforms), and expression of muscle-related genes in the extensor digitorum longus (EDL) and soleus muscles. This analysis revealed that while muscle fibers atrophy in the EDL, they hypertrophy in the soleus muscle of VAChT-KDHOM mice. Along with these cellular changes, skeletal muscles exhibit altered levels of markers for myogenesis (Pax-7, Myogenin, and MyoD), oxidative metabolism (PGC1-α and MTND1), and protein degradation (Atrogin1 and MuRF1) in VAChT-KDHOM mice. Importantly, we demonstrate that deleterious changes in skeletal muscles and motor deficits can be partially reversed following the administration of the cholinesterase inhibitor, pyridostigmine in VAChT-KDHOM mice. These findings reveal that fast and slow type muscles differentially respond to cholinergic deficits. Additionally, this study shows that the adverse effects of cholinergic transmission, as in the case of CMS, on fast and slow type skeletal muscles are reversible., (Copyright © 2018. Published by Elsevier Ltd.)- Published
- 2018
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46. Estradiol effect on short-term object memory under hypocholinergic condition.
- Author
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Pereira LM, Guimarães IM, Oliveira VEM, Bastos CP, Ribeiro FM, Prado VF, Prado MAM, and Pereira GS
- Subjects
- Acetylcholine metabolism, Animals, Estrogens pharmacology, Female, Hippocampus drug effects, Hippocampus metabolism, Male, Memory Disorders metabolism, Memory, Short-Term physiology, Mice, Transgenic, Ovariectomy, RNA, Messenger metabolism, Receptors, Estrogen metabolism, Recognition, Psychology physiology, Sex Factors, Vesicular Acetylcholine Transport Proteins genetics, Estradiol pharmacology, Memory Disorders drug therapy, Memory, Short-Term drug effects, Nootropic Agents pharmacology, Recognition, Psychology drug effects, Vesicular Acetylcholine Transport Proteins deficiency
- Abstract
Estrogens positively affect object recognition memory (ORM). However, whether this effect rely on acetylcholine is unknown. Here we investigated if 17β-estradiol (E2) would be able to recover ORM deficits in animals with decreased expression of the Vesicular Acetylcholine Transporter (VAChT KD
HET ). We found that E2 improved short-term ORM (STM) in VAChT KDHET male and in OVX female mutant mice. However, E2 did not recover long-term (LTM) ORM in both sexes. Next, we tested whether hippocampal ERs activation could also rescue STM in mutant mice. Our results showed that ERα seems to be both sufficient and necessary for STM consolidation in female VAChT KDHET . Differently, in male, both ERα and ERβ activation recovered STM. In addition, we tested whether mRNA level of estrogen receptors (ER) is also sensitive to VAChT expression. Female mutant mice showed lower levels of ER alpha (ERα) mRNA in the hippocampus, while no differences in male were observed. Together, our results showed that under hypocholinergic function, E2 improve short-term object recognition in both male and female. Furthermore, we showed that changes in VAChT expression might potentially modulate hippocampal ERα expression in a sex-dependent-manner., (Copyright © 2018. Published by Elsevier Inc.)- Published
- 2018
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47. Histamine H 3 Receptors Decrease Dopamine Release in the Ventral Striatum by Reducing the Activity of Striatal Cholinergic Interneurons.
- Author
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Varaschin RK, Osterstock G, Ducrot C, Leino S, Bourque MJ, Prado MAM, Prado VF, Salminen O, Rannanpää Née Nuutinen S, and Trudeau LE
- Subjects
- Animals, Dopaminergic Neurons drug effects, Dopaminergic Neurons metabolism, Female, Histamine Agonists pharmacology, Interneurons drug effects, Male, Membrane Potentials drug effects, Membrane Potentials physiology, Methylhistamines pharmacology, Mice, Inbred C57BL, Mice, Transgenic, Optogenetics, RNA, Messenger metabolism, Synaptosomes metabolism, Tissue Culture Techniques, Ventral Striatum drug effects, Acetylcholine metabolism, Dopamine metabolism, Interneurons metabolism, Receptors, Histamine H3 metabolism, Ventral Striatum metabolism
- Abstract
Histamine H
3 receptors are widely distributed Gi -coupled receptors whose activation reduces neuronal activity and inhibits release of numerous neurotransmitters. Although these receptors are abundantly expressed in the striatum, their modulatory role on activity-dependent dopamine release is not well understood. Here, we observed that histamine H3 receptor activation indirectly diminishes dopamine overflow in the ventral striatum by reducing cholinergic interneuron activity. Acute brain slices from C57BL/6 or channelrhodopsin-2-transfected DAT-cre mice were obtained, and dopamine transients evoked either electrically or optogenetically were measured by fast-scan cyclic voltammetry. The H3 agonist α-methylhistamine significantly reduced electrically- evoked dopamine overflow, an effect blocked by the nicotinic acetylcholine receptor antagonist dihydro-β-erythroidine, suggesting involvement of cholinergic interneurons. None of the drug treatments targeting H3 receptors affected optogenetically evoked dopamine overflow, indicating that direct H3 -modulation of dopaminergic axons is unlikely. Next, we used qPCR and confirmed the expression of histamine H3 receptor mRNA in cholinergic interneurons, both in ventral and dorsal striatum. Activation of H3 receptors by α-methylhistamine reduced spontaneous firing of cholinergic interneurons in the ventral, but not in the dorsal striatum. Resting membrane potential and number of spontaneous action potentials in ventral-striatal cholinergic interneurons were significantly reduced by α-methylhistamine. Acetylcholine release from isolated striatal synaptosomes, however, was not altered by α-methylhistamine. Together, these results indicate that histamine H3 receptors are important modulators of dopamine release, specifically in the ventral striatum, and that they do so by decreasing the firing rate of cholinergic neurons and, consequently, reducing cholinergic tone on dopaminergic axons., (Copyright © 2018 IBRO. Published by Elsevier Ltd. All rights reserved.)- Published
- 2018
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48. Mechanisms of neuroprotection against ischemic insult by stress-inducible phosphoprotein-1/prion protein complex.
- Author
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Beraldo FH, Ostapchenko VG, Xu JZ, Di Guglielmo GM, Fan J, Nicholls PJ, Caron MG, Prado VF, and Prado MAM
- Subjects
- Activin Receptors, Type I metabolism, Animals, Apoptosis physiology, Cells, Cultured, Female, Male, Mice, Mice, Inbred C57BL, Protein Binding, Signal Transduction physiology, alpha7 Nicotinic Acetylcholine Receptor metabolism, Brain Ischemia metabolism, Heat-Shock Proteins metabolism, Neuroprotection physiology, Prion Proteins metabolism
- Abstract
Stress-inducible phosphoprotein 1 (STI1) acts as a neuroprotective factor in the ischemic brain and its levels are increased following ischemia. Previous work has suggested that some of these STI1 actions in a stroke model depend on the recruitment of bone marrow-derived stem cells to improve outcomes after ischemic insult. However, STI1 can directly increase neuroprotective signaling in neurons by engaging with the cellular prion protein (PrP
C ) and activating α7 nicotinic acetylcholine receptors (α7nAChR). Given that α7nAChR activation has also been involved in neuroprotection in stroke, it is possible that STI1 can have direct actions on neurons to prevent deleterious consequences of ischemic insults. Here, we tested this hypothesis by exposing primary neuronal cultures to 1-h oxygen-glucose deprivation (OGD) and reperfusion and assessing signaling pathways activated by STI1/PrPC . Our results demonstrated that STI1 treatment significantly decreased apoptosis and cell death in mouse neurons submitted to OGD in a manner that was dependent on PrPC and α7nAChR, but also on the activin A receptor 1 (ALK2), which has emerged as a signaling partner of STI1. Interestingly, pharmacological inhibition of the ALK2 receptor prevented neuroprotection by STI1, while activation of ALK2 receptors by bone morphogenetic protein 4 (BMP4) either before or after OGD was effective in decreasing neuronal death induced by ischemia. We conclude that PrPC /STI1 engagement and its subsequent downstream signaling cascades involving α7nAChR as well as the ALK2 receptor may be activated in neurons by increased levels of STI1. This signaling pathway protects neurons from ischemic insults., (© 2017 International Society for Neurochemistry.)- Published
- 2018
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49. Vesicular acetylcholine transporter (VAChT) over-expression induces major modifications of striatal cholinergic interneuron morphology and function.
- Author
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Janickova H, Prado VF, Prado MAM, El Mestikawy S, and Bernard V
- Abstract
Striatal cholinergic interneurons (CIN) are pivotal for the regulation of the striatal network. Acetylcholine (ACh) released by CIN is centrally involved in reward behavior as well as locomotor or cognitive functions. Recently, BAC transgenic mice expressing channelrhodopsin-2 (ChR2) protein under the control of the choline acetyltransferase (ChAT) promoter (ChAT-ChR2) and displaying almost 50 extra copies of the VAChT gene were used to dissect cholinergic circuit connectivity and function using optogenetic approaches. These mice display over-expression of the vesicular acetylcholine transporter (VAChT) and increased cholinergic tone. Consequently, ChAT-ChR2 mice are a valuable model to investigate hypercholinergic phenotypes. Previous experiments established that ChAT-ChR2 mice display an increased sensitivity to amphetamine induced-locomotor activity and stereotypes. In the present report, we analyzed the impact of VAChT over-expression in the striatum of ChAT-ChR2 mice. ChAT-ChR2 mice displayed increased locomotor sensitization in response to low dose of cocaine. In addition, we observed a dramatic remodeling of the morphology of CIN in ChAT-ChR2 transgenic mice. VAChT immunolabeling was markedly enhanced in the soma and terminal of CIN from ChAT-ChR2 mice as previously shown (Crittenden et al. 2014). Interestingly, the number of cholinergic varicosities was markedly reduced (-87%) whereas their size was significantly increased (+177%). Moreover, VAChT over-expression dramatically modified its trafficking along the somatodendritic and axonal arbor. These findings demonstrate that ChAT-ChR2 mice present major alterations of CIN neuronal morphology and increased behavioral sensitization to cocaine, supporting the notion that the increased levels of VAChT observed in these mice make them fundamentally different from wild-type mice., (© 2017 International Society for Neurochemistry.)
- Published
- 2017
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50. Cholinergic/glutamatergic co-transmission in striatal cholinergic interneurons: new mechanisms regulating striatal computation.
- Author
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Kljakic O, Janickova H, Prado VF, and Prado MAM
- Subjects
- Animals, Corpus Striatum drug effects, Humans, Interneurons drug effects, Neostriatum drug effects, Synaptic Transmission physiology, Acetylcholine metabolism, Cholinergic Agents pharmacology, Corpus Striatum metabolism, Interneurons metabolism, Neostriatum metabolism, Synaptic Transmission drug effects
- Abstract
It is well established that neurons secrete neuropeptides and ATP with classical neurotransmitters; however, certain neuronal populations are also capable of releasing two classical neurotransmitters by a process named co-transmission. Although there has been progress in our understanding of the molecular mechanism underlying co-transmission, the individual regulation of neurotransmitter secretion and the functional significance of this neuronal 'bilingualism' is still unknown. Striatal cholinergic interneurons (CINs) have been shown to secrete glutamate (Glu) in addition to acetylcholine (ACh) and are recognized for their role in the regulation of striatal circuits and behavior. Our review highlights the recent research into identifying mechanisms that regulate the secretion and function of Glu and ACh released by CINs and the roles these neurons play in regulating dopamine secretion and striatal activity. In particular, we focus on how the transporters for ACh (VAChT) and Glu (VGLUT3) influence the storage of neurotransmitters in CINs. We further discuss how these individual neurotransmitters regulate striatal computation and distinct aspects of behavior that are regulated by the striatum. We suggest that understanding the distinct and complementary functional roles of these two neurotransmitters may prove beneficial in the development of therapies for Parkinson's disease and addiction. Overall, understanding how Glu and ACh secreted by CINs impacts striatal activity may provide insight into how different populations of 'bilingual' neurons are able to develop sophisticated regulation of their targets by interacting with multiple receptors but also by regulating each other's vesicular storage. This is an article for the special issue XVth International Symposium on Cholinergic Mechanisms., (© 2017 International Society for Neurochemistry.)
- Published
- 2017
- Full Text
- View/download PDF
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