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1.
Proteomics ; 22(11-12): e2100244, 2022 06.
Article in English | MEDLINE | ID: mdl-35355420

ABSTRACT

A major challenge in managing depression is that antidepressant drugs take a long time to exert their therapeutic effects. For the development of faster-acting treatments, it is crucial to get an improved understanding of the molecular mechanisms underlying antidepressant mode of action. Here, we used a novel mass spectrometry-based workflow to investigate how antidepressant treatment affects brain protein turnover at single-cell and subcellular resolution. We combined stable isotope metabolic labeling, quantitative Tandem Mass Spectrometry (qTMS) and Multi-isotope Imaging Mass Spectrometry (MIMS) to simultaneously quantify and image protein synthesis and turnover in hippocampi of mice treated with the antidepressant paroxetine. We identified changes in turnover of individual hippocampal proteins that reveal altered metabolism-mitochondrial processes and report on subregion-specific turnover patterns upon paroxetine treatment. This workflow can be used to investigate brain protein turnover changes in vivo upon pharmacological interventions at a resolution and precision that has not been possible with other methods to date. Our results reveal acute paroxetine effects on brain protein turnover and shed light on antidepressant mode of action.


Subject(s)
Antidepressive Agents , Paroxetine , Animals , Antidepressive Agents/metabolism , Antidepressive Agents/pharmacology , Antidepressive Agents/therapeutic use , Hippocampus/metabolism , Isotope Labeling/methods , Isotopes/metabolism , Isotopes/pharmacology , Mice , Paroxetine/metabolism , Paroxetine/pharmacology , Tandem Mass Spectrometry
2.
Eur J Neurosci ; 53(9): 3002-3018, 2021 05.
Article in English | MEDLINE | ID: mdl-33226682

ABSTRACT

Dysfunction of metabolic pathways characterises a plethora of common pathologies and has emerged as an underlying hallmark of disease phenotypes. Here, we focus on psychiatric disorders and brain tumours and explore changes in the interplay between glycolysis and mitochondrial energy metabolism in the brain. We discuss alterations in glycolysis versus core mitochondrial metabolic pathways, such as the tricarboxylic acid cycle and oxidative phosphorylation, in major psychiatric disorders and brain tumours. We investigate potential common patterns of altered mitochondrial metabolism in different brain regions and sample types and explore how changes in mitochondrial number, shape and morphology affect disease-related manifestations. We also highlight the potential of pharmacologically targeting mitochondria to achieve therapeutic effects.


Subject(s)
Brain Neoplasms , Mental Disorders , Energy Metabolism , Glycolysis , Humans , Mitochondria/metabolism , Oxidative Phosphorylation
3.
Stress ; 24(6): 952-964, 2021 11.
Article in English | MEDLINE | ID: mdl-34553679

ABSTRACT

Psychological stress and stress-related disorders constitute a major health problem in modern societies. Although the brain circuits involved in emotional processing are intensively studied, little is known about the implication of cerebellum in stress responses whereas the molecular changes induced by stress exposure in cerebellum remain largely unexplored. Here, we investigated the effects of acute stress exposure on mouse cerebellum. We used a forced swim test (FST) paradigm as an acute stressor. We then analyzed the cerebellar metabolomic profiles of stressed (n = 11) versus control (n = 11) male CD1 mice by a Nuclear Magnetic Resonance (NMR)-based, untargeted metabolomics approach. Our results showed altered levels of 19 out of the 47 annotated metabolites, which are implicated in neurotransmission and N-acetylaspartic acid (NAA) turnover, as well as in energy and purine/pyrimidine metabolism. We also correlated individual metabolite levels with FST behavioral parameters, and reported associations between FST readouts and levels of 4 metabolites. This work indicates an altered metabolomic signature after acute stress in the cerebellum and highlights a previously unexplored involvement of cerebellum in stress responses.


Subject(s)
Metabolomics , Stress, Psychological , Animals , Cerebellum/metabolism , Disease Models, Animal , Male , Metabolomics/methods , Mice , Stress, Psychological/metabolism , Swimming
4.
Cereb Cortex ; 27(4): 2580-2591, 2017 04 01.
Article in English | MEDLINE | ID: mdl-27073221

ABSTRACT

Tau protein in dendrites and synapses has been recently implicated in synaptic degeneration and neuronal malfunction. Chronic stress, a well-known inducer of neuronal/synaptic atrophy, triggers hyperphosphorylation of Tau protein and cognitive deficits. However, the cause-effect relationship between these events remains to be established. To test the involvement of Tau in stress-induced impairments of cognition, we investigated the impact of stress on cognitive behavior, neuronal structure, and the synaptic proteome in the prefrontal cortex (PFC) of Tau knock-out (Tau-KO) and wild-type (WT) mice. Whereas exposure to chronic stress resulted in atrophy of apical dendrites and spine loss in PFC neurons as well as significant impairments in working memory in WT mice, such changes were absent in Tau-KO animals. Quantitative proteomic analysis of PFC synaptosomal fractions, combined with transmission electron microscopy analysis, suggested a prominent role for mitochondria in the regulation of the effects of stress. Specifically, chronically stressed animals exhibit Tau-dependent alterations in the levels of proteins involved in mitochondrial transport and oxidative phosphorylation as well as in the synaptic localization of mitochondria in PFC. These findings provide evidence for a causal role of Tau in mediating stress-elicited neuronal atrophy and cognitive impairment and indicate that Tau may exert its effects through synaptic mitochondria.


Subject(s)
Mitochondria/pathology , Prefrontal Cortex/pathology , Stress, Psychological/complications , Synapses/pathology , tau Proteins/metabolism , Animals , Atrophy , Chromatography, High Pressure Liquid , Dendrites/pathology , Dendrites/ultrastructure , Disease Models, Animal , Male , Mass Spectrometry , Mice , Mice, Inbred C57BL , Mice, Knockout , Microscopy, Electron, Transmission , Proteomics
5.
Neurogenetics ; 15(3): 201-12, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24928144

ABSTRACT

'Neuroinflammation' has become a widely applied term in the basic and clinical neurosciences but there is no generally accepted neuropathological tissue correlate. Inflammation, which is characterized by the presence of perivascular infiltrates of cells of the adaptive immune system, is indeed seen in the central nervous system (CNS) under certain conditions. Authors who refer to microglial activation as neuroinflammation confuse this issue because autoimmune neuroinflammation serves as a synonym for multiple sclerosis, the prototypical inflammatory disease of the CNS. We have asked the question whether a data-driven, unbiased in silico approach may help to clarify the nomenclatorial confusion. Specifically, we have examined whether unsupervised analysis of microarray data obtained from human cerebral cortex of Alzheimer's, Parkinson's and schizophrenia patients would reveal a degree of relatedness between these diseases and recognized inflammatory conditions including multiple sclerosis. Our results using two different data analysis methods provide strong evidence against this hypothesis demonstrating that very different sets of genes are involved. Consequently, the designations inflammation and neuroinflammation are not interchangeable. They represent different categories not only at the histophenotypic but also at the transcriptomic level. Therefore, non-autoimmune neuroinflammation remains a term in need of definition.


Subject(s)
Alzheimer Disease/genetics , Encephalitis/genetics , Multiple Sclerosis/genetics , Parkinson Disease/genetics , Schizophrenia/genetics , Transcriptome , Cluster Analysis , Computational Biology , Computer Simulation , Gene Expression Profiling , Humans , Immunoglobulins/metabolism , Inflammation/genetics , Intercellular Signaling Peptides and Proteins/metabolism
6.
Neurosci Biobehav Rev ; : 105837, 2024 Jul 30.
Article in English | MEDLINE | ID: mdl-39089419

ABSTRACT

Elucidating the molecular mechanisms of psychopathology is crucial for optimized diagnosis and treatment. Accumulating literature has underlined how mitochondrial bioenergetics affect major psychiatric disorders. However, how mitochondrial dynamics, a term addressing mitochondria quality control, including mitochondrial fission, fusion, biogenesis and mitophagy, is implicated in psychopathologies remains elusive. In this review we summarize the existing literature on mitochondrial dynamics perturbations in psychiatric disorders/neuropsychiatric phenotypes. We include preclinical/clinical literature on mitochondrial dynamics recalibrations in anxiety, depression, post-traumatic stress disorder (PTSD), bipolar disorder and schizophrenia. We discuss alterations in mitochondrial network, morphology and shape; molecular markers of the mitochondrial dynamics machinery and mitochondrial DNA copy number (mtDNAcn) in animal models and human cohorts in brain and peripheral material. By looking for common altered mitochondrial dynamics patterns across diagnoses/phenotypes, we highlight mitophagy and biogenesis as regulators of anxiety and depression pathophysiology, respectively, as well as the fusion mediator dynamin-like 120kDa protein (Opa1) as a molecular hub contributing to psychopathology. Finally, we comment on limitations and future directions in this novel neuropsychiatry field.

7.
Curr Neuropharmacol ; 22(5): 884-903, 2024.
Article in English | MEDLINE | ID: mdl-37448366

ABSTRACT

Despite intensive research efforts to understand the molecular underpinnings of psychological stress and stress responses, the underlying molecular mechanisms remain largely elusive. Towards this direction, a plethora of stress rodent models have been established to investigate the effects of exposure to different stressors. To decipher affected molecular pathways in a holistic manner in these models, metabolomics approaches addressing altered, small molecule signatures upon stress exposure in a high-throughput, quantitative manner provide insightful information on stress-induced systemic changes in the brain. In this review, we discuss stress models in mice and rats, followed by mass spectrometry (MS) and nuclear magnetic resonance (NMR) metabolomics studies. We particularly focus on acute, chronic and early life stress paradigms, highlight how stress is assessed at the behavioral and molecular levels and focus on metabolomic outcomes in the brain and peripheral material such as plasma and serum. We then comment on common metabolomics patterns across different stress models and underline the need for unbiased -omics methodologies and follow-up studies of metabolomics outcomes to disentangle the complex pathobiology of stress and pertinent psychopathologies.


Subject(s)
Metabolome , Rodentia , Rats , Mice , Animals , Metabolomics/methods , Brain , Stress, Psychological
8.
Mol Neurobiol ; 2024 May 18.
Article in English | MEDLINE | ID: mdl-38761326

ABSTRACT

Early handling (EH), the brief separation of pups from their mother during early life, has been shown to exert beneficial effects. However, the impact of EH in a high anxiety background as well as the role of brain mitochondria in shaping EH-driven responses remain elusive.Here, we used a high (HAB) vs. normal (NAB) anxiety-related behavior mouse model to study how EH affects pup and dam behavior in divergent anxiety backgrounds. We also investigated EH-induced effects at the protein and mRNA levels in adult male HAB mice in the hypothalamus, the prefrontal cortex, and the hippocampus by examining the same mitochondrial/energy pathways and mitochondrial dynamics mechanisms (fission, fusion, biogenesis, and mitophagy) in all three brain regions.EH exerts anxiolytic effects in adult HAB but not NAB male mice and does not affect HAB or NAB maternal behavior, although basal HAB vs. NAB maternal behaviors differ. In adult HAB male mice, EH does not impact oxidative phosphorylation (OXPHOS) and oxidative stress in any of the brain regions studied but leads to increased protein expression of glycolysis enzymes and a correlation of anxiety-related behavior with Krebs cycle enzymes in HAB mice in the hypothalamus. Intriguingly, EH alters mitochondrial dynamics by increasing hypothalamic DRP1, OPA1, and PGC1a protein levels. At the mRNA level, we observe altered, EH-driven mitochondrial dynamics mRNA signatures which predominantly affect the prefrontal cortex.Taken together, our results show that EH exerts anxiolytic effects in adulthood in high anxiety and modulates mitochondrial dynamics pathways in a brain region-specific manner.

9.
Neuropharmacology ; 248: 109870, 2024 May 01.
Article in English | MEDLINE | ID: mdl-38401791

ABSTRACT

Delayed therapeutic responses and limited efficacy are the main challenges of existing antidepressant drugs, thereby incentivizing the search for new potential treatments. Cannabidiol (CBD), non-psychotomimetic component of cannabis, has shown promising antidepressant effects in different rodent models, but its mechanism of action remains unclear. Herein, we investigated the antidepressant-like effects of repeated CBD treatment on behavior, neuroplasticity markers and lipidomic profile in the prefrontal cortex (PFC) of Flinders Sensitive Line (FSL), a genetic animal model of depression, and their control counterparts Flinders Resistant Line (FRL) rats. Male FSL animals were treated with CBD (10 mg/kg; i.p.) or vehicle (7 days) followed by Open Field Test (OFT) and the Forced Swimming Test (FST). The PFC was analyzed by a) western blotting to assess markers of synaptic plasticity and cannabinoid signaling in synaptosome and cytosolic fractions; b) mass spectrometry-based lipidomics to investigate endocannabinoid levels (eCB). CBD attenuated the increased immobility observed in FSL, compared to FRL in FST, without changing the locomotor behavior in the OFT. In synaptosomes, CBD increased ERK1, mGluR5, and Synaptophysin, but failed to reverse the reduced CB1 and CB2 levels in FSL rats. In the cytosolic fraction, CBD increased ERK2 and decreased mGluR5 expression in FSL rats. Surprisingly, there were no significant changes in eCB levels in response to CBD treatment. These findings suggest that CBD effects in FSL animals are associated with changes in synaptic plasticity markers involving mGluR5, ERK1, ERK2, and synaptophysin signaling in the PFC, without increasing the levels of endocannabinoids in this brain region.


Subject(s)
Cannabidiol , Depression , Rats , Male , Animals , Depression/drug therapy , Depression/genetics , Cannabidiol/pharmacology , Endocannabinoids/metabolism , Synaptophysin/metabolism , Antidepressive Agents/pharmacology , Prefrontal Cortex , Neuronal Plasticity , Disease Models, Animal
10.
Brain Sci ; 14(2)2024 Jan 28.
Article in English | MEDLINE | ID: mdl-38391714

ABSTRACT

Developmental dyslexia (DD) is a learning disorder. Although risk genes have been identified, environmental factors, and particularly stress arising from constant difficulties, have been associated with the occurrence of DD by affecting brain plasticity and function, especially during critical neurodevelopmental stages. In this work, electroencephalogram (EEG) findings were coupled with the genetic and epigenetic molecular signatures of individuals with DD and matched controls. Specifically, we investigated the genetic and epigenetic correlates of key stress-associated genes (NR3C1, NR3C2, FKBP5, GILZ, SLC6A4) with psychological characteristics (depression, anxiety, and stress) often included in DD diagnostic criteria, as well as with brain EEG findings. We paired the observed brain rhythms with the expression levels of stress-related genes, investigated the epigenetic profile of the stress regulator glucocorticoid receptor (GR) and correlated such indices with demographic findings. This study presents a new interdisciplinary approach and findings that support the idea that stress, attributed to the demands of the school environment, may act as a contributing factor in the occurrence of the DD phenotype.

11.
Mol Cell Proteomics ; 10(12): M111.008110, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21862759

ABSTRACT

Depression and anxiety disorders affect a great number of people worldwide. Whereas singular factors have been associated with the pathogenesis of psychiatric disorders, growing evidence emphasizes the significance of dysfunctional neural circuits and signaling pathways. Hence, a systems biology approach is required to get a better understanding of psychiatric phenotypes such as depression and anxiety. Furthermore, the availability of biomarkers for these disorders is critical for improved diagnosis and monitoring treatment response. In the present study, a mouse model presenting with robust high versus low anxiety phenotypes was subjected to thorough molecular biomarker and pathway discovery analyses. Reference animals were metabolically labeled with the stable (15)N isotope allowing an accurate comparison of protein expression levels between the high anxiety-related behavior versus low anxiety-related behavior mouse lines using quantitative mass spectrometry. Plasma metabolomic analyses identified a number of small molecule biomarkers characteristic for the anxiety phenotype with particular focus on myo-inositol and glutamate as well as the intermediates involved in the tricarboxylic acid cycle. In silico analyses suggested pathways and subnetworks as relevant for the anxiety phenotype. Our data demonstrate that the high anxiety-related behavior and low anxiety-related behavior mouse model is a valuable tool for anxiety disorder drug discovery efforts.


Subject(s)
Anxiety Disorders/blood , Metabolic Networks and Pathways , Amino Acid Sequence , Animals , Anxiety Disorders/genetics , Biomarkers/blood , Carbonic Anhydrase II/blood , Carbonic Anhydrase II/chemistry , Glutamic Acid/blood , Hippocampus/enzymology , Inositol/blood , Lactoylglutathione Lyase/chemistry , Lactoylglutathione Lyase/metabolism , Male , Metabolomics , Molecular Sequence Data , Multifactorial Inheritance , Peptide Fragments/chemistry , Prealbumin/chemistry , Prealbumin/metabolism , Protein Array Analysis , Proteomics , Serum Amyloid P-Component/chemistry , Serum Amyloid P-Component/metabolism
12.
Proteomics ; 12(4-5): 736-47, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22247077

ABSTRACT

Proteomics has provided researchers with a sophisticated toolbox of labeling-based and label-free quantitative methods. These are now being applied in neuroscience research where they have already contributed to the elucidation of fundamental mechanisms and the discovery of candidate biomarkers. In this review, we evaluate and compare labeling-based and label-free quantitative proteomic techniques for applications in neuroscience research. We discuss the considerations required for the analysis of brain and central nervous system specimens, the experimental design of quantitative proteomic workflows as well as the feasibility, advantages, and disadvantages of the available techniques for neuroscience-oriented questions. Furthermore, we assess the use of labeled standards as internal controls for comparative studies in humans and review applications of labeling-based and label-free mass spectrometry approaches in relevant model organisms and human subjects. Providing a comprehensive guide of feasible and meaningful quantitative proteomic methodologies for neuroscience research is crucial not only for overcoming current limitations but also for gaining useful insights into brain function and translating proteomics from bench to bedside.


Subject(s)
Brain Chemistry , Neurosciences/methods , Proteome/analysis , Proteomics/methods , Spinal Cord/chemistry , Animals , Biomarkers/analysis , Brain/cytology , Brain/metabolism , Humans , Mass Spectrometry/methods , Spinal Cord/cytology , Spinal Cord/metabolism , Staining and Labeling
13.
Proteomics ; 12(21): 3121-8, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22887715

ABSTRACT

Several techniques based on stable isotope labeling are used for quantitative MS. These include stable isotope metabolic labeling methods for cells in culture as well as live organisms with the assumption that the stable isotope has no effect on the proteome. Here, we investigate the (15) N isotope effect on Escherichia coli cultures that were grown in either unlabeled ((14) N) or (15) N-labeled media by LC-ESI-MS/MS-based relative protein quantification. Consistent protein expression level differences and altered growth rates were observed between (14) N and (15) N-labeled cultures. Furthermore, targeted metabolite analyses revealed altered metabolite levels between (14) N and (15) N-labeled bacteria. Our data demonstrate for the first time that the introduction of the (15) N isotope affects protein and metabolite levels in E. coli and underline the importance of implementing controls for unbiased protein quantification using stable isotope labeling techniques.


Subject(s)
Escherichia coli/metabolism , Isotope Labeling/methods , Nitrogen Isotopes/chemistry , Proteomics/methods , Chromatography, Liquid , Neutrons , Proteome/analysis , Proteome/chemistry , Proteome/metabolism , Tandem Mass Spectrometry
14.
Proteomics ; 12(15-16): 2421-7, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22700377

ABSTRACT

Stable isotope labeling techniques hold great potential for accurate quantitative proteomics comparisons by MS. To investigate the effect of stable isotopes in vivo, we metabolically labeled high anxiety-related behavior (HAB) mice with the heavy nitrogen isotope (15)N. (15)N-labeled HAB mice exhibited behavioral alterations compared to unlabeled ((14)N) HAB mice in their depression-like phenotype. To correlate behavioral alterations with changes on the molecular level, we explored the (15)N isotope effect on the brain proteome by comparing protein expression levels between (15)N-labeled and (14)N HAB mouse brains using quantitative MS. By implementing two complementary in silico pathway analysis approaches, we were able to identify altered networks in (15)N-labeled HAB mice, including major metabolic pathways such as the tricarboxylic acid (TCA) cycle and oxidative phosphorylation. Here, we discuss the affected pathways with regard to their relevance for the behavioral phenotype and critically assess the utility of exploiting the (15)N isotope effect for correlating phenotypic and molecular alterations.


Subject(s)
Anxiety/metabolism , Anxiety/pathology , Isotope Labeling/methods , Signal Transduction , Animals , Behavior, Animal , Disease Models, Animal , Male , Mice , Nitrogen Isotopes , Phenotype , Proteome/metabolism , Proteomics
15.
J Pers Med ; 12(2)2022 Feb 09.
Article in English | MEDLINE | ID: mdl-35207732

ABSTRACT

The primate-specific G72/G30 gene locus has been associated with major psychiatric disorders, such as schizophrenia and bipolar disorder. We have previously generated transgenic mice which carry the G72/G30 locus and express the longest G72 splice variant (LG72) protein encoded by this locus with schizophrenia-related symptoms. Here, we used a multi-omics approach, including quantitative proteomics and metabolomics to investigate molecular alterations in the hippocampus of G72/G30 transgenic (G72Tg) mice. Our proteomics analysis revealed decreased expression of myelin-related proteins and NAD-dependent protein deacetylase sirtuin-2 (Sirt2) as well as increased expression of the scaffolding presynaptic proteins bassoon (Bsn) and piccolo (Pclo) and the cytoskeletal protein plectin (Plec1) in G72Tg compared to wild-type (WT) mice. Metabolomics analysis indicated decreased levels of nicotinate in G72Tg compared to WT hippocampi. Decreased hippocampal protein expression for selected proteins, namely myelin oligodentrocyte glycoprotein (Mog), Cldn11 and myelin proteolipid protein (Plp), was confirmed with Western blot in a larger population of G72Tg and WT mice. The identified molecular pathway alterations shed light on the hippocampal function of LG72 protein in the context of neuropsychiatric phenotypes.

16.
Metabolites ; 11(2)2021 Feb 23.
Article in English | MEDLINE | ID: mdl-33672326

ABSTRACT

Mouse models are widely used to study behavioral phenotypes related to neuropsychiatric disorders. However, different mouse strains vary in their inherent behavioral and molecular characteristics, which needs to be taken into account depending on the nature of the study. Here, we performed a detailed behavioral and molecular comparison of C57BL/6 (B6) and DBA/2 (DBA) mice, two inbred strains commonly used in neuropsychiatric research. We analyzed anxiety-related and depression-like traits, quantified hippocampal and plasma metabolite profiles, and assessed total antioxidant capacity (ΤAC). B6 mice exhibit increased depression-like and decreased anxiety-related behavior compared to DBA mice. Metabolite level differences indicate alterations in amino acid, nucleotide and mitochondrial metabolism that are accompanied by a decreased TAC in B6 compared to DBA mice. Our data reveal multiple behavioral and molecular differences between B6 and DBA mouse strains, which should be considered in the experimental design for phenotype, pharmacological and mechanistic studies relevant for neuropsychiatric disorders.

17.
Electrophoresis ; 31(8): 1294-301, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20309889

ABSTRACT

Synapses play important roles in neurotransmission and neuroplasticity. For an in-depth analysis of the synaptic proteome and phosphoproteome, synaptosomal proteins from whole mouse brain were analyzed by IEF and MS resulting in the largest synaptosome proteome described to date, with 2980 unique proteins identified with two or more peptides. At the same time, 118 synaptosomal phosphoproteins were identified, eight of which are reported for the first time as phosphorylated. Expression of selected proteins in synaptosomes was investigated by Western blot. We demonstrate that IEF is a powerful method to interrogate complex samples such as brain tissue both at the proteome and the phosphoproteome level without the need of additional enrichment for phosphoproteins. The detailed synaptoproteome data set reported here will help to elucidate the molecular complexity of the synapse and contribute to our understanding of synaptic systems biology in health and disease.


Subject(s)
Isoelectric Focusing/methods , Mass Spectrometry/methods , Nerve Tissue Proteins/chemistry , Phosphoproteins/metabolism , Proteome/chemistry , Synaptosomes/chemistry , Amino Acid Sequence , Animals , Blotting, Western , Isoelectric Point , Mice , Molecular Sequence Data , Molecular Weight , Nerve Tissue Proteins/metabolism , Phosphoproteins/chemistry , Phosphoproteins/classification , Proteome/metabolism , Proteomics/methods , Synaptosomes/metabolism
18.
Chemosphere ; 252: 126417, 2020 Aug.
Article in English | MEDLINE | ID: mdl-32200177

ABSTRACT

Cannabinol (CBN) is a degradation product of the cannabis metabolite Δ9-tetrahydrocannabinol. The CBN concentration in cannabis leaves ranges between 0.1 and 1.6% (w/w of dry weight); it increases as the plant ages and its formation is affected by the storage conditions. As CBN has not been extensively studied so far, the need to examine its impact in vivo is imperative due to the increasing use of cannabis globally. In the study herein, the CBN toxicity, effects on heart physiology, morphological malformations, behavioral changes and alterations in metabolic pathways of zebrafish larvae upon CBN exposure to sublethal concentrations were examined. The LD50 value was estimated at 1.12 mg/l. At the same time, malformations in zebrafish larvae increased significantly in a dose-dependent manner and exposure to CBN concentrations greater than 0.75 mg/l provoked abnormalities like pericardial edema, yolk sac anomalies and tail bending. Concentrations above this threshold resulted in elongated and shorter in width hearts and in separation of ventricle from atrium. The total movement distance and velocity were increased in dark and decreased in light conditions, in a concentration-dependent manner. Our results showed that CBN acts both as a stimulant and a sedative, with larvae to exhibit altered velocity and bradycardia, respectively. The metabolomic analysis revealed alterations mainly to amino acids, which are related to acute toxicity and hint towards systemic metabolic and neuropathophysiological changes. Taken together, our data indicate increased toxic effects as CBN exposure concentration increases, which should be taken into consideration when studying the impact of cannabis on organisms.


Subject(s)
Cannabinol/toxicity , Embryo, Nonmammalian/drug effects , Water Pollutants, Chemical/toxicity , Analgesics , Animals , Cannabinoids/toxicity , Cannabis , Dronabinol/analysis , Larva/drug effects , Lethal Dose 50 , Zebrafish/embryology
19.
Proteomics ; 9(17): 4265-70, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19722194

ABSTRACT

Quantitative proteomics using stable isotope labeling strategies combined with MS is an important tool for biomarker discovery. Methods involving stable isotope metabolic labeling result in optimal quantitative accuracy, since they allow the immediate combination of two or more samples. Unfortunately, stable isotope incorporation rates in metabolic labeling experiments using mammalian organisms usually do not reach 100%. As a consequence, protein identifications in (15)N database searches have poor success rates. We report on a strategy that significantly improves the number of (15)N-labeled protein identifications and results in a more comprehensive and accurate relative peptide quantification workflow.


Subject(s)
Databases, Protein , Isotope Labeling/methods , Mass Spectrometry/methods , Proteins/analysis , Amino Acid Sequence , Animals , Mice , Molecular Sequence Data , Nitrogen Isotopes , Peptides/analysis , Peptides/chemistry , Proteins/chemistry
20.
Trends Neurosci ; 42(9): 573-588, 2019 09.
Article in English | MEDLINE | ID: mdl-31362874

ABSTRACT

Accumulating data highlight the contribution of brain mitochondria and bioenergetics to psychiatric disorders and stress-related pathologies. Although anxiety has not received much attention in this booming literature, a bidirectional interplay between anxiety and brain mitochondria and metabolism has recently started to emerge. Substantial observations indicate alterations in mitochondria and metabolism in highly anxious individuals and, conversely, anxiety symptoms in humans suffering from mitochondrial disorders. Genetic and pharmacological efforts have made substantial progress at advancing the causal involvement of specific mitochondrial and metabolic factors in anxiety. In this review, we discuss this converging evidence and highlight the relevance of developing a research focused on targeting mitochondria as an approach to alleviate anxiety.


Subject(s)
Anxiety Disorders/metabolism , Brain/metabolism , Energy Metabolism/physiology , Mitochondrial Diseases/metabolism , Animals , Anxiety Disorders/physiopathology , Brain/physiopathology , Humans , Mitochondria/metabolism , Mitochondrial Diseases/physiopathology , Oxidative Stress/physiology
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