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Ligand-conjugated quantum dots for fast sub-diffraction protein tracking in acute brain slices.
Thal LB, Mann VR, Sprinzen D, McBride JR, Reid KR, Tomlinson ID, McMahon DG, Cohen BE, Rosenthal SJ
(2020) Biomater Sci 8: 837-845
MeSH Terms: Animals, Brain, Brain Chemistry, Ligands, Mice, Microscopy, Fluorescence, Microtomy, Proteins, Quantum Dots, Selenium Compounds, Staining and Labeling, Zinc Compounds
Show Abstract · Added March 18, 2020
Semiconductor quantum dots (QDs) have demonstrated utility in long-term single particle tracking of membrane proteins in live cells in culture. To extend the superior optical properties of QDs to more physiologically relevant cell platforms, such as acute brain slices, we examine the photophysics of compact ligand-conjugated CdSe/CdS QDs using both ensemble and single particle analysis in brain tissue media. We find that symmetric core passivation is critical for both photostability in oxygenated media and for prolonged single particle imaging in brain slices. We then demonstrate the utility of these QDs by imaging single dopamine transporters in acute brain slices, achieving 20 nm localization precision at 10 Hz frame rates. These findings detail design requirements needed for new QD probes in complex living environments, and open the door to physiologically relevant studies that capture the utility of QD probes in acute brain slices.
0 Communities
2 Members
0 Resources
12 MeSH Terms
A gene co-expression network-based analysis of multiple brain tissues reveals novel genes and molecular pathways underlying major depression.
Gerring ZF, Gamazon ER, Derks EM, Major Depressive Disorder Working Group of the Psychiatric Genomics Consortium
(2019) PLoS Genet 15: e1008245
MeSH Terms: Brain Chemistry, Complement C4a, Depressive Disorder, Major, Gene Expression Profiling, Gene Expression Regulation, Gene Regulatory Networks, Genome-Wide Association Study, Humans, Organ Specificity, Quantitative Trait Loci, Sequence Analysis, RNA
Show Abstract · Added July 17, 2019
Major depression is a common and severe psychiatric disorder with a highly polygenic genetic architecture. Genome-wide association studies have successfully identified multiple independent genetic loci that harbour variants associated with major depression, but the exact causal genes and biological mechanisms are largely unknown. Tissue-specific network approaches may identify molecular mechanisms underlying major depression and provide a biological substrate for integrative analyses. We provide a framework for the identification of individual risk genes and gene co-expression networks using genome-wide association summary statistics and gene expression information across multiple human brain tissues and whole blood. We developed a novel gene-based method called eMAGMA that leverages tissue-specific eQTL information to identify 99 biologically plausible risk genes associated with major depression, of which 58 are novel. Among these novel associations is Complement Factor 4A (C4A), recently implicated in schizophrenia through its role in synaptic pruning during postnatal development. Major depression risk genes were enriched in gene co-expression modules in multiple brain tissues and the implicated gene modules contained genes involved in synaptic signalling, neuronal development, and cell transport pathways. Modules enriched with major depression signals were strongly preserved across brain tissues, but were weakly preserved in whole blood, highlighting the importance of using disease-relevant tissues in genetic studies of psychiatric traits. We identified tissue-specific genes and gene co-expression networks associated with major depression. Our novel analytical framework can be used to gain fundamental insights into the functioning of the nervous system in major depression and other brain-related traits.
0 Communities
1 Members
0 Resources
11 MeSH Terms
Iron, Myelin, and the Brain: Neuroimaging Meets Neurobiology.
Möller HE, Bossoni L, Connor JR, Crichton RR, Does MD, Ward RJ, Zecca L, Zucca FA, Ronen I
(2019) Trends Neurosci 42: 384-401
MeSH Terms: Brain, Brain Chemistry, Humans, Iron, Myelin Sheath, Neurobiology, Neuroimaging
Show Abstract · Added March 5, 2020
Although iron is crucial for neuronal functioning, many aspects of cerebral iron biology await clarification. The ability to quantify specific iron forms in the living brain would open new avenues for diagnosis, therapeutic monitoring, and understanding pathogenesis of diseases. A modality that allows assessment of brain tissue composition in vivo, in particular of iron deposits or myelin content on a submillimeter spatial scale, is magnetic resonance imaging (MRI). Multimodal strategies combining MRI with complementary analytical techniques ex vivo have emerged, which may lead to improved specificity. Interdisciplinary collaborations will be key to advance beyond simple correlative analyses in the biological interpretation of MRI data and to gain deeper insights into key factors leading to iron accumulation and/or redistribution associated with neurodegeneration.
Copyright © 2019. Published by Elsevier Ltd.
0 Communities
1 Members
0 Resources
7 MeSH Terms
Imaging Mass Spectrometry: A Perspective.
Caprioli RM
(2019) J Biomol Tech 30: 7-11
MeSH Terms: Animals, Brain, Brain Chemistry, Kidney, Mice, Molecular Imaging, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Show Abstract · Added March 3, 2020
Imaging mass spectrometry (IMS) has emerged as an important imaging modality because of its broad non-specific nature for molecular detection from highly complex samples. Within this broad field, new sub-categories of technologies have been developed incorporating many different molecular ionization processes. This article will focus on one of the major ionization processes, matrix-assisted laser desorption ionization (MALDI). IMS provides a critically important technology that brings new insight into complex biological samples and opens the door to new discoveries. Applications range widely, from fundamental studies in biology to specific clinical issues, all addressing the need to understand molecular spatial distributions at the tissue and cellular levels.
0 Communities
1 Members
0 Resources
7 MeSH Terms
Optimization of selective inversion recovery magnetization transfer imaging for macromolecular content mapping in the human brain.
Dortch RD, Bagnato F, Gochberg DF, Gore JC, Smith SA
(2018) Magn Reson Med 80: 1824-1835
MeSH Terms: Adult, Algorithms, Brain, Brain Chemistry, Brain Mapping, Female, Humans, Image Processing, Computer-Assisted, Magnetic Resonance Imaging, Male, Myelin Sheath, Phantoms, Imaging
Show Abstract · Added October 24, 2018
PURPOSE - To optimize a selective inversion recovery (SIR) sequence for macromolecular content mapping in the human brain at 3.0T.
THEORY AND METHODS - SIR is a quantitative method for measuring magnetization transfer (qMT) that uses a low-power, on-resonance inversion pulse. This results in a biexponential recovery of free water signal that can be sampled at various inversion/predelay times (t t ) to estimate a subset of qMT parameters, including the macromolecular-to-free pool-size-ratio (PSR), the R of free water (R ), and the rate of MT exchange (k ). The adoption of SIR has been limited by long acquisition times (≈4 min/slice). Here, we use Cramér-Rao lower bound theory and data reduction strategies to select optimal t /t combinations to reduce imaging times. The schemes were experimentally validated in phantoms, and tested in healthy volunteers (N = 4) and a multiple sclerosis patient.
RESULTS - Two optimal sampling schemes were determined: (i) a 5-point scheme (k estimated) and (ii) a 4-point scheme (k assumed). In phantoms, the 5/4-point schemes yielded parameter estimates with similar SNRs as our previous 16-point scheme, but with 4.1/6.1-fold shorter scan times. Pair-wise comparisons between schemes did not detect significant differences for any scheme/parameter. In humans, parameter values were consistent with published values, and similar levels of precision were obtained from all schemes. Furthermore, fixing k reduced the sensitivity of PSR to partial-volume averaging, yielding more consistent estimates throughout the brain.
CONCLUSIONS - qMT parameters can be robustly estimated in ≤1 min/slice (without independent measures of ΔB , B1+, and T ) when optimized t -t combinations are selected.
© 2018 International Society for Magnetic Resonance in Medicine.
0 Communities
3 Members
0 Resources
12 MeSH Terms
Inferring brain tissue composition and microstructure via MR relaxometry.
Does MD
(2018) Neuroimage 182: 136-148
MeSH Terms: Brain, Brain Chemistry, Humans, Magnetic Resonance Imaging, Models, Theoretical, Nerve Fibers, Myelinated, Neuroimaging
Show Abstract · Added March 5, 2020
MRI relaxometry is sensitive to a variety of tissue characteristics in a complex manner, which makes it both attractive and challenging for characterizing tissue. This article reviews the most common water proton relaxometry measures, T, T, and T, and reports on their development and current potential to probe the composition and microstructure of brain tissue. The development of these relaxometry measures is challenged by the need for suitably accurate tissue models, as well as robust acquisition and analysis methodologies. MRI relaxometry has been established as a tool for characterizing neural tissue, particular with respect to myelination, and the potential for further development exists.
Copyright © 2018 Elsevier Inc. All rights reserved.
0 Communities
1 Members
0 Resources
MeSH Terms
Decreased Rhes mRNA levels in the brain of patients with Parkinson's disease and MPTP-treated macaques.
Napolitano F, Booth Warren E, Migliarini S, Punzo D, Errico F, Li Q, Thiolat ML, Vescovi AL, Calabresi P, Bezard E, Morelli M, Konradi C, Pasqualetti M, Usiello A
(2017) PLoS One 12: e0181677
MeSH Terms: 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine, Aged, Aged, 80 and over, Animals, Bipolar Disorder, Brain Chemistry, Case-Control Studies, Female, GTP-Binding Proteins, Humans, Macaca mulatta, Male, Middle Aged, Parkinson Disease, Putamen, RNA, Messenger, Schizophrenia
Show Abstract · Added March 14, 2018
In rodent and human brains, the small GTP-binding protein Rhes is highly expressed in virtually all dopaminoceptive striatal GABAergic medium spiny neurons, as well as in large aspiny cholinergic interneurons, where it is thought to modulate dopamine-dependent signaling. Consistent with this knowledge, and considering that dopaminergic neurotransmission is altered in neurological and psychiatric disorders, here we sought to investigate whether Rhes mRNA expression is altered in brain regions of patients with Parkinson's disease (PD), Schizophrenia (SCZ), and Bipolar Disorder (BD), when compared to healthy controls (about 200 post-mortem samples). Moreover, we performed the same analysis in the putamen of non-human primate Macaca Mulatta, lesioned with the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Overall, our data indicated comparable Rhes mRNA levels in the brain of patients with SCZ and BD, and their respective healthy controls. In sharp contrast, the putamen of patients suffering from PD showed a significant 35% reduction of this transcript, compared to healthy subjects. Interestingly, in line with observations obtained in humans, we found 27% decrease in Rhes mRNA levels in the putamen of MPTP-treated primates. Based on the established inhibitory influence of Rhes on dopamine-related responses, we hypothesize that its striatal downregulation in PD patients and animal models of PD might represent an adaptive event of the dopaminergic system to functionally counteract the reduced nigrostriatal innervation.
0 Communities
1 Members
0 Resources
17 MeSH Terms
Assignment of the molecular origins of CEST signals at 2 ppm in rat brain.
Zhang XY, Xie J, Wang F, Lin EC, Xu J, Gochberg DF, Gore JC, Zu Z
(2017) Magn Reson Med 78: 881-887
MeSH Terms: Animals, Brain, Brain Chemistry, Image Processing, Computer-Assisted, Magnetic Resonance Imaging, Molecular Imaging, Proteins, Rats
Show Abstract · Added July 3, 2017
PURPOSE - Chemical exchange saturation transfer effects at 2 ppm (CEST@2ppm) in brain have previously been interpreted as originating from creatine. However, protein guanidino amine protons may also contribute to CEST@2ppm. This study aims to investigate the molecular origins and specificity of CEST@2ppm in brain.
METHODS - Two experiments were performed: (i) samples containing egg white albumin and creatine were dialyzed using a semipermeable membrane to demonstrate that proteins and creatine can be separated by this method; and (ii) tissue homogenates of rat brain with and without dialysis to remove creatine were studied to measure the relative contributions of proteins and creatine to CEST@2ppm.
RESULTS - The experiments indicate that dialysis can successfully remove creatine from proteins. Measurements on tissue homogenates show that, with the removal of creatine via dialysis, CEST@2ppm decreases to approximately 34% of its value before dialysis, which indicates that proteins and creatine have comparable contribution to the CEST@2ppm in brain. However, considering the contribution from peptides and amino acids to CEST@2ppm, creatine may have much less contribution to CEST@2ppm.
CONCLUSIONS - The contribution of proteins, peptides, and amino acids to CEST@2ppm cannot be neglected. The CEST@2ppm measurements of creatine in rat brain should be interpreted with caution. Magn Reson Med 78:881-887, 2017. © 2017 International Society for Magnetic Resonance in Medicine.
© 2017 International Society for Magnetic Resonance in Medicine.
0 Communities
3 Members
0 Resources
8 MeSH Terms
Trypsin and MALDI matrix pre-coated targets simplify sample preparation for mapping proteomic distributions within biological tissues by imaging mass spectrometry.
Zubair F, Laibinis PE, Swisher WG, Yang J, Spraggins JM, Norris JL, Caprioli RM
(2016) J Mass Spectrom 51: 1168-1179
MeSH Terms: Animals, Brain Chemistry, Molecular Imaging, Peptide Fragments, Peptide Mapping, Proteomics, Rats, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Trypsin
Show Abstract · Added March 10, 2017
Prefabricated surfaces containing α-cyano-4-hydroxycinnamic acid and trypsin have been developed to facilitate enzymatic digestion of endogenous tissue proteins prior to matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS). Tissue sections are placed onto slides that were previously coated with α-cyano-4-hydroxycinnamic acid and trypsin. After incubation to promote enzymatic digestion, the tissue is analyzed by MALDI IMS to determine the spatial distribution of the tryptic fragments. The peptides detected in the MALDI IMS dataset were identified by Liquid chromatography-tandem mass spectrometry/mass spectrometry. Protein identification was further confirmed by correlating the localization of unique tryptic fragments originating from common parent proteins. Using this procedure, proteins with molecular weights as large as 300 kDa were identified and their distributions were imaged in sections of rat brain. In particular, large proteins such as myristoylated alanine-rich C-kinase substrate (29.8 kDa) and spectrin alpha chain, non-erythrocytic 1 (284 kDa) were detected that are not observed without trypsin. The pre-coated targets simplify workflow and increase sample throughput by decreasing the sample preparation time. Further, the approach allows imaging at higher spatial resolution compared with robotic spotters that apply one drop at a time. Copyright © 2016 John Wiley & Sons, Ltd.
Copyright © 2016 John Wiley & Sons, Ltd.
1 Communities
3 Members
0 Resources
9 MeSH Terms
Neural stem cells sustain natural killer cells that dictate recovery from brain inflammation.
Liu Q, Sanai N, Jin WN, La Cava A, Van Kaer L, Shi FD
(2016) Nat Neurosci 19: 243-52
MeSH Terms: Aged, Aged, 80 and over, Animals, Brain, Brain Chemistry, Cell Proliferation, Cerebral Ventricles, Cytokines, Encephalitis, Encephalomyelitis, Autoimmune, Experimental, Female, Humans, Immune Tolerance, Interleukin-15, Killer Cells, Natural, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Multiple Sclerosis, Neural Stem Cells, Recovery of Function
Show Abstract · Added March 28, 2016
Recovery from organ-specific autoimmune diseases largely relies on the mobilization of endogenous repair mechanisms and local factors that control them. Natural killer (NK) cells are swiftly mobilized to organs targeted by autoimmunity and typically undergo numerical contraction when inflammation wanes. We report the unexpected finding that NK cells are retained in the brain subventricular zone (SVZ) during the chronic phase of multiple sclerosis in humans and its animal model in mice. These NK cells were found preferentially in close proximity to SVZ neural stem cells (NSCs) that produce interleukin-15 and sustain functionally competent NK cells. Moreover, NK cells limited the reparative capacity of NSCs following brain inflammation. These findings reveal that reciprocal interactions between NSCs and NK cells regulate neurorepair.
0 Communities
1 Members
0 Resources
22 MeSH Terms