scholarly journals Validating indicators of CNS disorders in a swine model of neurological disease

2019 ◽  
Author(s):  
Vicki J. Swier ◽  
Katherine A. White ◽  
David K. Meyerholz ◽  
Aude Chefdeville ◽  
Rajesh Khanna ◽  
...  
Keyword(s):  
Cellular Localization ◽  
Genetically Modified ◽  
Neurofibromatosis Type ◽  
Brain Regions ◽  
Swine Model ◽  
Behavioral Abnormalities ◽  
Human Disorders ◽  
Swine Tissue ◽  
Oligodendrocyte Development ◽  
Models Of Disease

AbstractGenetically modified porcine disease models are becoming increasingly important for studying molecular, physiological and pathological characteristics of human disorders. Given their limited history, there remains a great need for proven reagents in swine tissue. To provide a resource for neurological models of disease, we validated antibodies by immunohistochemistry for use in examining central nervous system (CNS) markers. To validate these tools in a relevant model, we utilized a recently developed miniswine model of neurofibromatosis type 1 (NF1). NF1 is a tumor predisposition disorder, presenting with different type of tumors. Additionally, neurological associated symptomologies may include chronic pain, cognitive impairment, and behavioral abnormalities, making this miniswine model an ideal candidate for validating CNS-relevant antibodies. We validate antibodies implicated in glial inflammation (CD68), oligodendrocyte development (NG2, O4, Olig2, and myelin PLP), and neuron differentiation and neurotransmission (doublecortin, GAD67, and tyrosine hydroxylase) by examining cellular localization and brain region specificity. Additionally, we confirm the utility of anti-GFAP, anti-Iba1, and anti-MBP antibodies, previously validated in swine, by testing their immunoreactivity across multiple brain regions in mutant NF1 samples. These validated immunostaining protocols for CNS markers provide a useful resource, furthering the utility of the genetically modified miniswine for translational and clinical applications.

Finding appropriate signal peptides for secretory production of recombinant glucarpidase: an in silico method

2021 ◽  
Vol 15 ◽  
Author(s):  
Omid Vakili ◽  
Seyyed Hossein Khatami ◽  
Amir Maleksabet ◽  
Ahmad Movahedpour ◽  
Saeed Ebrahimi Fana ◽  
...  
Keyword(s):  
Signal Peptide ◽  
In Silico ◽  
Cellular Localization ◽  
Clinical Aspects ◽  
Bioinformatics Analyses ◽  
E Coli ◽  
Accuracy And Precision ◽  
Extracellular Compartment ◽  
Human Disorders ◽  
High Doses

Aims: Bioinformatics analysis of suitable signal peptide for recombinant glucarpidase. Background: Methotrexate (MTX) is a general chemotherapeutic agent utilized to treat a variety of malignancies., woefully, its high doses can cause nephrotoxicity and subsequent defect in the process of MTX excretion. The recombinant form of glucarpidase, is produced by engineered E. coli and is a confirmed choice to overcoming this problem. Objective: In the present study, in silico analyses were performed to select suitable SPs for the secretion of recombinant glucarpidase in E. coli. Methods: The signal peptide website and UniProt database were employed to collect the SPs and protein sequences. In the next step, SignalP-5.0 helped us to predict the SPs and the position of cleavage sites. Moreover, physicochemical properties and solubility were evaluated using ProtParam and Protein-sol online software, and finally, ProtCompB was used to predict the final sub-cellular localization. Results: Luckily, all SPs could form soluble fusion proteins. At last, it was found that PPB and TIBA could translocate the glucarpidase into the extracellular compartment. Conclusion: This study showed that there are only 2 applicable SPs for the extracellular translocation of glucarpidase. Although the findings were remarkable with high degrees of accuracy and precision based on the utilization of bioinformatics analyses, additional experimental assessments are required to confirm and validate it. Recent patents revealed several inventions related to the clinical aspects of vaccine peptide against human disorders.

scholarly journals AUTOMATED RECOGNITION OF DEPRESSION FROM FEWER-SHOT LEANING IN RESTING-STATE fMRI WITH ReHo USING DEEP CONVOLUTIONAL NEURAL NETWORK

2021 ◽  
Author(s):  
MI LI ◽  
JINYU ZHANG ◽  
QIAN ZHAI ◽  
JIAMING KANG ◽  
SHENGFU LU ◽  
...  
Keyword(s):  
Resting State ◽  
Resting State Fmri ◽  
Brain Regions ◽  
Automated Recognition ◽  
Convolutional Networks ◽  
Behavioral Abnormalities ◽  
Kernel Extreme Learning Machine ◽  
Learning Machine ◽  
Functional Areas

Up to now, there is still the absence of research about depression recognition using resting-state functional magnetic resonance imaging (rest_fMRI) and deep learning. Previous studies have shown that regional homogeneity (ReHo) of rest_fMRI (rest_ReHo_fMRI) is a characterization of the functional synchronization of adjacent voxels in brain regions, and the mental and behavioral abnormalities in depression are due to an imbalance of ReHo synchronization in some brain functional areas. Accordingly, this paper presents a method for depression recognition using rest_ReHo_fMRI. First, the rest_ReHo_fMRI is extracted from the preprocessed rest-fMRI by calculation. Then, deep convolutional networks (such as VGG16) pretrained on ImageNet are used to automatically complete extracting the classification features from rest_ReHo_fMRI. Finally, the Kernel Extreme Learning Machine (KELM) was used to classify the depression. The results of the test set show that the proposed method achieves 89.07% in sensitivity and 89.74% in specificity. This study suggests that features of rest_ReHo_fMRI can be used as biomarkers to distinguish depression from normal people.

scholarly journals Novel Insights into the Cellular Localization and Regulation of the Autophagosomal Proteins LC3A, LC3B and LC3C

Cells ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 2315
Author(s):  
Marius W. Baeken ◽  
Katja Weckmann ◽  
Philip Diefenthäler ◽  
Jan Schulte ◽  
Kamran Yusifli ◽  
...  
Keyword(s):  
Cellular Localization ◽  
Human Fibroblasts ◽  
Sirtuin 1 ◽  
Autophagic Flux ◽  
Molecular Function ◽  
Knock Down ◽  
Human Disorders ◽  
Total Protein Expression ◽  
Protein Lipidation ◽  
Starvation Conditions

Macroautophagy is a conserved degradative process for maintaining cellular homeostasis and plays a key role in aging and various human disorders. The microtubule-associated protein 1A/1B light chain 3B (MAP1LC3B or LC3B) is commonly analyzed as a key marker for autophagosomes and as a proxy for autophagic flux. Three paralogues of the LC3 gene exist in humans: LC3A, LC3B and LC3C. The molecular function, regulation and cellular localization of LC3A and LC3C have not been investigated frequently, even if a similar function to that described for LC3B appears likely. Here, we have selectively decapacitated LC3B by three separate strategies in primary human fibroblasts and analyzed the evoked effects on LC3A, LC3B and LC3C in terms of their cellular distribution and co-localization with p62, a ubiquitin and autophagy receptor. First, treatment with pharmacological sirtuin 1 (SIRT1) inhibitors to prevent the translocation of LC3B from the nucleus into the cytosol induced an increase in cytosolic LC3C, a heightened co-localization of LC3C with p62, and an increase LC3C-dependent autophagic flux as assessed by protein lipidation. Cytosolic LC3A, however, was moderately reduced, but also more co-localized with p62. Second, siRNA-based knock-down of SIRT1 broadly reproduced these findings and increased the co-localization of LC3A and particularly LC3C with p62 in presumed autophagosomes. These effects resembled the effects of pharmacological sirtuin inhibition under normal and starvation conditions. Third, siRNA-based knock-down of total LC3B in cytosol and nucleus also induced a redistribution of LC3C as if to replace LC3B in the nucleus, but only moderately affected LC3A. Total protein expression of LC3A, LC3B, LC3C, GABARAP and GABARAP-L1 following LC3B decapacitation was unaltered. Our data indicate that nuclear trapping and other causes of LC3B functional loss in the cytosol are buffered by LC3A and actively compensated by LC3C, but not by GABARAPs. The biological relevance of the potential functional compensation of LC3B decapacitation by LC3C and LC3A warrants further study.

scholarly journals Rapid Acting Antidepressants in Chronic Stress Models: Molecular and Cellular Mechanisms

2017 ◽  
Vol 1 ◽  
pp. 247054701769731 ◽  
Author(s):  
Brendan D. Hare ◽  
Sriparna Ghosal ◽  
Ronald S. Duman
Keyword(s):  
Selective Serotonin Reuptake Inhibitors ◽  
Morphological Changes ◽  
Time Lag ◽  
Treatment Strategies ◽  
The United States ◽  
Brain Regions ◽  
Cellular Mechanisms ◽  
Behavioral Abnormalities ◽  
Suitable Treatment ◽  
Stress Models

Stress-associated disorders, including depression and anxiety, impact nearly 20% of individuals in the United States. The social, health, and economic burden imposed by stress-associated disorders requires in depth research efforts to identify suitable treatment strategies. Traditional medications (e.g., selective serotonin reuptake inhibitors, monoamine oxidase inhibitors) have significant limitations, notably a time lag for therapeutic response that is compounded by low rates of efficacy. Excitement over ketamine, a rapid acting antidepressant effective in treatment resistant patients, is tempered by transient dissociative and psychotomimetic effects, as well as abuse potential. Rodent stress models are commonly used to produce behavioral abnormalities that resemble those observed in stress-associated disorders. Stress models also produce molecular and cellular morphological changes in stress sensitive brain regions, including the prefrontal cortex and hippocampus that resemble alterations observed in depression. Rapid acting antidepressants such as ketamine can rescue stress-associated morphological and behavioral changes in rodent models. Here, we review the literature supporting a role for rapid acting antidepressants in opposing the effects of stress, and summarize research efforts seeking to elucidate the molecular, cellular, and circuit level targets of these agents.

scholarly journals To Ubiquitinate or Not to Ubiquitinate: TRIM17 in Cell Life and Death

Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1235
Author(s):  
Meenakshi Basu-Shrivastava ◽  
Alina Kozoriz ◽  
Solange Desagher ◽  
Iréna Lassot
Keyword(s):  
Ubiquitin Ligase ◽  
E3 Ubiquitin Ligase ◽  
Brain Regions ◽  
Cell Physiology ◽  
Life And Death ◽  
Cellular Processes ◽  
Cell Life ◽  
Ubiquitin Ligase Activity ◽  
Human Disorders ◽  
Trim Proteins

TRIM17 is a member of the TRIM family, a large class of RING-containing E3 ubiquitin-ligases. It is expressed at low levels in adult tissues, except in testis and in some brain regions. However, it can be highly induced in stress conditions which makes it a putative stress sensor required for the triggering of key cellular responses. As most TRIM members, TRIM17 can act as an E3 ubiquitin-ligase and promote the degradation by the proteasome of substrates such as the antiapoptotic protein MCL1. Intriguingly, TRIM17 can also prevent the ubiquitination of other proteins and stabilize them, by binding to other TRIM proteins and inhibiting their E3 ubiquitin-ligase activity. This duality of action confers several pivotal roles to TRIM17 in crucial cellular processes such as apoptosis, autophagy or cell division, but also in pathological conditions as diverse as Parkinson’s disease or cancer. Here, in addition to recent data that endorse this duality, we review what is currently known from public databases and the literature about TRIM17 gene regulation and expression, TRIM17 protein structure and interactions, as well as its involvement in cell physiology and human disorders.

scholarly journals Substantial acetylcholine reduction in multiple brain regions of Mecp2-deficient female rats and associated behavioral abnormalities

PLoS ONE ◽  
2021 ◽  
Vol 16 (10) ◽  
pp. e0258830
Author(s):  
Hiroyasu Murasawa ◽  
Hiroyuki Kobayashi ◽  
Jun Imai ◽  
Takahiko Nagase ◽  
Hitomi Soumiya ◽  
...  
Keyword(s):  
Plasma Insulin ◽  
Gene Deletion ◽  
Neurodevelopmental Disorder ◽  
Brain Regions ◽  
Female Rats ◽  
Future Research ◽  
Dominant Inheritance ◽  
Lower Plasma ◽  
Robust Model ◽  
Behavioral Abnormalities

Rett syndrome (RTT) is a neurodevelopmental disorder with X-linked dominant inheritance caused mainly by mutations in the methyl-CpG-binding protein 2 (MECP2) gene. The effects of various Mecp2 mutations have been extensively assessed in mouse models, but none adequately mimic the symptoms and pathological changes of RTT. In this study, we assessed the effects of Mecp2 gene deletion on female rats (Mecp2+/−) and found severe impairments in social behavior [at 8 weeks (w), 12 w, and 23 w of age], motor function [at 16 w and 26 w], and spatial cognition [at 29 w] as well as lower plasma insulin-like growth factor (but not brain-derived neurotrophic factor) and markedly reduced acetylcholine (30%–50%) in multiple brain regions compared to female Mecp2+/+ rats [at 29 w]. Alternatively, changes in brain monoamine levels were relatively small, in contrast to reports on mouse Mecp2 mutants. Female Mecp2-deficient rats express phenotypes resembling RTT and so may provide a robust model for future research on RTT pathobiology and treatment.

scholarly journals Cellular basis of memory for addiction

2013 ◽  
Vol 15 (4) ◽  
pp. 431-443 ◽  
Keyword(s):  
Drug Addiction ◽  
Biological Process ◽  
Brain Regions ◽  
Loss Of Control ◽  
Drug Induced ◽  
Cellular Basis ◽  
Behavioral Abnormalities ◽  
Adaptive Mechanisms ◽  
Neurotrophic Factor Signaling ◽  
Behavioral Memory

DESPITE THE IMPORTANCE OF NUMEROUS PSYCHOSOCIAL FACTORS, AT ITS CORE, DRUG ADDICTION INVOLVES A BIOLOGICAL PROCESS: the ability of repeated exposure to a drug of abuse to induce changes in a vulnerable brain that drive the compulsive seeking and taking of drugs, and loss of control over drug use, that define a state of addiction. Here, we review the types of molecular and cellular adaptations that occur in specific brain regions to mediate addiction-associated behavioral abnormalities. These include alterations in gene expression achieved in part via epigenetic mechanisms, plasticity in the neurophysiological functioning of neurons and synapses, and associated plasticity in neuronal and synaptic morphology mediated in part by altered neurotrophic factor signaling. Each of these types of drug-induced modifications can be viewed as a form of "cellular or molecular memory." Moreover, it is striking that most addiction-related forms of plasticity are very similar to the types of plasticity that have been associated with more classic forms of "behavioral memory," perhaps reflecting the finite repertoire of adaptive mechanisms available to neurons when faced with environmental challenges. Finally, addiction-related molecular and cellular adaptations involve most of the same brain regions that mediate more classic forms of memory, consistent with the view that abnormal memories are important drivers of addiction syndromes. The goal of these studies which aim to explicate the molecular and cellular basis of drug addiction is to eventually develop biologically based diagnostic tests, as well as more effective treatments for addiction disorders.

scholarly journals Lineage recording reveals dynamics of cerebral organoid regionalization

2020 ◽  
Author(s):  
Zhisong He ◽  
Tobias Gerber ◽  
Ashley Maynard ◽  
Akanksha Jain ◽  
Rebecca Petri ◽  
...  
Keyword(s):  
Time Course ◽  
Light Sheet ◽  
Brain Regions ◽  
Patient Specific ◽  
Light Sheet Microscopy ◽  
Brain Organoid ◽  
Induced Pluripotent ◽  
Models Of Disease ◽  
Fate Restriction ◽  
Lineage Trees

Diverse regions develop within cerebral organoids generated from human induced pluripotent stem cells (iPSCs), however it has been a challenge to understand the lineage dynamics associated with brain regionalization. Here we establish an inducible lineage recording system that couples reporter barcodes, inducible CRISPR/Cas9 scarring, and single-cell transcriptomics to analyze lineage relationships during cerebral organoid development. We infer fate-mapped whole organoid phylogenies over a scarring time course, and reconstruct progenitor-neuron lineage trees within microdissected cerebral organoid regions. We observe increased fate restriction over time, and find that iPSC clones used to initiate organoids tend to accumulate in distinct brain regions. We use lineage-coupled spatial transcriptomics to resolve lineage locations as well as confirm clonal enrichment in distinctly patterned brain regions. Using long term 4-D light sheet microscopy to temporally track nuclei in developing cerebral organoids, we link brain region clone enrichment to positions in the neuroectoderm, followed by local proliferation with limited migration during neuroepithelial formation. Our data sheds light on how lineages are established during brain organoid regionalization, and our techniques can be adapted in any iPSC-derived cell culture system to dissect lineage alterations during perturbation or in patient-specific models of disease.

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