scholarly journals Dissecting transcriptomic signatures of neuronal differentiation and maturation using iPSCs

2018 ◽  
Author(s):  
EE Burke ◽  
JG Chenoweth ◽  
JH Shin ◽  
L Collado-Torres ◽  
SK Kim ◽  
...  
Keyword(s):  
Neuronal Differentiation ◽  
Gene Networks ◽  
Splice Variants ◽  
Neuronal Cultures ◽  
Human Brain Tissue ◽  
Specific Expression ◽  
Individual Transcript ◽  
Human Neurons ◽  
Cell Type Specific Expression ◽  
Induced Pluripotent

SummaryHuman induced pluripotent stem cells (hiPSCs) are a powerful model of neural differentiation and maturation. We present a hiPSC transcriptomics resource on corticogenesis from 5 iPSC donor and 13 subclonal lines across nine time points over 5 broad conditions: self-renewal, early neuronal differentiation, neural precursor cells (NPCs), assembled rosettes, and differentiated neuronal cells that were validated using electrophysiology. We identified widespread changes in the expression of individual transcript features and their splice variants, gene networks, and global patterns of transcription. We next demonstrated that co-culturing human NPCs with rodent astrocytes resulted in mutually synergistic maturation, and that cell type-specific expression data can be extracted using only sequencing read alignments without potentially disruptive cell sorting. We lastly developed and validated a computational tool to estimate the relative neuronal maturity of iPSC-derived neuronal cultures and human brain tissue, which were maturationally heterogeneous but contained subsets of cells most akin to adult human neurons.

scholarly journals Influence of N-Arachidonoyl Dopamine and N-Docosahexaenoyl Dopamine on the Expression of Neurotrophic Factors in Neuronal Differentiated Cultures of Human Induced Pluripotent Stem Cells under Conditions of Oxidative Stress

Antioxidants ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 142
Author(s):  
Ekaterina Novosadova ◽  
Oleg Dolotov ◽  
Ludmila Inozemtseva ◽  
Ludmila Novosadova ◽  
Stanislav Antonov ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Neurotrophic Factors ◽  
Culture Medium ◽  
Neuronal Cultures ◽  
Neural Cells ◽  
Mrna Levels ◽  
Neuroprotective Effects ◽  
Human Neurons ◽  
Induced Pluripotent

Oxidative stress (OS) is implicated in the pathogenesis of several neurodegenerative diseases. We have previously shown that N-acyl dopamines (N-ADA and N-DDA) protect the neural cells of healthy donors and patients with Parkinson’s disease from OS. In this study, we assessed the effects of N-acyl dopamines on the expression of neurotrophic factors in human-induced pluripotent stem cell-derived neuronal cultures enriched with dopaminergic neurons under conditions of OS induced by hydrogen peroxide. We showed that hydrogen peroxide treatment increased BDNF but not GDNF mRNA levels, while it did not affect the secretion of corresponding proteins into the culture medium of these cells. Application of N-acyl dopamines promoted BDNF release into the culture medium. Under conditions of OS, N-DDA also increased TRKB, TRKC and RET mRNA levels. Furthermore, N-acyl dopamines prevented cell death 24 h after OS induction and promoted the expression of antioxidant enzymes GPX1, GPX7, SOD1, SOD2 and CAT, as well as reduced the BAX/BCL2 mRNA ratio. These findings indicate that stimulation of the expression of neurotrophic factors and their receptors may underlie the neuroprotective effects of N-acyl dopamines in human neurons.

Microarray-based discovery of highly expressed olfactory mucosal genes: potential roles in the various functions of the olfactory system

2003 ◽  
Vol 16 (1) ◽  
pp. 67-81 ◽  
Author(s):  
Mary Beth Genter ◽  
Paul P. Van Veldhoven ◽  
Anil G. Jegga ◽  
Bhuvana Sakthivel ◽  
Sue Kong ◽  
...  
Keyword(s):  
Neuronal Differentiation ◽  
Expression Pattern ◽  
Expression Patterns ◽  
Paraoxonase 1 ◽  
Specific Gene ◽  
Specific Expression ◽  
Mouse Cdna ◽  
Mouse Tissues ◽  
Cell Type Specific Expression ◽  
Gene Transcripts

We sought to gain a global view of tissue-specific gene expression in the olfactory mucosa (OM), the major site of neurogenesis and neuroregeneration in adult vertebrates, by examination of its overexpressed genes relative to that in 81 other developing and adult mouse tissues. We used a combination of statistical and fold-difference criteria to identify the top 269 cloned cDNAs from an array of 8,734 mouse cDNA elements on the Incyte Mouse GEM1 array. These clones, representing known and poorly characterized gene transcripts, were grouped according to their relative expression patterns across the other tissues and then further examined with respect to gene ontology categories. Approximately one-third of the 269 genes were also highly expressed in developing and/or adult central nervous system tissues. Several of these have been suggested or demonstrated to play roles in neurogenesis, neuronal differentiation, and/or neuronal migration, further suggesting that many of the unknown genes that share this expression pattern may play similar roles. Highly OM-specific genes included a palate, lung, and nasal epithelium carcinoma-associated gene ( Plunc); sphingosine phosphate lyase ( Sgpl1), and paraoxonase 1 ( Pon1). Cell-type-specific expression within OM was established using in situ hybridization for several representative expression pattern clusters. Using the ENSEMBL-assembled mouse genome and comparative genomics analyses to the human genome, we assigned many of the unknown expressed sequence tags (ESTs) and poorly characterized genes to either novel or known gene products and provided predictive classification. Further exploration of this database will provide additional insights into genes and pathways critical for olfactory neurogenesis, neuronal differentiation, olfaction, and mucosal defense.

scholarly journals Tissue and cell-type specific expression of a splice variant in the II-III cytoplasmic loop ofCacna1b

2019 ◽  
Author(s):  
Bunda Alexandra ◽  
LaCarubba Brianna ◽  
Akiki Marie ◽  
Andrade Arturo
Keyword(s):  
Motor Neurons ◽  
Splice Variants ◽  
Gene Encoding ◽  
Specific Expression ◽  
Cell Type Specific Expression ◽  
Alternatively Spliced ◽  
Cell Type Specific ◽  
Alternatively Spliced Exons ◽  
Cerebellar Cells

ABSTRACTPresynaptic CaV2.2 (N-type) channels are fundamental for transmitter release across the nervous system. The gene encoding CaV2.2 channels,Cacna1b, contains alternatively spliced exons that originate functionally distinct splice variants (e18a, e24a, e31a and 37a/37b). Alternative splicing of the cassette exon 18a generates two mRNA transcripts (+e18a-Cacna1band Δe18a-Cacna1b). In this study, using novel mouse genetic models and in situ hybridization (BaseScope™), we confirmed that +e18a-Cacna1bsplice variants are expressed in monoaminergic regions of midbrain. We expanded these studies and identified +e18a-Cacna1bmRNA in deep cerebellar cells and spinal cord motor neurons. Furthermore, we determined that +e18a-Cacna1bis enriched in cholecystokinin expressing interneurons. Our results provide key information to understand cell-specific functions of CaV2.2 channels.

scholarly journals Characterization of Promoter Function and Cell-Type-Specific Expression from Viral Vectors in the Nervous System

2000 ◽  
Vol 74 (23) ◽  
pp. 11254-11261 ◽  
Author(s):  
R. L. Smith ◽  
D. L. Traul ◽  
J. Schaack ◽  
G. H. Clayton ◽  
K. J. Staley ◽  
...  
Keyword(s):  
Dentate Gyrus ◽  
Viral Vectors ◽  
Neuronal Cultures ◽  
Specific Cell ◽  
Cell Type ◽  
Cmv Promoter ◽  
Specific Expression ◽  
Cell Type Specific Expression ◽  
Cell Type Specific ◽  
Selection Of

ABSTRACT Viral vectors have become important tools to effectively transfer genes into terminally differentiated cells, including neurons. However, the rational for selection of the promoter for use in viral vectors remains poorly understood. Comparison of promoters has been complicated by the use of different viral backgrounds, transgenes, and target tissues. Adenoviral vectors were constructed in the same vector background to directly compare three viral promoters, the human cytomegalovirus (CMV) immediate-early promoter, the Rous sarcoma virus (RSV) long terminal repeat, and the adenoviral E1A promoter, driving expression of the Escherichia coli lacZ gene or the gene for the enhanced green fluorescent protein. The temporal patterns, levels of expression, and cytotoxicity from the vectors were analyzed. In sensory neuronal cultures, the CMV promoter produced the highest levels of expression, the RSV promoter produced lower levels, and the E1A promoter produced limited expression. There was no evidence of cytotoxicity produced by the viral vectors. In vivo analyses following stereotaxic injection of the vector into the rat hippocampus demonstrated differences in the cell-type-specific expression from the CMV promoter versus the RSV promoter. In acutely prepared hippocampal brain slices, marked differences in the cell type specificity of expression from the promoters were confirmed. The CMV promoter produced expression in hilar regions and pyramidal neurons, with minimal expression in the dentate gyrus. The RSV promoter produced expression in dentate gyrus neurons. These results demonstrate that the selection of the promoter is critical for the success of the viral vector to express a transgene in specific cell types.

scholarly journals Human Brain Tissue Research

2021 ◽  
pp. 164-164
Author(s):  
Sujitha Suresh
Keyword(s):  
Animal Models ◽  
Brain Tissue ◽  
Neuronal Cultures ◽  
Valuable Insight ◽  
Human Stem Cells ◽  
Human Brain Tissue ◽  
Human Neurons ◽  
Science News ◽  
Human Brains ◽  
The Brain

Scientists can study human health conditions is through the use of human specimens. This approach is not only scientifically superior to the use of animal specimens, it is also more humane. Because of our commitment t0 advancing science without harming animals. NAVS is excited to see continued progress being made toward smarter, human-relevant research. An article published in Science News last month highlights the value of human biological samples and what can be done with these important scientific resources. The article focused on a project based in Seattle which aims to link patients who are willing to donate brain tissue removed during surgery with scientists and neurosurgeons. The brain tissue provided through this initiative is then experimented on at an amazingly quick pace. Just minutes after surgery, the brain tissue is transported to the research institute. Within an hour after surgery, it is sliced, preserved or kept alive for same-day experiments. Researchers are interested in learning more about the behavior and shape of the cells, as well as learning more about which genes were active in the cells of the brain tissue. The experiments being performed are also helping to explain why some drugs work differently in human brains than they do in mouse brains. For example, some human neurons have h-channel proteins, which help cells respond to electrical signals. These proteins are rare in mice, however. These channels can be impacted by drugs, which may account for differences observed between the species. The article further highlighted another important finding, noting that, "imprecise animal models have stymied research on schizophrenia, autism and Alzheimer's disease," which is "why studying live, human tissue is so critical." NAVS has long recognized the limitations of using animal models to study neurodegenerative conditions and has been helping fund alternatives for this purpose, using a different approach, one which relies on creating 3D neuronal cultures derived from human stem cells. Use of human-relevant models is providing valuable insight into human biology and disease and is producing results more quickly, cheaply and humanely than animal models.

scholarly journals A cell type‐specific expression map of NCoR1 and SMRT transcriptional co‐repressors in the mouse brain

2020 ◽  
Vol 528 (13) ◽  
pp. 2218-2238 ◽  
Author(s):  
Attilio Iemolo ◽  
Patricia Montilla‐Perez ◽  
I‐Chi Lai ◽  
Yinuo Meng ◽  
Syreeta Nolan ◽  
...  
Keyword(s):  
Mouse Brain ◽  
Cell Type ◽  
Specific Expression ◽  
Cell Type Specific Expression ◽  
A Cell ◽  
Cell Type Specific

scholarly journals High-content image-based analysis and proteomic profiling identifies Tau phosphorylation inhibitors in a human iPSC-derived glutamatergic neuronal model of tauopathy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chialin Cheng ◽  
Surya A. Reis ◽  
Emily T. Adams ◽  
Daniel M. Fass ◽  
Steven P. Angus ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Neural Progenitor Cells ◽  
Kinase Inhibitors ◽  
Ex Vivo ◽  
Clinical Investigation ◽  
Neuronal Model ◽  
Tau Phosphorylation ◽  
Neuronal Cultures ◽  
Proteomic Profiling ◽  
Induced Pluripotent

AbstractMutations in MAPT (microtubule-associated protein tau) cause frontotemporal dementia (FTD). MAPT mutations are associated with abnormal tau phosphorylation levels and accumulation of misfolded tau protein that can propagate between neurons ultimately leading to cell death (tauopathy). Recently, a p.A152T tau variant was identified as a risk factor for FTD, Alzheimer's disease, and synucleinopathies. Here we used induced pluripotent stem cells (iPSC) from a patient carrying this p.A152T variant to create a robust, functional cellular assay system for probing pathophysiological tau accumulation and phosphorylation. Using stably transduced iPSC-derived neural progenitor cells engineered to enable inducible expression of the pro-neural transcription factor Neurogenin 2 (Ngn2), we generated disease-relevant, cortical-like glutamatergic neurons in a scalable, high-throughput screening compatible format. Utilizing automated confocal microscopy, and an advanced image-processing pipeline optimized for analysis of morphologically complex human neuronal cultures, we report quantitative, subcellular localization-specific effects of multiple kinase inhibitors on tau, including ones under clinical investigation not previously reported to affect tau phosphorylation. These results demonstrate the potential for using patient iPSC-derived ex vivo models of tauopathy as genetically accurate, disease-relevant systems to probe tau biochemistry and support the discovery of novel therapeutics for tauopathies.

scholarly journals RBIO-06. IMPLEMENTATION OF HUMAN INDUCED PLURIPOTENT STEM CELL DERIVED CEREBRAL ORGANOIDS TO MODEL NORMAL TISSUE RADIORESPONSE

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii193-ii193
Author(s):  
Lawrence Bronk ◽  
Sanjay Singh ◽  
Riya Thomas ◽  
Luke Parkitny ◽  
Mirjana Maletic-Savatic ◽  
...  
Keyword(s):  
Stem Cell ◽  
Human Brain ◽  
Brain Tissue ◽  
Human Tissue ◽  
Model Systems ◽  
Human Brain Tissue ◽  
Post Irradiation ◽  
Mature Neurons ◽  
Induced Pluripotent ◽  
Effects Of Radiation

Abstract Treatment-related sequelae following cranial irradiation have life changing impacts for patients and their caregivers. Characterization of the basic response of human brain tissue to irradiation has been difficult due to a lack of preclinical models. The direct study of human brain tissue in vitro is becoming possible due to advances in stem cell biology, neuroscience, and tissue engineering with the development of organoids as novel model systems which enable experimentation with human tissue models. We sought to establish a cerebral organoid (CO) model to study the radioresponse of normal human brain tissue. COs were grown using human induced pluripotent stem cells and a modified Lancaster protocol. Compositional analysis during development of the COs showed expected populations of neurons and glia. We confirmed a population of microglia-like cells within the model positive for the makers Iba1 and CD68. After 2-months of maturation, COs were irradiated to 0, 10, and 20 Gy using a Shepard Mark-II Cs-137 irradiator and returned to culture. Subsets of COs were prepared for immunostaining at 30- and 70-days post-irradiation. To examine the effect of irradiation on the neural stem cell (NSC) population, sections were stained for SOX2 and Ki-67 expression denoting NSCs and proliferation respectively. Slides were imaged and scored using the CellProfiler software package. The percentage of proliferating NSCs 30-days post-irradiation was found to be significantly reduced for irradiated COs (5.7% (P=0.007) and 3.4% (P=0.001) for 10 and 20 Gy respectively) compared to control (12.7%). The reduction in the proliferating NSC population subsequently translated to a reduced population of NeuN-labeled mature neurons 70 days post-irradiation. The loss of proliferating NSCs and subsequent reduction in mature neurons demonstrates the long-term effects of radiation. Our initial results indicate COs will be a valuable model to study the effects of radiation therapy on normal and diseased human tissue.

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