scholarly journals Function of Oncogene Mycn in Adult Neurogenesis and Oligodendrogenesis

2021 ◽  
Author(s):  
Jiao Chen ◽  
Zhonghui Guan
Keyword(s):  
Subventricular Zone ◽  
Brain Regions ◽  
Embryonic Brain ◽  
Subgranular Zone ◽  
Wild Type ◽  
Rt Pcr ◽  
Proliferating Cells ◽  
Adult Mice ◽  
Nerve System ◽  
Embryonic Brain Development

AbstractHuman MYCN is an oncogene amplified in neuroblastoma and many other tumors. Both human MYCN and mouse Mycn genes are important in embryonic brain development, but their functions in adult healthy nerve system are completely unknown. Here, with Mycn-eGFP mice and quantitative RT-PCR, we found that Mycn was expressed in specific brain regions of young adult mice, including subventricular zone (SVZ), subgranular zone (SGZ), olfactory bulb (OB), subcallosal zone (SCZ), and corpus callosum (CC). With immunohistochemistry (IHC), we found that many Mycn-expressing cells expressed neuroblast marker doublecortin (DCX) and proliferation marker Ki67. With Dcx-creER and Mki67-creER mouse lines, we fate mapped Dcx-expressing neuroblasts and Mki67-expressing proliferation cells, along with deleting Mycn from these cells in adult mice. We found that knocking out Mycn from adult neuroblasts or proliferating cells significantly reduced cells in proliferation in SVZ, SGZ, OB, SCZ, and CC. We also demonstrated that the Mycn-deficient neuroblasts in SGZ matured quicker than wild-type neuroblasts, and that Mycn-deficient proliferating cells were more likely to survive in SVZ, SGZ, OB, SCZ, and CC compared to wild type. Thus, our results demonstrate that, in addition to causing tumors in the nervous system, oncogene Mycn has a crucial function in neurogenesis and oligodendrogenesis in adult healthy brain.

scholarly journals Ekspresi Level Gen mRNA Protein Ekstraseluler Otak Embrio Mencit Black-6 UK-12 Akibat Induksi 2-Methoxyethanol : Analisis secara Real Time RT-PCR

2010 ◽  
Vol 15 (2) ◽  
pp. 171-179
Author(s):  
Yulia Irnidayanti ◽  
Win Darmanto ◽  
Agus Abadi
Keyword(s):  
Brain Development ◽  
Intermediate Filaments ◽  
Cell Types ◽  
Specific Cell ◽  
Embryonic Brain ◽  
Rt Pcr ◽  
Autoimmune Inflammation ◽  
Central Nervous Systems ◽  
And Control ◽  
Embryonic Brain Development

The aim of this research was to investigate impact of 2-methoxyethanol, a major industrial chemical, and its individual metabolites on the expression DNA of the embryonic brain development of black-6 mice. The expression levels mRNA protein of GAPDH, Fibronectin, tenascin, vimentin, Neurofilamen, NCam between brain embrio treatment with 2-ME at gestation day 12 and Embryo control were achieved. The Electroforesis DNA on brain Embryonic day 12 showed that there were expression of GAPDH (447bp), Fibronectin (462bp), NCAM (293 bp), Tenascin (416bp), Vimentin (327), Neurofilamen high (301bp), Neurofilamen medium (289bp), Neurofilamen low (398bp). This Data not showed. The expression of level of mRNA for protein Vimentin at embryonic brain treatment at GD-12 is 487 copies, meanwhile on the embryonic brain control is 209 copies. This expression is tendency very higher than control. Another level of mRNA for protein fibronectin, NCAM, Tenascin, Neurofilament were tendency not difference between embryonic brain treatments and control. Intermediate filaments, vimentin, is found in specific cell types in the developing and adult central nervous systems (CNS), particularly astrocytes. Recently, found that vimentin immunoreactivities were increased in astrocytes and/or macrophages in the spinal cords of rats with autoimmune inflammation). So that The higher level mRNA for protein vimentin caused by effect 2-methoxyethanol. Vimentin contribute to the repair of brain through the migration of activated cells and increased level vimentin at embryionic brain treatment with 2-ME.

scholarly journals Altered Expression of GABAergic Markers in the Forebrain of Young and Adult Engrailed-2 Knockout Mice

Genes ◽  
2020 ◽  
Vol 11 (4) ◽  
pp. 384
Author(s):  
Giovanni Provenzano ◽  
Angela Gilardoni ◽  
Marika Maggia ◽  
Mattia Pernigo ◽  
Paola Sgadò ◽  
...  
Keyword(s):  
Gaba Receptors ◽  
Gaba Receptor ◽  
Brain Regions ◽  
Autism Spectrum ◽  
Gabaergic Interneuron ◽  
Gabaergic Interneurons ◽  
Rt Pcr ◽  
Altered Expression ◽  
Gabaergic Neurotransmission ◽  
Adult Mice

Impaired function of GABAergic interneurons, and the subsequent alteration of excitation/inhibition balance, is thought to contribute to autism spectrum disorders (ASD). Altered numbers of GABAergic interneurons and reduced expression of GABA receptors has been detected in the brain of ASD subjects and mouse models of ASD. We previously showed a reduced expression of GABAergic interneuron markers parvalbumin (PV) and somatostatin (SST) in the forebrain of adult mice lacking the Engrailed2 gene (En2-/- mice). Here, we extended this analysis to postnatal day (P) 30 by using in situ hybridization, immunohistochemistry, and quantitative RT-PCR to study the expression of GABAergic interneuron markers in the hippocampus and somatosensory cortex of En2-/- and wild type (WT) mice. In addition, GABA receptor subunit mRNA expression was investigated by quantitative RT-PCR in the same brain regions of P30 and adult En2-/- and WT mice. As observed in adult animals, PV and SST expression was decreased in En2-/- forebrain of P30 mice. The expression of GABA receptor subunits (including the ASD-relevant Gabrb3) was also altered in young and adult En2-/- forebrain. Our results suggest that GABAergic neurotransmission deficits are already evident at P30, confirming that neurodevelopmental defects of GABAergic interneurons occur in the En2 mouse model of ASD.

scholarly journals Temporally and Spatially Regulated Expression of the Linker Histone H1fx During Mouse Development

2017 ◽  
Vol 65 (9) ◽  
pp. 513-530 ◽  
Author(s):  
Keiko Ichihara-Tanaka ◽  
Kenji Kadomatsu ◽  
Satoshi Kishida
Keyword(s):  
Subventricular Zone ◽  
Immunohistochemical Analysis ◽  
Linker Histone ◽  
Developmental Changes ◽  
Subgranular Zone ◽  
Embryonic Cells ◽  
Mouse Development ◽  
Ki 67 ◽  
Adult Mice ◽  
Regulated Expression

The linker histone H1fx is the least characterized member of the H1 family. To investigate the developmental changes of H1fx, we performed an immunohistochemical analysis of its expression pattern from embryos to adult mice. We found that H1fx was highly expressed during gastrulation, and was positive in all embryonic germ layers between E8.5 and E10.5, which mostly overlapped with the expression of the proliferation marker Ki-67. Neural and mesenchyme tissues strongly expressed H1fx at E10.5. H1fx expression began to be restricted at around E12.5. Western blot analysis of brain tissues demonstrated that the total expression level of H1fx gradually decreased with time from E12.5 to adulthood, whereas H1f0 was increased over this period. In adult mice, H1fx was restrictively expressed at the hypothalamus, subventricular zone, subgranular zone, medulla of the adrenal grand, islets of Langerhans, and myenteric plexus. Taken together, these data suggest that H1fx is preferentially expressed in immature embryonic cells and plays some roles in cells with neural properties.

scholarly journals Wnt/β-catenin signalling promotes more effective fracture healing in aged mice than in adult mice by inducing angiogenesis and cell differentiation

2021 ◽  
Vol 104 (2) ◽  
pp. 003685042110132
Author(s):  
Daocheng Liu ◽  
Sihao He ◽  
Sixu Chen ◽  
Lei Yang ◽  
Jiazhi Yang ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Fracture Healing ◽  
Wild Type ◽  
Rt Pcr ◽  
Aged Patients ◽  
Aged Mice ◽  
Adult Mice ◽  
Diaphyseal Bone ◽  
Trap Staining ◽  
Polymerase Chain

To investigate whether activating the Wnt/β-catenin signalling pathway differentially promotes fracture healing in aged and adult individuals. CatnbTM2Kem, Catnblox(ex3) and wild-type adult and aged mice were used in this study. The femur was electroporated through a hole with a diameter of 0.6 mm. On the 7th, 14th and 21st days after fracture establishment, repair of the femoral diaphyseal bone was examined using X-ray and CT, the levels of mRNAs related to Wnt/β-catenin signalling were detected using real-time polymerase chain reaction (RT-PCR), and angiogenesis and cell differentiation were observed using immunohistochemistry. The numbers of osteoclasts were determined by TRAP staining. Wnt/β-catenin activation accelerated fracture healing in adult mice, with more pronounced effects on aged mice. Compared with wild-type mice at the corresponding ages, Wnt/β-catenin signalling activation induced higher levels of angiogenesis and cell differentiation in aged mice than in adult mice and promoted fracture healing. The administration of medications targeting Wnt/β-catenin signalling to aged patients may accelerate fracture healing to a greater extent.

scholarly journals The Cloning and Molecular Analysis of pawn-B in Paramecium tetraurelia

Genetics ◽  
2000 ◽  
Vol 155 (3) ◽  
pp. 1105-1117 ◽  
Author(s):  
W John Haynes ◽  
Kit-Yin Ling ◽  
Robin R Preston ◽  
Yoshiro Saimi ◽  
Ching Kung
Keyword(s):  
Genomic Library ◽  
Open Reading Frame ◽  
Paramecium Tetraurelia ◽  
Wild Type ◽  
Rt Pcr ◽  
Reading Frame ◽  
Close Proximity ◽  
In The Wild ◽  
Dna Fragment ◽  
Alternative Sites

Abstract Pawn mutants of Paramecium tetraurelia lack a depolarization-activated Ca2+ current and do not swim backward. Using the method of microinjection and sorting a genomic library, we have cloned a DNA fragment that complements pawn-B (pwB/pwB). The minimal complementing fragment is a 798-bp open reading frame (ORF) that restores the Ca2+ current and the backward swimming when expressed. This ORF contains a 29-bp intron and is transcribed and translated. The translated product has two putative transmembrane domains but no clear matches in current databases. Mutations in the available pwB alleles were found within this ORF. The d4-95 and d4-96 alleles are single base substitutions, while d4-662 (previously pawn-D) harbors a 44-bp insertion that matches an internal eliminated sequence (IES) found in the wild-type germline DNA except for a single C-to-T transition. Northern hybridizations and RT-PCR indicate that d4-662 transcripts are rapidly degraded or not produced. A second 155-bp IES in the wild-type germline ORF excises at two alternative sites spanning three asparagine codons. The pwB ORF appears to be separated from a 5′ neighboring ORF by only 36 bp. The close proximity of the two ORFs and the location of the pwB protein as indicated by GFP-fusion constructs are discussed.

scholarly journals FGFR3 overexpression is a useful detection tool for FGFR3 fusions and sequence variations in glioma

2020 ◽  
Author(s):  
Jens Schittenhelm ◽  
Lukas Ziegler ◽  
Jan Sperveslage ◽  
Michel Mittelbronn ◽  
David Capper ◽  
...  
Keyword(s):  
Coiled Coil ◽  
Growth Factor Receptor ◽  
Gene Mutations ◽  
Diagnostic Tools ◽  
Wild Type ◽  
Rt Pcr ◽  
Fgfr3 Gene ◽  
Detection Tool ◽  
Sequence Variations ◽  
Gene Panel Sequencing

Abstract Background Fibroblast growth factor receptor (FGFR) inhibitors are currently used in clinical development. A subset of glioblastomas carries gene fusion of FGFR3 and transforming acidic coiled-coil protein 3. The prevalence of other FGFR3 alterations in glioma is currently unclear. Methods We performed RT-PCR in 101 glioblastoma samples to detect FGFR3-TACC3 fusions (“RT-PCR cohort”) and correlated results with FGFR3 immunohistochemistry (IHC). Further, we applied FGFR3 IHC in 552 tissue microarray glioma samples (“TMA cohort”) and validated these results in two external cohorts with 319 patients. Gene panel sequencing was carried out in 88 samples (“NGS cohort”) to identify other possible FGFR3 alterations. Molecular modeling was performed on newly detected mutations. Results In the “RT-PCR cohort,” we identified FGFR3-TACC3 fusions in 2/101 glioblastomas. Positive IHC staining was observed in 73/1024 tumor samples of which 10 were strongly positive. In the “NGS cohort,” we identified FGFR3 fusions in 9/88 cases, FGFR3 amplification in 2/88 cases, and FGFR3 gene mutations in 7/88 cases in targeted sequencing. All FGFR3 fusions and amplifications and a novel FGFR3 K649R missense mutation were associated with FGFR3 overexpression (sensitivity and specificity of 93% and 95%, respectively, at cutoff IHC score > 7). Modeling of these data indicated that Tyr647, a residue phosphorylated as a part of FGFR3 activation, is affected by the K649R mutation. Conclusions FGFR3 IHC is a useful screening tool for the detection of FGFR3 alterations and could be included in the workflow for isocitrate dehydrogenase (IDH) wild-type glioma diagnostics. Samples with positive FGFR3 staining could then be selected for NGS-based diagnostic tools.

scholarly journals STABILITY OF DNA IN PURKINJE CELL NUCLEI OF THE MOUSE

1974 ◽  
Vol 63 (2) ◽  
pp. 665-674 ◽  
Author(s):  
V. Mareš ◽  
B. Schultze ◽  
W. Maurer
Keyword(s):  
Young Adult ◽  
Purkinje Cell ◽  
Nuclear Dna ◽  
Brain Regions ◽  
Nuclear Volume ◽  
High Dose ◽  
Prenatal Period ◽  
Cell Nuclei ◽  
Grain Number ◽  
Adult Mice

Neurons of the mouse were labeled with [3H]thymidine during their prenatal period of proliferation. The 3H activity of the Purkinje cell nuclei was then studied autoradiographically 8, 25, 55, and 90 days after birth. The measured grain number per nucleus decreased by about 14% between the 8th and 25th postnatal days and then remained constant up to 90 days. There was no significant decrease of the 3H activity of the Purkinje cell nuclei after correction of the measured grain number per nucleus for increasing nuclear volume of the growing Purkinje cells and for the influence of [3H]ß self-absorption in the material of the sections. Injection of a high dose of [3H]thymidine into young adult mice did not result in 3H labeling of either Purkinje or other neurons in other brain regions. The results agree with the concept of metabolic stability of nuclear DNA. "Metabolic" DNA could not be observed in these experiments.

scholarly journals MRG15 Regulates Embryonic Development and Cell Proliferation

2005 ◽  
Vol 25 (8) ◽  
pp. 2924-2937 ◽  
Author(s):  
Kaoru Tominaga ◽  
Bhakti Kirtane ◽  
James G. Jackson ◽  
Yuji Ikeno ◽  
Takayoshi Ikeda ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Embryonic Development ◽  
Chromatin Remodeling ◽  
Histone Deacetylases ◽  
Globin Gene ◽  
Immunohistochemical Analysis ◽  
Wild Type ◽  
Proliferating Cells ◽  
Globin Promoter

ABSTRACT MRG15 is a highly conserved protein, and orthologs exist in organisms from yeast to humans. MRG15 associates with at least two nucleoprotein complexes that include histone acetyltransferases and/or histone deacetylases, suggesting it is involved in chromatin remodeling. To study the role of MRG15 in vivo, we generated knockout mice and determined that the phenotype is embryonic lethal, with embryos and the few stillborn pups exhibiting developmental delay. Immunohistochemical analysis indicates that apoptosis in Mrg15 − / − embryos is not increased compared with wild-type littermates. However, the number of proliferating cells is significantly reduced in various tissues of the smaller null embryos compared with control littermates. Cell proliferation defects are also observed in Mrg15 − / − mouse embryonic fibroblasts. The hearts of the Mrg15 − / − embryos exhibit some features of hypertrophic cardiomyopathy. The increase in size of the cardiomyocytes is most likely a response to decreased growth of the cells. Mrg15 − / − embryos appeared pale, and microarray analysis revealed that α-globin gene expression was decreased in null versus wild-type embryos. We determined by chromatin immunoprecipitation that MRG15 was recruited to the α-globin promoter during dimethyl sulfoxide-induced mouse erythroleukemia cell differentiation. These findings demonstrate that MRG15 has an essential role in embryonic development via chromatin remodeling and transcriptional regulation.

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