Complex Effects of the ZSCAN21 Transcription Factor on Transcriptional Regulation of α-Synuclein in Primary Neuronal Cultures andin Vivo

Целью настоящей работы стало исследование влияния лактоферрина (Лф) человека на экспрессию транскрипционного фактора c-Fos в первичных нейрональных культурах после физиологической стимуляции, определение клеточной локализации Лф человека и возможной колокализации экзогенного белка с индуцированной экспрессией c-Fos. Методы. Первичные диссоциированные клеточные культуры получали из гиппокампа головного мозга новорожденных мышей (Р0-Р1) линии С57Вl/6. Индукцию экспрессии белка c-Fos в клетках осуществляли путем трехкратного добавления 50 мМ KСl в культуральную среду на 8-й день культивирования in vitro. Анализ содержания c-Fos проводили иммунофлюоресцентным методом через 2 часа после стимуляции. Результаты. Лф детектировался как в цитоплазме, так и в ядрах отдельных клеток культуры после стимуляции KСl. В ядрах некоторых клеток была выявлена колокализация включения Лф и экспрессии c-Fos. Было обнаружено, что предварительное введение Лф в культуральную среду увеличивало количество клеток, экспрессирующих c-Fos после добавления 50 мМ KСl. The aims of this research were 1) to study the effect of human lactoferrin (Lf) on the expression of the c-Fos transcription factor in primary neuronal cultures after physiological stimulation; 2) to determine the cellular localization of human Lf and possible colocalization of an exogenous protein with induced c-Fos expression. Methods. Primary dissociated cell cultures were obtained from the hippocampus of newborn C57Bl/6 mice (P0-P1). The expression of c-Fos was induced by addition of 50 mM KCl to the culture medium at 8 day in vitro. c-Fos content was analyzed by immunofluorescence 2 hrs after stimulation. Results. Lf was detected in cytoplasm and in nuclei after stimulation KCl. Lf inclusion and c-Fos expression were colocalized in the nuclei of some cells. Thus, results showed that pretreatment with Lf led to increase in the number of cells expressing c-Fos after exposure to 50 mM KCl.

Download Full-text

Faculty Opinions recommendation of Post-transcriptional regulation of the Sef1 transcription factor controls the virulence of Candida albicans in its mammalian host.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.717963075.793465696 ◽

2012 ◽

Author(s):

Malcolm Whiteway

Keyword(s):

Transcription Factor ◽

Transcriptional Regulation ◽

Candida Albicans ◽

Mammalian Host ◽

Post Transcriptional Regulation

Download Full-text

Validation of long-term primary neuronal cultures and network activity through the integration of reversibly bonded microbioreactors and MEA substrates

Biotechnology and Bioengineering ◽

10.1002/bit.23310 ◽

2011 ◽

Vol 109 (1) ◽

pp. 166-175 ◽

Cited By ~ 16

Author(s):

Emilia Biffi ◽

Andrea Menegon ◽

Francesco Piraino ◽

Alessandra Pedrocchi ◽

Gianfranco B. Fiore ◽

...

Keyword(s):

Network Activity ◽

Neuronal Cultures ◽

Primary Neuronal Cultures

Download Full-text

Difluoroboron β-diketonate polylactic acid oxygen nanosensors for intracellular neuronal imaging

Scientific Reports ◽

10.1038/s41598-020-80172-w ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Meng Zhuang ◽

Suchitra Joshi ◽

Huayu Sun ◽

Tamal Batabyal ◽

Cassandra L. Fraser ◽

...

Keyword(s):

Neuronal Activity ◽

Oxygen Sensing ◽

Brain Slices ◽

Neuronal Cell ◽

Quantum Yields ◽

Poly Lactic Acid ◽

Neuronal Cultures ◽

Primary Neuronal Cultures ◽

Non Invasive ◽

Mitotracker Red

AbstractCritical for metabolism, oxygen plays an essential role in maintaining the structure and function of neurons. Oxygen sensing is important in common neurological disorders such as strokes, seizures, or neonatal hypoxic–ischemic injuries, which result from an imbalance between metabolic demand and oxygen supply. Phosphorescence quenching by oxygen provides a non-invasive optical method to measure oxygen levels within cells and tissues. Difluoroboron β-diketonates are a family of luminophores with high quantum yields and tunable fluorescence and phosphorescence when embedded in certain rigid matrices such as poly (lactic acid) (PLA). Boron nanoparticles (BNPs) can be fabricated from dye-PLA materials for oxygen mapping in a variety of biological milieu. These dual-emissive nanoparticles have oxygen-insensitive fluorescence, oxygen-sensitive phosphorescence, and rigid matrix all in one, enabling real-time ratiometric oxygen sensing at micron-level spatial and millisecond-level temporal resolution. In this study, BNPs are applied in mouse brain slices to investigate oxygen distributions and neuronal activity. The optical properties and physical stability of BNPs in a biologically relevant buffer were stable. Primary neuronal cultures were labeled by BNPs and the mitochondria membrane probe MitoTracker Red FM. BNPs were taken up by neuronal cell bodies, at dendrites, and at synapses, and the localization of BNPs was consistent with that of MitoTracker Red FM. The brain slices were stained with the BNPs, and the BNPs did not significantly affect the electrophysiological properties of neurons. Oxygen maps were generated in living brain slices where oxygen is found to be mostly consumed by mitochondria near synapses. Finally, the BNPs exhibited excellent response when the conditions varied from normoxic to hypoxic and when the neuronal activity was increased by increasing K+ concentration. This work demonstrates the capability of BNPs as a non-invasive tool in oxygen sensing and could provide fundamental insight into neuronal mechanisms and excitability research.

Download Full-text

Ferroptosis is controlled by the coordinated transcriptional regulation of glutathione and labile iron metabolism by the transcription factor BACH1

Journal of Biological Chemistry ◽

10.1074/jbc.ra119.009548 ◽

2019 ◽

Vol 295 (1) ◽

pp. 69-82 ◽

Cited By ~ 18

Author(s):

Hironari Nishizawa ◽

Mitsuyo Matsumoto ◽

Tomohiko Shindo ◽

Daisuke Saigusa ◽

Hiroki Kato ◽

...

Keyword(s):

Transcription Factor ◽

Transcriptional Regulation ◽

Iron Metabolism ◽

Labile Iron

Download Full-text

Integrated transcription factor profiling with transcriptome analysis identifies L1PA2 transposons as global regulatory modulators in a breast cancer model

Scientific Reports ◽

10.1038/s41598-021-86395-9 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Jiayue-Clara Jiang ◽

Joseph A. Rothnagel ◽

Kyle R. Upton

Keyword(s):

Breast Cancer ◽

Transcription Factor ◽

Transcriptional Regulation ◽

Disease Diagnosis ◽

Integrated Analysis ◽

Regulatory Sequences ◽

Cancer Model ◽

Mcf7 Cells ◽

Epithelial Cancers ◽

Breast Cancer Model

AbstractWhile transposons are generally silenced in somatic tissues, many transposons escape epigenetic repression in epithelial cancers, become transcriptionally active and contribute to the regulation of human gene expression. We have developed a bioinformatic pipeline for the integrated analysis of transcription factor binding and transcriptomic data to identify transposon-derived promoters that are activated in specific diseases and developmental states. We applied this pipeline to a breast cancer model, and found that the L1PA2 transposon subfamily contributes abundant regulatory sequences to co-ordinated transcriptional regulation in breast cancer. Transcription factor profiling demonstrates that over 27% of L1PA2 transposons harbour co-localised binding sites of functionally interacting, cancer-associated transcription factors in MCF7 cells, a cell line used to model breast cancer. Transcriptomic analysis reveals that L1PA2 transposons also contribute transcription start sites to up-regulated transcripts in MCF7 cells, including some transcripts with established oncogenic properties. In addition, we verified the utility of our pipeline on other transposon subfamilies, as well as on leukemia and lung carcinoma cell lines. We demonstrate that the normally quiescent regulatory activities of transposons can be activated and alter the cancer transcriptome. In particular, the L1PA2 subfamily contributes abundant regulatory sequences, and likely plays a global role in modulating breast cancer transcriptional regulation. Understanding the regulatory impact of L1PA2 on breast cancer genomes provides additional insights into cancer genome regulation, and may provide novel biomarkers for disease diagnosis, prognosis and therapy.

Download Full-text

Oxidative Stress Induces Dephosphorylation of τ in Rat Brain Primary Neuronal Cultures

Journal of Neurochemistry ◽

10.1046/j.1471-4159.1997.68041590.x ◽

2002 ◽

Vol 68 (4) ◽

pp. 1590-1597 ◽

Cited By ~ 30

Author(s):

Daniel R. Davis ◽

Brian H. Anderton ◽

Jean-Pierre Brion ◽

C. Hugh Reynolds ◽

Diane P. Hanger

Keyword(s):

Oxidative Stress ◽

Rat Brain ◽

Neuronal Cultures ◽

Primary Neuronal Cultures

Download Full-text

Ketamine decreases neuronally released glutamate via retrograde stimulation of presynaptic adenosine A1 receptors

Molecular Psychiatry ◽

10.1038/s41380-021-01246-3 ◽

2021 ◽

Author(s):

Vesna Lazarevic ◽

Yunting Yang ◽

Ivana Flais ◽

Per Svenningsson

Keyword(s):

Cortical Neurons ◽

Intracellular Signaling ◽

Ex Vivo ◽

Forced Swim Test ◽

Glutamate Release ◽

Neuronal Cultures ◽

Extracellular Glutamate ◽

Primary Neuronal Cultures ◽

Adenosine A1 Receptors ◽

Adenosine A1

AbstractKetamine produces a rapid antidepressant response in patients with major depressive disorder (MDD), but the underlying mechanisms appear multifaceted. One hypothesis, proposes that by antagonizing NMDA receptors on GABAergic interneurons, ketamine disinhibits afferens to glutamatergic principal neurons and increases extracellular glutamate levels. However, ketamine seems also to reduce rapid glutamate release at some synapses. Therefore, clinical studies in MDD patients have stressed the need to identify mechanisms whereby ketamine decreases presynaptic activity and glutamate release. In the present study, the effect of ketamine and its antidepressant metabolite, (2R,6R)-HNK, on neuronally derived glutamate release was examined in rodents. We used FAST methodology to measure depolarization-evoked extracellular glutamate levels in vivo in freely moving or anesthetized animals, synaptosomes to detect synaptic recycling ex vivo and primary cortical neurons to perform functional imaging and to examine intracellular signaling in vitro. In all these versatile approaches, ketamine and (2R,6R)-HNK reduced glutamate release in a manner which could be blocked by AMPA receptor antagonism. Antagonism of adenosine A1 receptors, which are almost exclusively expressed at nerve terminals, also counteracted ketamine’s effect on glutamate release and presynaptic activity. Signal transduction studies in primary neuronal cultures demonstrated that ketamine reduced P-T286-CamKII and P-S9-Synapsin, which correlated with decreased synaptic vesicle recycling. Moreover, systemic administration of A1R antagonist counteracted the antidepressant-like actions of ketamine and (2R,6R)-HNK in the forced swim test. To conclude, by studying neuronally released glutamate, we identified a novel retrograde adenosinergic feedback mechanism that mediate inhibitory actions of ketamine on glutamate release that may contribute to its rapid antidepressant action.

Download Full-text

Role of Ca2+ in α-Amino-3-Hydroxy-5-Methyl-4-Isoxazolepropionic Acid-Mediated Polyphosphoinositide Turnover in Primary Neuronal Cultures

Journal of Neurochemistry ◽

10.1046/j.1471-4159.1994.62062325.x ◽

2008 ◽

Vol 62 (6) ◽

pp. 2325-2332 ◽

Cited By ~ 14

Author(s):

Yi-Hsuan Lee ◽

David L. Deupree ◽

Shine-Chi Chen ◽

Lung-Sen Kao ◽

Jang-Yen Wu

Keyword(s):

Neuronal Cultures ◽

Primary Neuronal Cultures

Download Full-text

Regulation of Cholesterol Homeostasis by the Liver X Receptors in the Central Nervous System

Molecular Endocrinology ◽

10.1210/mend.16.6.0835 ◽

2002 ◽

Vol 16 (6) ◽

pp. 1378-1385 ◽

Cited By ~ 78

Author(s):

Karl D. Whitney ◽

Michael A. Watson ◽

Jon L. Collins ◽

William G. Benson ◽

Tammy M. Stone ◽

...

Keyword(s):

Gene Expression ◽

Central Nervous System ◽

Nervous System ◽

Cholesterol Efflux ◽

Target Genes ◽

Cholesterol Homeostasis ◽

Neuronal Cultures ◽

Primary Neuronal Cultures ◽

The Central Nervous System ◽

Lxr Agonists

Abstract The nuclear oxysterol receptors liver X receptor-α [LXRα (NR1H3)] and LXRβ (NR1H2) coordinately regulate genes involved in cholesterol homeostasis. Although both LXR subtypes are expressed in the brain, their roles in this tissue remain largely unexplored. In this report, we show that LXR agonists have marked effects on gene expression in murine brain tissue both in vitro and in vivo. In primary astrocyte cultures, LXR agonists regulated several established LXR target genes, including ATP binding cassette transporter A1, and enhanced cholesterol efflux. In contrast, little or no effect on gene expression or cholesterol efflux was detected in primary neuronal cultures. Treatment of mice with a selective LXR agonist resulted in the induction of several LXR target genes related to cholesterol homeostasis in the cerebellum and hippocampus. These data provide the first evidence that the LXRs regulate cholesterol homeostasis in the central nervous system. Because dysregulation of cholesterol balance is implicated in central nervous system diseases such as Alzheimer’s and Niemann-Pick disease, pharmacological manipulation of the LXRs may prove beneficial in the treatment of these disorders.

Download Full-text