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Amin Sherafat, Robert A Hill, Akiko Nishiyama
Oligodendrocyte development and myelination are processes in the central nervous system that are regulated by cell intrinsic and extrinsic mechanisms. Organotypic slice cultures provide a simple method for studying factors that affect oligodendrocyte proliferation, differentiation, and myelination in the context of the local cellular environment. Here we show that major glial cell types and neurons are preserved in slice cultures from postnatal mouse forebrain, and their morphological characteristics are retained...
2018: Methods in Molecular Biology
Sooyoung Cho, Yeon Joo Jung, Eun Cheng Suh, Hee Jung Baik, Jong In Han, Guie Yong Lee, Kyung Eun Lee
Propofol is the most widely accepted intravenous anesthetic available for clinical use. However, neurotoxicity of propofol in the developing brain has been reported. This study investigated the effects of propofol on cognitive function in normal healthy adult mice. Thirty-three GFP-LC3 adult mice were included. Propofol was injected for anesthesia (n= 22). The sham control (n= 11) received intralipid injections. The mice completed a Y-maze test on 3 and 7 days after being anesthetized. Western blotting, immunofluorescence staining, and transmission electron microscopic (TEM) analyses were performed with their hippocampi...
July 10, 2018: Brain Research
Fabian A Grimm, Alexander Blanchette, John S House, Kyle Ferguson, Nan-Hung Hsieh, Chimeddulam Dalaijamts, Alec A Wright, Blake Anson, Fred A Wright, Weihsueh A Chiu, Ivan Rusyn
Assessing inter-individual variability in responses to xenobiotics remains a substantial challenge, both in drug development with respect to pharmaceuticals and in public health with respect to environmental chemicals.  Although approaches exist to characterize pharmacokinetic variability, there are no methods to routinely address pharmacodynamic variability.  In this study, we aimed to demonstrate the feasibility of characterizing inter-individual variability in a human in vitro model. Specifically, we hypothesized that genetic variability across a population of iPSC-derived cardiomyocytes translates into reproducible variability in both baseline phenotypes and drug responses...
July 8, 2018: ALTEX
Olga Ilina, Leonard Campanello, Pavel G Gritsenko, Manon Vullings, Chenlu Wang, Peter Bult, Wolfgang Losert, Peter Friedl
Cancer invasion programs are adaptive by switching between metastatic collective and single-cell dissemination, however current intravital microscopy models for epithelial cancer in mice fail to reliably recreate such invasion plasticity. Using image-guided microimplantation of breast cancer spheroids into the murine mammary fat pad and live-cell monitoring, we show microenvironmental conditions and cytoskeletal adaptation during collective to single-cell transition in vivo E-cadherin expressing 4T1 and E-cadherin negative MMT tumors both initiated collective invasion along stromal structures, reflecting invasion patterns in 3D organotypic culture and human primary ductal and lobular carcinoma...
July 11, 2018: Disease Models & Mechanisms
Daniela M Magalhães, Noémia Pereira, Diogo M Rombo, Cláudia Beltrão-Cavacas, Ana M Sebastião, Cláudia A Valente
BACKGROUND: Epilepsy is a prevalent neurological disorder worldwide. It is characterized by an enduring predisposition to generate seizures and its development is accompanied by alterations in many cellular processes. Organotypic slice cultures represent a multicellular environment with the potential to assess biological mechanisms, and they are used as a starting point for refining molecules for in vivo studies. Here, we investigated organotypic slice cultures as a model of epilepsy...
July 11, 2018: Journal of Neuroinflammation
Pavlo G Gritsenko, Peter Friedl
Diffuse brain invasion by glioma cells prevents effective surgical or molecular-targeted therapy and underlies detrimental outcome. Migrating glioma cells are guided by complex anatomical brain structures, however exact mechanisms remain poorly defined. To identify adhesion receptor systems and matrix structures supporting glioma cell invasion into brain-like environments we used 2D and 3D organotypic invasion assays in combination with antibody-, peptide- and RNA-based interference. Combined interference with β1 and αV integrins abolished the migration of U-251 and E-98 glioma cells on reconstituted basement membrane, however invasion into primary brain slices or 3D astrocyte-based scaffolds and migration on astrocyte-deposited matrix was only partly inhibited...
July 10, 2018: Journal of Cell Science
Chad Curtis, Dorsa Toghani, Ben Wong, Elizabeth Nance
Drug delivery to the brain is challenging due to a highly regulated blood-brain barrier (BBB) and a complex brain microenvironment. Nanoparticles, due to their tailorability, provide promising platforms to enhance therapeutic delivery and achieve controlled release and disease-specific localization in the brain. However, we have yet to fully understand the complex interactions between nanoparticles and the biological environments in which they operate. It is important to perform a systematic study to characterize nanoparticle behavior as a function of ion composition, concentration, and pH in cerebrospinal fluid (CSF)...
June 30, 2018: Colloids and Surfaces. B, Biointerfaces
Marta Figueiredo, Hélia Neves
The avian embryo, as an experimental model, has been of utmost importance for seminal discoveries in developmental biology. Among several approaches, the formation of quail-chicken chimeras and the use of the chorioallantoic membrane (CAM) to sustain the development of ectopic tissues date back to the last century. Nowadays, the combination of these classical techniques with recent in vitro methodologies offers novel prospects to further explore organ formation. Here we describe a two-step approach to study early- and late-stages of organogenesis...
June 17, 2018: Journal of Visualized Experiments: JoVE
Tatiana A Karakasheva, Eric W Lin, Qiaosi Tang, Edmund Qiao, Todd J Waldron, Monica Soni, Andres J Klein-Szanto, Varun Sahu, Devraj Basu, Shinya Ohashi, Kiichiro Baba, Zachary T Giaccone, Sarah R Walker, David A Frank, E Paul Wileyto, Qi Long, Margaret Dunagin, Arjun Raj, J Alan Diehl, Kwok-Kin Wong, Adam J Bass, Anil K Rustgi
The tumor microenvironment (TME) plays a major role in the pathogenesis of multiple cancer types, including upper-gastrointestinal (GI) cancers that currently lack effective therapeutic options. Cancer-associated fibroblasts (CAF) are an essential component of the TME, contributing to tumorigenesis by secreting growth factors, modifying the extracellular matrix, supporting angiogenesis, and suppressing anti-tumor immune responses. Through an unbiased approach, we have established that IL-6 mediates crosstalk between tumor cells and CAF not only by supporting tumor cell growth, but also by promoting fibroblast activation...
July 5, 2018: Cancer Research
Yun Qiao, Quan Dong, Baichen Li, Sofian Obaid, Christian Miccile, Rose T Yin, Trisha Talapatra, Zexu Lin, Sihui Li, Zhenyu Li, Igor R Efimov
Human cardiac slices have emerged as a promising model of the human heart for scientific research and drug testing. Retaining the normal tissue architecture, a multi-cell type environment, and the native extracellular matrix, human cardiac slices faithfully replicate organ-level adult cardiac physiology. Previously, we demonstrated that human cardiac tissue slices cultured for 24 h maintained normal electrophysiology. In this project, we further optimized the organotypic culture condition to maintain normal electrophysiology of the human cardiac slices for 4 days...
June 27, 2018: Progress in Biophysics and Molecular Biology
Elisa Landucci, Irene L Llorente, Berta Anuncibay-Soto, Domenico E Pellegrini-Giampietro, Arsenio Fernández-López
We previously demonstrated that the non-steroidal anti-inflammatory agent meloxicam has neuroprotective effects in an oxygen and glucose deprivation model (OGD) of rat organotypic hippocampal slice cultures. We wondered if GABAergic transmission changed the neuroprotective effects of meloxicam and if meloxicam was able to modulate endoplasmic reticulum stress (ER stress) in this model. Mortality was measured using propidium iodide. Western blot assays were performed to measure levels of cleaved and non-cleaved caspase-3 to quantify apoptosis, while levels of GRP78, GRP94 and phosphorylated eIF2α were used to detect unfolded protein response (UPR)...
June 24, 2018: Neuroscience
Charlotte Rodrigues Neves, Susan Gibbs
Contact with the skin is inevitable or desirable for daily life products such as cosmetics, hair dyes, perfumes, drugs, household products, and industrial and agricultural products. Whereas the majority of these products are harmless, a number can become metabolized and/or activate the immunological defense via innate and adaptive mechanisms resulting in sensitization and allergic contact dermatitis upon following exposures to the same substance. Therefore, strict safety (hazard) assessment of actives and ingredients in products and drugs applied to the skin is essential to determine I) whether the chemical is a potential sensitizer and if so II) what is the safe concentration for human exposure to prevent sensitization from occurring...
June 23, 2018: Current Topics in Microbiology and Immunology
Michael H Hastings, Elizabeth S Maywood, Marco Brancaccio
The suprachiasmatic nucleus (SCN) of the hypothalamus is remarkable. Despite numbering only about 10,000 neurons on each side of the third ventricle, the SCN is our principal circadian clock, directing the daily cycles of behaviour and physiology that set the tempo of our lives. When this nucleus is isolated in organotypic culture, its autonomous timing mechanism can persist indefinitely, with precision and robustness. The discovery of the cell-autonomous transcriptional and post-translational feedback loops that drive circadian activity in the SCN provided a powerful exemplar of the genetic specification of complex mammalian behaviours...
June 22, 2018: Nature Reviews. Neuroscience
Noémie Remacle, Patrick Forny, Hong-Phuc Cudré-Cung, Mary Gonzalez-Melo, Sónia do Vale-Pereira, Hugues Henry, Tony Teav, Hector Gallart-Ayala, Olivier Braissant, Matthias Baumgartner, Diana Ballhausen
BACKGROUND: Methylmalonic aciduria (MMAuria) is an inborn error of metabolism leading to neurological deterioration. In this study, we used 3D organotypic brain cell cultures derived from embryos of a brain-specific Mut-/- (brain KO) mouse to investigate mechanisms leading to brain damage. We challenged our in vitro model by a catabolic stress (temperature shift). RESULTS: Typical metabolites for MMAuria as well as a massive NH4+ increase were found in the media of brain KO cultures...
June 18, 2018: Molecular Genetics and Metabolism
Nina M Poole, Anubama Rajan, Anthony W Maresso
Adherence, invasion, and translocation to and through the intestinal epithelium are important drivers of disease for many enteric bacteria. However, most work has been limited to transformed intestinal cell lines or murine models that often do not faithfully recapitulate key elements associated with human disease. The recent technological advances in organotypic tissue and cell culture are providing unparalleled access to systems with human physiology and complexity. Human intestinal enteroids (HIEs), derived from patient biopsy or surgical specimens of intestinal tissues, are organotypic cultures now being adapted to the study of enteric infections...
May 17, 2018: Current Protocols in Microbiology
Duo Xu, Shun-Qing Liang, Haitang Yang, Ursina Lüthi, Carsten Riether, Sabina Berezowska, Thomas M Marti, Sean R R Hall, Rémy Bruggmann, Gregor J Kocher, Ralph A Schmid, Ren-Wang Peng
BACKGROUND: Standard treatment for advanced malignant pleural mesothelioma (MPM) is a cisplatin/pemetrexed (MTA) regimen; however, this is confronted by drug resistance. Proteotoxic stress in the endoplasmic reticulum (ER) is a hallmark of cancer and some rely on this stress signalling in response to cytotoxic chemotherapeutics. We hypothesise that ER stress and the adaptive unfolded protein response (UPR) play a role in chemotherapy resistance of MPM. METHODS: In vitro three-dimensional (3D) and ex vivo organotypic culture were used to enrich a chemotherapy-resistant population and recapitulate an in vivo MPM microenvironment, respectively...
June 20, 2018: British Journal of Cancer
Ksenia Gnedeva, A J Hudspeth, Neil Segil
The sensory organs of the inner ear are challenging to study in mammals due to their inaccessibility to experimental manipulation and optical observation. Moreover, although existing culture techniques allow biochemical perturbations, these methods do not provide a means to study the effects of mechanical force and tissue stiffness during development of the inner ear sensory organs. Here we describe a method for three-dimensional organotypic culture of the intact murine utricle and cochlea that overcomes these limitations...
June 1, 2018: Journal of Visualized Experiments: JoVE
Cara L Croft, Wendy Noble
Alzheimer's disease, the most common cause of dementia, is a progressive neurodegenerative disorder characterised by amyloid-beta deposits in extracellular plaques, intracellular neurofibrillary tangles of aggregated tau, synaptic dysfunction and neuronal death. There are no cures for AD and current medications only alleviate some disease symptoms. Transgenic rodent models to study Alzheimer's mimic features of human disease such as age-dependent accumulation of abnormal beta-amyloid and tau, synaptic dysfunction, cognitive deficits and neurodegeneration...
2018: F1000Research
ArunSundar MohanaSundaram, Thukani Sathanantham Shanmugarajan, Velayutham Ravichandiran
Cognitive impulsivity, a form of suboptimal cost-benefit decision making is an illustrious attribute of an array of neurodegenerative diseases including Alzheimer's Disease (AD). In this study, delay discounting paradigm was used to assess the effect of 3,4-dihydroxyphenylethanol (DOPET) on cognitive impulsivity-in an oA42i (oligomeric amyloid β1-42 plus ibotenic acid) induced AD mouse model-using a non-spatial T-maze task. The results depicted that oA42i administration elevated cognitive impulsivity, whereas DOPET treatment attenuated the impulsive behavior and matched the choice of the sham-operated controls...
June 14, 2018: ACS Chemical Neuroscience
Alissa Piekarski, Gurueswar Nagarajan, Peter Ishola, Joshua Flees, Elizabeth S Greene, Wayne J Kuenzel, Takeshi Ohkubo, Helena Maier, Walter G Bottje, Mark A Cline, Sami Dridi
Autophagy, a highly conserved intracellular self-digestion process, plays an integral role in maintaining cellular homeostasis. Although emerging evidence indicate that the endocrine system regulates autophagy in mammals, there is still a scarcity of information on autophagy in avian (non-mammalian) species. Here, we show that intracerebroventricular administration of leptin reduces feed intake, modulates the expression of feeding-related hypothalamic neuropeptides, activates leptin receptor and signal transducer and activator of transcription (Ob-Rb/STAT) pathway, and significantly increases the expression of autophagy-related proteins (Atg3, Atg5, Atg7, beclin1, and LC3B) in chicken hypothalamus, liver, and muscle...
2018: Frontiers in Physiology
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