133 results on '"Per Uhlén"'
Search Results
2. Human ex vivo spinal cord slice culture as a useful model of neural development, lesion, and allogeneic neural cell therapy
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Chenhong Lin, Cinzia Calzarossa, Teresa Fernandez-Zafra, Jia Liu, Xiaofei Li, Åsa Ekblad-Nordberg, Erika Vazquez-Juarez, Simone Codeluppi, Lena Holmberg, Maria Lindskog, Per Uhlén, and Elisabet Åkesson
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Spinal cord injury ,Stem cell therapy ,Human organotypic culture ,Medicine (General) ,R5-920 ,Biochemistry ,QD415-436 - Abstract
Abstract Background There are multiple promising treatment strategies for central nervous system trauma and disease. However, to develop clinically potent and safe treatments, models of human-specific conditions are needed to complement in vitro and in vivo animal model-based studies. Methods We established human brain stem and spinal cord (cross- and longitudinal sections) organotypic cultures (hOCs) from first trimester tissues after informed consent by donor and ethical approval by the Regional Human Ethics Committee, Stockholm (lately referred to as Swedish Ethical Review Authority), and The National Board of Health and Welfare, Sweden. We evaluated the stability of hOCs with a semi-quantitative hOC score, immunohistochemistry, flow cytometry, Ca2+ signaling, and electrophysiological analysis. We also applied experimental allogeneic human neural cell therapy after injury in the ex vivo spinal cord slices. Results The spinal cord hOCs presented relatively stable features during 7–21 days in vitro (DIV) (except a slightly increased cell proliferation and activated glial response). After contusion injury performed at 7 DIV, a significant reduction of the hOC score, increase of the activated caspase-3+ cell population, and activated microglial populations at 14 days postinjury compared to sham controls were observed. Such elevation in the activated caspase-3+ population and activated microglial population was not observed after allogeneic human neural cell therapy. Conclusions We conclude that human spinal cord slice cultures have potential for future structural and functional studies of human spinal cord development, injury, and treatment strategies.
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- 2020
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3. BCG‐induced cytokine release in bladder cancer cells is regulated by Ca2+ signaling
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Cristián Ibarra, Marie Karlsson, Simone Codeluppi, Manuel Varas‐Godoy, Songbai Zhang, Lauri Louhivuori, Sara Mangsbo, Arad Hosseini, Navid Soltani, Rahim Kaba, T. Kalle Lundgren, Abolfazl Hosseini, Nobuyuki Tanaka, Mototsugu Oya, Peter Wiklund, Ayako Miyakawa, and Per Uhlén
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BCG ,calcium signaling ,TLR4 ,urinary bladder cancer ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Bacillus Calmette–Guérin (BCG) is widely used in the clinic to effectively treat superficial urinary bladder cancer. However, a significant proportion of patients who fail to respond to BCG risk cystectomy or death. Though more than 3 million cancer treatments with BCG occur annually, surprisingly little is known about the initial signaling cascades activated by BCG. Here, we report that BCG induces a rapid intracellular Ca2+ (calcium ion) signal in bladder cancer cells that is essential for activating the transcription factor nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB) and for synthesizing and secreting proinflammatory cytokines, including interleukin 8 (IL‐8). A similar Ca2+ response was observed when cells were exposed to the supernatant of BCG. Studying cellular molecular mechanisms involved in the BCG signaling event, we found pivotal roles for phospholipase C and the Toll‐like receptor 4. Further assessment revealed that this signaling pathway induces synthesis of IL‐8, whereas exocytosis appeared to be controlled by global Ca2+ signaling. These results shed new light on the molecular mechanisms underlying BCG treatment of bladder cancer, which can help in improving therapeutic efficacy and reducing adverse side effects.
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- 2019
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4. Recombinant Spider Silk Protein Matrices Facilitate Differentiation of Neural Stem Cells Into Mature and Functional Neurons
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Michalina Lewicka, Paola Rebellato, Jakub Lewicki, Per Uhlén, Anna Rising, and Ola Hermanson
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neural stem cell ,biomaterial ,spider silk ,3D cultures ,scaffold ,Technology - Abstract
Neural stem cells (NSCs) show great promise in drug discovery and clinical application. Yet few efforts have been made to optimize biocompatible materials for such cells to be expanded and used in clinical conditions. We have previously demonstrated that NSCs are readily cultured on substrates of certain recombinant spider silk protein without addition of animal- or human-derived components. The question remains however whether this material allows differentiation into functional neurons, and whether such differentiation can take place also when the NSCs are cultured not only upon but also within the biodegradable material. Here we demonstrate that “foam”-like structures generated from recombinant spider silk protein (4RepCT) provided excellent matrices for the generation and multicellular analysis of functional excitatory neurons from NSCs without addition of animal- or human-derived components. NSCs isolated from the cerebral cortices of rat embryos were cultured at either 4RepCT matrices shaped as foam-like structures without coating, or on conventional polystyrene plates coated with poly-L-ornithine and fibronectin. Upon treatment with recombinant proteins including the extracellular signaling factor BMP4 or a combination of BMP4 and the signaling factor Wnt3a, the cortical NSCs cultured in 4RepCT foam-like structures differentiated efficiently into neurons that responded to glutamate receptor agonists, such as AMPA, to the same extent as control cultures. Matrices derived from recombinant spider silk proteins thus provide a functional microenvironment for neural stem cells with little or no animal- or human-derived components and can be employed in the development of new strategies in stem cell research and tissue engineering.
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- 2021
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5. Radiation Triggers a Dynamic Sequence of Transient Microglial Alterations in Juvenile Brain
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Ahmed M. Osman, Ying Sun, Terry C. Burns, Liqun He, Nigel Kee, Nuria Oliva-Vilarnau, Androniki Alevyzaki, Kai Zhou, Lauri Louhivuori, Per Uhlén, Eva Hedlund, Christer Betsholtz, Volker M. Lauschke, Julianna Kele, and Klas Blomgren
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radiotherapy ,neuroinflammation ,M1 and M2 ,transcriptome ,RNA sequencing ,Biology (General) ,QH301-705.5 - Abstract
Summary: Cranial irradiation (IR), an effective tool to treat malignant brain tumors, triggers a chronic pro-inflammatory microglial response, at least in the adult brain. Using single-cell and bulk RNA sequencing, combined with histology, we show that the microglial response in the juvenile mouse hippocampus is rapid but returns toward normal within 1 week. The response is characterized by a series of temporally distinct homeostasis-, sensome-, and inflammation-related molecular signatures. We find that a single microglial cell simultaneously upregulates transcripts associated with pro- and anti-inflammatory microglial phenotypes. Finally, we show that juvenile and adult irradiated microglia are already transcriptionally distinct in the early phase after IR. Our results indicate that microglia are involved in the initial stages but may not be responsible for driving long-term inflammation in the juvenile brain.
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- 2020
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6. Retraction: Ghali MGZ, et al. Mechanisms underlying the generation of autonomorespiratory coupling amongst the respiratory central pattern generator, sympathetic oscillators, and cardiovagal premotoneurons. Journal of Integrative Neuroscience. 2020; 19: 521–560
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Michael G. Z. Ghali, George Zaki Ghali, Adriana Lima, Michael McDermott, Emma Glover, Stefanos Voglis, Jennifer Humphrey, Marton Skog Steinberger König, Henry Brem, Per Uhlén, Robert F. Spetzler, M. Gazi Yasargil, and Journal of Integrative Neuroscience Editorial Office
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Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Abstract
No abstract present.
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- 2021
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7. Wnt/β-Catenin Stimulation and Laminins Support Cardiovascular Cell Progenitor Expansion from Human Fetal Cardiac Mesenchymal Stromal Cells
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Agneta Månsson-Broberg, Sergey Rodin, Ivana Bulatovic, Cristián Ibarra, Marie Löfling, Rami Genead, Eva Wärdell, Ulrika Felldin, Carl Granath, Evren Alici, Katarina Le Blanc, C.I. Edvard Smith, Alena Salašová, Magnus Westgren, Erik Sundström, Per Uhlén, Ernest Arenas, Christer Sylvén, Karl Tryggvason, Matthias Corbascio, Oscar E. Simonson, Cecilia Österholm, and Karl-Henrik Grinnemo
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Medicine (General) ,R5-920 ,Biology (General) ,QH301-705.5 - Abstract
The intrinsic regenerative capacity of human fetal cardiac mesenchymal stromal cells (MSCs) has not been fully characterized. Here we demonstrate that we can expand cells with characteristics of cardiovascular progenitor cells from the MSC population of human fetal hearts. Cells cultured on cardiac muscle laminin (LN)-based substrata in combination with stimulation of the canonical Wnt/β-catenin pathway showed increased gene expression of ISL1, OCT4, KDR, and NKX2.5. The majority of cells stained positive for PDGFR-α, ISL1, and NKX2.5, and subpopulations also expressed the progenitor markers TBX18, KDR, c-KIT, and SSEA-1. Upon culture of the cardiac MSCs in differentiation media and on relevant LNs, portions of the cells differentiated into spontaneously beating cardiomyocytes, and endothelial and smooth muscle-like cells. Our protocol for large-scale culture of human fetal cardiac MSCs enables future exploration of the regenerative functions of these cells in the context of myocardial injury in vitro and in vivo.
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- 2016
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8. Correction to: Human ex vivo spinal cord slice culture as a useful model of neural development, lesion, and allogeneic neural cell therapy
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Chenhong Lin, Cinzia Calzarossa, Teresa Fernandez-Zafra, Jia Liu, Xiaofei Li, Åsa Ekblad-Nordberg, Erika Vazquez-Juarez, Simone Codeluppi, Lena Holmberg, Maria Lindskog, Per Uhlén, and Elisabet Åkesson
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Medicine (General) ,R5-920 ,Biochemistry ,QD415-436 - Abstract
An amendment to this paper has been published and can be accessed via the original article.
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- 2020
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9. Voltage-dependent calcium channel signaling mediates GABAA receptor-induced migratory activation of dendritic cells infected by Toxoplasma gondii.
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Sachie Kanatani, Jonas M Fuks, Einar B Olafsson, Linda Westermark, Benedict Chambers, Manuel Varas-Godoy, Per Uhlén, and Antonio Barragan
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
The obligate intracellular parasite Toxoplasma gondii exploits cells of the immune system to disseminate. Upon T. gondii-infection, γ-aminobutyric acid (GABA)/GABAA receptor signaling triggers a hypermigratory phenotype in dendritic cells (DCs) by unknown signal transduction pathways. Here, we demonstrate that calcium (Ca2+) signaling in DCs is indispensable for T. gondii-induced DC hypermotility and transmigration in vitro. We report that activation of GABAA receptors by GABA induces transient Ca2+ entry in DCs. Murine bone marrow-derived DCs preferentially expressed the L-type voltage-dependent Ca2+ channel (VDCC) subtype Cav1.3. Silencing of Cav1.3 by short hairpin RNA or selective pharmacological antagonism of VDCCs abolished the Toxoplasma-induced hypermigratory phenotype. In a mouse model of toxoplasmosis, VDCC inhibition of adoptively transferred Toxoplasma-infected DCs delayed the appearance of cell-associated parasites in the blood circulation and reduced parasite dissemination to target organs. The present data establish that T. gondii-induced hypermigration of DCs requires signaling via VDCCs and that Ca2+ acts as a second messenger to GABAergic signaling via the VDCC Cav1.3. The findings define a novel motility-related signaling axis in DCs and unveil that interneurons and DCs share common GABAergic motogenic pathways. T. gondii employs GABAergic non-canonical pathways to induce host cell migration and facilitate dissemination.
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- 2017
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10. CO2-evoked release of PGE2 modulates sighs and inspiration as demonstrated in brainstem organotypic culture
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David Forsberg, Zachi Horn, Evangelia Tserga, Erik Smedler, Gilad Silberberg, Yuri Shvarev, Kai Kaila, Per Uhlén, and Eric Herlenius
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respiration ,prostaglandins ,neural network ,chemosensitivity ,calcium imaging ,small world ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Inflammation-induced release of prostaglandin E2 (PGE2) changes breathing patterns and the response to CO2 levels. This may have fatal consequences in newborn babies and result in sudden infant death. To elucidate the underlying mechanisms, we present a novel breathing brainstem organotypic culture that generates rhythmic neural network and motor activity for 3 weeks. We show that increased CO2 elicits a gap junction-dependent release of PGE2. This alters neural network activity in the preBötzinger rhythm-generating complex and in the chemosensitive brainstem respiratory regions, thereby increasing sigh frequency and the depth of inspiration. We used mice lacking eicosanoid prostanoid 3 receptors (EP3R), breathing brainstem organotypic slices and optogenetic inhibition of EP3R+/+ cells to demonstrate that the EP3R is important for the ventilatory response to hypercapnia. Our study identifies a novel pathway linking the inflammatory and respiratory systems, with implications for inspiration and sighs throughout life, and the ability to autoresuscitate when breathing fails.
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- 2016
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11. Sublethal caspase activation promotes generation of cardiomyocytes from embryonic stem cells.
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Ivana Bulatovic, Cristian Ibarra, Cecilia Österholm, Heng Wang, Antonio Beltrán-Rodríguez, Manuel Varas-Godoy, Agneta Månsson-Broberg, Per Uhlén, András Simon, and Karl-Henrik Grinnemo
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Medicine ,Science - Abstract
Generation of new cardiomyocytes is critical for cardiac repair following myocardial injury, but which kind of stimuli is most important for cardiomyocyte regeneration is still unclear. Here we explore if apoptotic stimuli, manifested through caspase activation, influences cardiac progenitor up-regulation and cardiomyocyte differentiation. Using mouse embryonic stem cells as a cellular model, we show that sublethal activation of caspases increases the yield of cardiomyocytes while concurrently promoting the proliferation and differentiation of c-Kit+/α-actininlow cardiac progenitor cells. A broad-spectrum caspase inhibitor blocked these effects. In addition, the caspase inhibitor reversed the mRNA expression of genes expressed in cardiomyocytes and their precursors. Our study demonstrates that sublethal caspase-activation has an important role in cardiomyocyte differentiation and may have significant implications for promoting cardiac regeneration after myocardial injury involving exogenous or endogenous cell sources.
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- 2015
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12. Infection by Toxoplasma gondii Induces Amoeboid-Like Migration of Dendritic Cells in a Three-Dimensional Collagen Matrix.
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Sachie Kanatani, Per Uhlén, and Antonio Barragan
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Medicine ,Science - Abstract
Toxoplasma gondii, an obligate intracellular parasite of humans and other warm-blooded vertebrates, invades a variety of cell types in the organism, including immune cells. Notably, dendritic cells (DCs) infected by T. gondii acquire a hypermigratory phenotype that potentiates parasite dissemination by a 'Trojan horse' type of mechanism in mice. Previous studies have demonstrated that, shortly after parasite invasion, infected DCs exhibit hypermotility in 2-dimensional confinements in vitro and enhanced transmigration in transwell systems. However, interstitial migration in vivo involves interactions with the extracellular matrix in a 3-dimensional (3D) space. We have developed a collagen matrix-based assay in a 96-well plate format that allows quantitative locomotion analyses of infected DCs in a 3D confinement over time. We report that active invasion of DCs by T. gondii tachyzoites induces enhanced migration of infected DCs in the collagen matrix. Parasites of genotype II induced superior DC migratory distances than type I parasites. Moreover, Toxoplasma-induced hypermigration of DCs was further potentiated in the presence of the CCR7 chemotactic cue CCL19. Blocking antibodies to integrins (CD11a, CD11b, CD18, CD29, CD49b) insignificantly affected migration of infected DCs in the 3D matrix, contrasting with their inhibitory effects on adhesion in 2D assays. Morphological analyses of infected DCs in the matrix were consistent with the acquisition of an amoeboid-like migratory phenotype. Altogether, the present data show that the Toxoplasma-induced hypermigratory phenotype in a 3D matrix is consistent with integrin-independent amoeboid DC migration with maintained responsiveness to chemotactic and chemokinetic cues. The data support the hypothesis that induction of amoeboid hypermigration and chemotaxis/chemokinesis in infected DCs potentiates the dissemination of T. gondii.
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- 2015
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13. Interleukin-6 secretion by astrocytes is dynamically regulated by PI3K-mTOR-calcium signaling.
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Simone Codeluppi, Teresa Fernandez-Zafra, Katalin Sandor, Jacob Kjell, Qingsong Liu, Mathew Abrams, Lars Olson, Nathanael S Gray, Camilla I Svensson, and Per Uhlén
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Medicine ,Science - Abstract
After contusion spinal cord injury (SCI), astrocytes become reactive and form a glial scar. While this reduces spreading of the damage by containing the area of injury, it inhibits regeneration. One strategy to improve the recovery after SCI is therefore to reduce the inhibitory effect of the scar, once the acute phase of the injury has passed. The pleiotropic cytokine interleukin-6 (IL-6) is secreted immediately after injury and regulates scar formation; however, little is known about the role of IL-6 in the sub-acute phases of SCI. Interestingly, IL-6 also promotes axon regeneration, and therefore its induction in reactive astrocytes may improve regeneration after SCI. We found that IL-6 is expressed by astrocytes and neurons one week post-injury and then declines. Using primary cultures of rat astrocytes we delineated the molecular mechanisms that regulate IL-6 expression and secretion. IL-6 expression requires activation of p38 and depends on NF-κB transcriptional activity. Activation of these pathways in astrocytes occurs when the PI3K-mTOR-AKT pathway is inhibited. Furthermore, we found that an increase in cytosolic calcium concentration was necessary for IL-6 secretion. To induce IL-6 secretion in astrocytes, we used torin2 and rapamycin to block the PI3K-mTOR pathway and increase cytosolic calcium, respectively. Treating injured animals with torin2 and rapamycin for two weeks, starting two weeks after injury when the scar has been formed, lead to a modest effect on mechanical hypersensitivity, limited to the period of treatment. These data, taken together, suggest that treatment with torin2 and rapamycin induces IL-6 secretion by astrocytes and may contribute to the reduction of mechanical hypersensitivity after SCI.
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- 2014
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14. RET PLCγ phosphotyrosine binding domain regulates Ca2+ signaling and neocortical neuronal migration.
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T Kalle Lundgren, Katsutoshi Nakahata, Nicolas Fritz, Paola Rebellato, Songbai Zhang, and Per Uhlén
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Medicine ,Science - Abstract
The receptor tyrosine kinase RET plays an essential role during embryogenesis in regulating cell proliferation, differentiation, and migration. Upon glial cell line-derived neurotrophic factor (GDNF) stimulation, RET can trigger multiple intracellular signaling pathways that in concert activate various downstream effectors. Here we report that the RET receptor induces calcium (Ca(2+)) signaling and regulates neocortical neuronal progenitor migration through the Phospholipase-C gamma (PLCγ) binding domain Tyr1015. This signaling cascade releases Ca(2+) from the endoplasmic reticulum through the inositol 1,4,5-trisphosphate receptor and stimulates phosphorylation of ERK1/2 and CaMKII. A point mutation at Tyr1015 on RET or small interfering RNA gene silencing of PLCγ block the GDNF-induced signaling cascade. Delivery of the RET mutation to neuronal progenitors in the embryonic ventricular zone using in utero electroporation reveal that Tyr1015 is necessary for GDNF-stimulated migration of neurons to the cortical plate. These findings demonstrate a novel RET mediated signaling pathway that elevates cytosolic Ca(2+) and modulates neuronal migration in the developing neocortex through the PLCγ binding domain Tyr1015.
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- 2012
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15. PfMDR1: mechanisms of transport modulation by functional polymorphisms.
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Pedro Eduardo Ferreira, Gabrielle Holmgren, Maria Isabel Veiga, Per Uhlén, Akira Kaneko, and José Pedro Gil
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Medicine ,Science - Abstract
ATP-Binding Cassette (ABC) transporters are efflux pumps frequently associated with multidrug resistance in many biological systems, including malaria. Antimalarial drug-resistance involves an ABC transporter, PfMDR1, a homologue of P-glycoprotein in humans. Twenty years of research have shown that several single nucleotide polymorphisms in pfmdr1 modulate in vivo and/or in vitro drug susceptibility. The underlying physiological mechanism of the effect of these mutations remains unclear. Here we develop structural models for PfMDR1 in different predicted conformations, enabling the study of transporter motion. Such analysis of functional polymorphisms allows determination of their potential role in transport and resistance. The bacterial MsbA ABC pump is a PfMDR1 homologue. MsbA crystals in different conformations were used to create PfMDR1 models with Modeller software. Sequences were aligned with ClustalW and analysed by Ali2D revealing a high level of secondary structure conservation. To validate a potential drug binding pocket we performed antimalarial docking simulations. Using aminoquinoline as probe drugs in PfMDR1 mutated parasites we evaluated the physiology underlying the mechanisms of resistance mediated by PfMDR1 polymorphisms. We focused on the analysis of well known functional polymorphisms in PfMDR1 amino acid residues 86, 184, 1034, 1042 and 1246. Our structural analysis suggested the existence of two different biophysical mechanisms of PfMDR1 drug resistance modulation. Polymorphisms in residues 86/184/1246 act by internal allosteric modulation and residues 1034 and 1042 interact directly in a drug pocket. Parasites containing mutated PfMDR1 variants had a significant altered aminoquinoline susceptibility that appears to be dependent on the aminoquinoline lipophobicity characteristics as well as vacuolar efflux by PfCRT. We previously described the in vivo selection of PfMDR1 polymorphisms under antimalarial drug pressure. Now, together with recent PfMDR1 functional reports, we contribute to the understanding of the specific structural role of these polymorphisms in parasite antimalarial drug response.
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- 2011
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16. Mapping genes for calcium signaling and their associated human genetic disorders.
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Matthias Hörtenhuber, Enrique M. Toledo, Erik Smedler, Ernest Arenas, Seth Malmersjö, Lauri Louhivuori, and Per Uhlén
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- 2017
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17. Table S1 from Membrane-Depolarizing Channel Blockers Induce Selective Glioma Cell Death by Impairing Nutrient Transport and Unfolded Protein/Amino Acid Responses
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Michael Andäng, Per Øyvind Enger, Sven Nelander, C. Theresa Vincent, Lukas Trantirek, Per Uhlén, Sten Linnarsson, A.F. Maarten Altelaar, Lene Uhrbom, Annika Jenmalm Jensen, Lars G.J. Hammarström, Anna-Lena Gustavsson, Martin Haraldsson, Anna Segerman, Voichita D. Marinescu, Jennifer M. Feenstra, Brittany B. Carson, Michaela Krafcikova, Gregorios Kyriatzis, Ivar Dehnisch, Nicolas Fritz, Linnéa Schmidt, Petra Sekyrova, Shimei Wee, Ercan Mutlu, Zuzana Sramkova, Gianluca Maddalo, and Mia Niklasson
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Small molecule screen data
- Published
- 2023
18. Supplementary Information from Membrane-Depolarizing Channel Blockers Induce Selective Glioma Cell Death by Impairing Nutrient Transport and Unfolded Protein/Amino Acid Responses
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Michael Andäng, Per Øyvind Enger, Sven Nelander, C. Theresa Vincent, Lukas Trantirek, Per Uhlén, Sten Linnarsson, A.F. Maarten Altelaar, Lene Uhrbom, Annika Jenmalm Jensen, Lars G.J. Hammarström, Anna-Lena Gustavsson, Martin Haraldsson, Anna Segerman, Voichita D. Marinescu, Jennifer M. Feenstra, Brittany B. Carson, Michaela Krafcikova, Gregorios Kyriatzis, Ivar Dehnisch, Nicolas Fritz, Linnéa Schmidt, Petra Sekyrova, Shimei Wee, Ercan Mutlu, Zuzana Sramkova, Gianluca Maddalo, and Mia Niklasson
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Supplementary Figure Legends and Materials and Methods
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- 2023
19. Figure S3 from Membrane-Depolarizing Channel Blockers Induce Selective Glioma Cell Death by Impairing Nutrient Transport and Unfolded Protein/Amino Acid Responses
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Michael Andäng, Per Øyvind Enger, Sven Nelander, C. Theresa Vincent, Lukas Trantirek, Per Uhlén, Sten Linnarsson, A.F. Maarten Altelaar, Lene Uhrbom, Annika Jenmalm Jensen, Lars G.J. Hammarström, Anna-Lena Gustavsson, Martin Haraldsson, Anna Segerman, Voichita D. Marinescu, Jennifer M. Feenstra, Brittany B. Carson, Michaela Krafcikova, Gregorios Kyriatzis, Ivar Dehnisch, Nicolas Fritz, Linnéa Schmidt, Petra Sekyrova, Shimei Wee, Ercan Mutlu, Zuzana Sramkova, Gianluca Maddalo, and Mia Niklasson
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Proteomics analysis
- Published
- 2023
20. Data from Membrane-Depolarizing Channel Blockers Induce Selective Glioma Cell Death by Impairing Nutrient Transport and Unfolded Protein/Amino Acid Responses
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Michael Andäng, Per Øyvind Enger, Sven Nelander, C. Theresa Vincent, Lukas Trantirek, Per Uhlén, Sten Linnarsson, A.F. Maarten Altelaar, Lene Uhrbom, Annika Jenmalm Jensen, Lars G.J. Hammarström, Anna-Lena Gustavsson, Martin Haraldsson, Anna Segerman, Voichita D. Marinescu, Jennifer M. Feenstra, Brittany B. Carson, Michaela Krafcikova, Gregorios Kyriatzis, Ivar Dehnisch, Nicolas Fritz, Linnéa Schmidt, Petra Sekyrova, Shimei Wee, Ercan Mutlu, Zuzana Sramkova, Gianluca Maddalo, and Mia Niklasson
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Glioma-initiating cells (GIC) are considered the underlying cause of recurrences of aggressive glioblastomas, replenishing the tumor population and undermining the efficacy of conventional chemotherapy. Here we report the discovery that inhibiting T-type voltage-gated Ca2+ and KCa channels can effectively induce selective cell death of GIC and increase host survival in an orthotopic mouse model of human glioma. At present, the precise cellular pathways affected by the drugs affecting these channels are unknown. However, using cell-based assays and integrated proteomics, phosphoproteomics, and transcriptomics analyses, we identified the downstream signaling events these drugs affect. Changes in plasma membrane depolarization and elevated intracellular Na+, which compromised Na+-dependent nutrient transport, were documented. Deficits in nutrient deficit acted in turn to trigger the unfolded protein response and the amino acid response, leading ultimately to nutrient starvation and GIC cell death. Our results suggest new therapeutic targets to attack aggressive gliomas. Cancer Res; 77(7); 1741–52. ©2017 AACR.
- Published
- 2023
21. Whole-Brain Three-Dimensional Imaging of RNAs at Single-Cell Resolution
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Shigeaki Kanatani, Judith C. Kreutzmann, Yue Li, Zoe West, Danai Vougesi Nikou, Jacob Lercke Skytte, Lea Lydolph Larsen, Daisuke H. Tanaka, Dagmara Kaczynska, Keishiro Fukumoto, Naofumi Uesaka, Tsutomu Tanabe, Ayako Miyakawa, Urmas Roostalu, Jacob Hecksher-Sørensen, and Per Uhlén
- Abstract
Whole-brain three-dimensional (3D) imaging is desirable to obtain a comprehensive and unbiased view of architecture and neural circuitry. However, current spatial analytic methods for brain RNAs are limited to thin sections or small samples. Here, we combined multiple new techniques to develop TRIC-DISCO, a new pipeline that allows imaging of RNA spatial distributions in whole adult mouse brains. First, we developed Tris-mediated retention ofin situhybridization signal during clearing (TRIC), which produces highly transparent tissue while maintaining the RNA signal intensities. We then combined TRIC with DISCO clearing (TRIC-DISCO) by controlling temperature during thein situhybridization chain reaction (isHCR) to ensure uniform whole-brain staining. This pipeline eliminates the requirements for both strict RNase-free environments and workflow-compatible RNase inhibitors. Our TRIC-DISCO pipeline enables simple and robust, single-cell, whole-brain, 3D imaging of transcriptional signatures, cell-identity markers, and noncoding RNAs across the entire brain.
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- 2022
22. Anatomy of the complete mouse eye vasculature in development and pathology explored by light-sheet fluorescence microscopy
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Luc Krimpenfort, Maria Garcia-Collado, Tom van Leeuwen, Filippo Locri, Anna-Liisa Luik, Antonio Queiro-Palou, Shigeaki Kanatani, Helder André, Per Uhlén, and Lars Jakobsson
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Eye development and function rely on precise establishment, regression and maintenance of its many sub-vasculatures. These crucial vascular properties have been extensively investigated in eye development and disease utilizing genetic and experimental mouse models. However, due to technical limitations, individual studies have often restricted their focus to one specific sub-vasculature. Here, we apply a workflow that allows for visualisation of complete vasculatures of mouse eyes of various developmental stages. Through tissue depigmentation, immunostaining, clearing and light-sheet fluorescence microscopy (LSFM) entire vasculatures of the retina, vitreous (hyaloids) and uvea were simultaneously imaged at high resolution. In silico dissection provided detailed information on their 3D architecture and interconnections. By this method we describe remodelling of the postnatal iris vasculature following its disconnection to the feeding hyaloid vasculature. In addition, we demonstrate examples of conventional and LSFM-mediated analysis of choroidal neovascularisation after laser-induced wounding, showing added value of the presented workflow in analysis of modelled eye disease. These advancements in visualisation and analysis of the respective eye vasculatures in development and complex eye disease open for novel observations of their functional interplay at a whole-organ level.
- Published
- 2022
23. Imaging cleared tissues made easy
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Shigeaki Kanatani and Per Uhlén
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Cell Biology ,Molecular Biology ,Biochemistry ,Biotechnology - Published
- 2022
24. Small-world connectivity dictates collective endothelial cell signaling
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Matthew D. Lee, Charlotte Buckley, Xun Zhang, Lauri Louhivuori, Per Uhlén, Calum Wilson, and John G. McCarron
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Multidisciplinary ,cardiovascular system ,Endothelial Cells ,Endothelium ,Signal Transduction - Abstract
Every blood vessel is lined by a single layer of highly specialized, yet adaptable and multifunctional endothelial cells. These cells, the endothelium, control vascular contractility, hemostasis, and inflammation and regulate the exchange of oxygen, nutrients, and waste products between circulating blood and tissue. To control each function, the endothelium processes endlessly arriving requests from multiple sources using separate clusters of cells specialized to detect specific stimuli. A well-developed but poorly understood communication system operates between cells to integrate multiple lines of information and coordinate endothelial responses. Here, the nature of the communication network has been addressed using single-cell Ca2+ imaging across thousands of endothelial cells in intact blood vessels. Cell activities were cross-correlated and compared to a stochastic model to determine network connections. Highly correlated Ca2+ activities occurred in scattered cell clusters, and network communication links between them exhibited unexpectedly short path lengths. The number of connections between cells (degree distribution) followed a power-law relationship revealing a scale-free network topology. The path length and degree distribution revealed an endothelial network with a “small-world” configuration. The small-world configuration confers particularly dynamic endothelial properties including high signal-propagation speed, stability, and a high degree of synchronizability. Local activation of small clusters of cells revealed that the short path lengths and rapid signal transmission were achieved by shortcuts via connecting extensions to nonlocal cells. These findings reveal that the endothelial network design is effective for local and global efficiency in the interaction of the cells and rapid and robust communication between endothelial cells in order to efficiently control cardiovascular activity.
- Published
- 2022
25. Glyphosate-based herbicide induces long-lasting impairment in neuronal and glial differentiation
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Luã Reis, Marilena Raciti, Patricia González Rodriguez, Bertrand Joseph, Ibrahim Al Rayyes, Per Uhlén, Anna Falk, Suzana Telles da Cunha Lima, and Sandra Ceccatelli
- Subjects
Neurons ,Herbicides ,Health, Toxicology and Mutagenesis ,Neurogenesis ,Glycine ,Humans ,General Medicine ,Management, Monitoring, Policy and Law ,Toxicology - Abstract
Glyphosate-based herbicides (GBH) are among the most sold pesticides in the world. There are several formulations based on the active ingredient glyphosate (GLY) used along with other chemicals to improve the absorption and penetration in plants. The final composition of commercial GBH may modify GLY toxicological profile, potentially enhancing its neurotoxic properties. The developing nervous system is particularly susceptible to insults occurring during the early phases of development, and exposure to chemicals in this period may lead to persistent impairments on neurogenesis and differentiation. The aim of this study was to evaluate the long-lasting effects of a sub-cytotoxic concentration, 2.5 parts per million of GBH and GLY, on the differentiation of human neuroepithelial stem cells (NES) derived from induced pluripotent stem cells (iPSC). We treated NES cells with each compound and evaluated the effects on key cellular processes, such as proliferation and differentiation in daughter cells never directly exposed to the toxicants. We found that GBH induced a more immature neuronal profile associated to increased PAX6, NESTIN and DCX expression, and a shift in the differentiation process toward glial cell fate at the expense of mature neurons, as shown by an increase in the glial markers GFAP, GLT1, GLAST and a decrease in MAP2. Such alterations were associated to dysregulation of key genes critically involved in neurogenesis, including PAX6, HES1, HES5, and DDK1. Altogether, the data indicate that subtoxic concentrations of GBH, but not of GLY, induce long-lasting impairments on the differentiation potential of NES cells.
- Published
- 2022
26. Disrupted Cacna1c gene expression perturbs spontaneous Ca 2+ activity causing abnormal brain development and increased anxiety
- Author
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Erik Smedler, Lauri Louhivuori, Roman A. Romanov, Débora Masini, Ivar Dehnisch Ellström, Chungliang Wang, Martino Caramia, Zoe West, Songbai Zhang, Paola Rebellato, Seth Malmersjö, Irene Brusini, Shigeaki Kanatani, Gilberto Fisone, Tibor Harkany, and Per Uhlén
- Subjects
Multidisciplinary - Abstract
Significance The gene CACNA1C encodes for a calcium channel that has been linked to various psychiatric conditions, including schizophrenia and bipolar disorder, through hitherto unknown cellular mechanisms. Here, we report that deletion of Cacna1c in neurons of the developing brain disrupts spontaneous calcium activity and causes abnormal brain development and anxiety. Our results indicate that marginally alterations in the expression level of Cacna1c have major effects on the intrinsic spontaneous calcium activity of neural progenitors that play a crucial role in brain development. Thus, Cacna1c acts as a molecular switch that can increase susceptibility to psychiatric disease.
- Published
- 2022
27. Scalable in situ single-cell profiling by electrophoretic capture of mRNA using EEL FISH
- Author
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Lars E. Borm, Alejandro Mossi Albiach, Camiel C. A. Mannens, Jokubas Janusauskas, Ceren Özgün, David Fernández-García, Rebecca Hodge, Francisca Castillo, Charlotte R. H. Hedin, Eduardo J. Villablanca, Per Uhlén, Ed S. Lein, Simone Codeluppi, and Sten Linnarsson
- Subjects
Biomedical Engineering ,Molecular Medicine ,Bioengineering ,Applied Microbiology and Biotechnology ,Biotechnology - Abstract
Methods to spatially profile the transcriptome are dominated by a trade-off between resolution and throughput. Here we develop a method named Enhanced ELectric Fluorescence in situ Hybridization (EEL FISH) that can rapidly process large tissue samples without compromising spatial resolution. By electrophoretically transferring RNA from a tissue section onto a capture surface, EEL speeds up data acquisition by reducing the amount of imaging needed, while ensuring that RNA molecules move straight down toward the surface, preserving single-cell resolution. We apply EEL on eight entire sagittal sections of the mouse brain and measure the expression patterns of up to 440 genes to reveal complex tissue organization. Moreover, EEL can be used to study challenging human samples by removing autofluorescent lipofuscin, enabling the spatial transcriptome of the human visual cortex to be visualized. We provide full hardware specifications, all protocols and complete software for instrument control, image processing, data analysis and visualization.
- Published
- 2022
28. Mechanisms underlying the generation of autonomorespiratory coupling amongst the respiratory central pattern generator, sympathetic oscillators, and cardiovagal premotoneurons
- Author
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M. Gazi Yasargil, George Zaki Ghali, Adriana Lima, Stefanos Voglis, Emma Glover, Jennifer Humphrey, Per Uhlén, Marton König, Michael George Zaki Ghali, Michael W. McDermott, Robert F. Spetzler, and Henry Brem
- Subjects
Sympathetic Nervous System ,Efferent ,Pre-Bötzinger complex ,01 natural sciences ,Efferent nerve ,lcsh:RC321-571 ,Midbrain ,Cardiovascular Physiological Phenomena ,03 medical and health sciences ,0302 clinical medicine ,Neural Pathways ,Animals ,Humans ,lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry ,Neurons ,Central chemoreceptors ,Chemistry ,General Neuroscience ,Respiration ,Central pattern generator ,Brain ,General Medicine ,Rostral ventrolateral medulla ,Respiratory Center ,030205 complementary & alternative medicine ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,nervous system ,Central Pattern Generators ,mechanisms|genesis|sympathorespiratory|coupling|modulation|cardiovagal|rostral ventrolateral medulla|sympathetic|hypercapnia|hypoxia ,Neuroscience ,Metencephalon - Abstract
The respiratory rhythm and pattern and sympathetic and parasympathetic outflows are generated by distinct, though overlapping, propriobulbar arrays of neuronal microcircuit oscillators constituting networks utilizing mutual excitatory and inhibitory neuronal interactions, residing principally within the metencephalon and myelencephalon, and modulated by synaptic influences from the cerebrum, thalamus, hypothalamus, cerebellum, and mesencephalon and ascending influences deriving from peripheral stimuli relayed by cranial nerve afferent axons. Though the respiratory and cardiovascular regulatory effector mechanisms utilize distinct generators, there exists significant overlap and interconnectivity amongst and between these oscillators and pathways, evidenced reciprocally by breathing modulation of sympathetic oscillations and sympathetic modulation of neural breathing. These coupling mechanisms are well-demonstrated coordinately in sympathetic- and respiratory-related central neuronal and efferent neurogram recordings and quantified by the findings of cross-correlation, spectra, and coherence analyses, combined with empirical interventions including lesioning and pharmacological agonist and antagonist microinjection studies, baroloading, barounloading, and hypoxic and/or hypercapnic peripheral and/or central chemoreceptor stimulation. Sympathetic and parasympathetic central neuronal and efferent neural discharge recordings evidence classic fast rhythms produced by propriobulbar neuronal networks located within the medullary division of the lateral tegmental field, coherent with cardiac sympathetic nerve discharge. These neural efferent nerve discharges coordinately evidence slow synchronous oscillations, constituted by Traube Hering (i.e., high frequency), Mayer wave (i.e., medium or low frequency), and vasogenic autorhythmicity (i.e., very low frequency) wave spectral bands. These oscillations contribute to coupling neural breathing, sympathetic oscillations, and parasympathetic cardiovagal premotoneuronal activity. The mechanisms underlying the origins of and coupling amongst, these waves remains to be unresolved.
- Published
- 2020
29. Neurotransmitters and Endothelins Acting on Radial Glial G-Protein-Coupled Receptors Are, Through Proteolytic NRG/ErbB4 Activation, Able to Modify the Migratory Behavior of Neocortical Cells and Mediate Bipolar-to-Multipolar Transition
- Author
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Tommy Nordström, Lauri Louhivuori, Per Uhlén, Karl E.O. Åkerman, Pauli M. Turunen, Verna Louhivuori, and Ibrahim Al Rayyes
- Subjects
medicine.hormone ,Receptor, ErbB-4 ,Receptor, Metabotropic Glutamate 5 ,Neocortex ,Cell Communication ,Biology ,Receptors, G-Protein-Coupled ,Endothelins ,03 medical and health sciences ,0302 clinical medicine ,TRPC3 ,Neural Stem Cells ,Cell Movement ,Muscarinic acetylcholine receptor ,medicine ,Animals ,Receptor ,Cell Shape ,TRPC ,Neuregulins ,TRPC Cation Channels ,030304 developmental biology ,G protein-coupled receptor ,Neurons ,Neurotransmitter Agents ,0303 health sciences ,Metabotropic glutamate receptor 5 ,Cell Biology ,Hematology ,Cell biology ,Mice, Inbred C57BL ,nervous system ,Proteolysis ,Neuregulin ,Neuroglia ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Cell-cell communication plays a central role in the guidance of migrating neurons during the development of the cerebral cortex. Neuregulins (NRGs) are essential mediators for migration and maintenance of the radial glial scaffold. We show, in this study that soluble NRG reduces neuronal motility, causes transition of bipolar cells to multipolar ones, and induces neuronal mitosis. Blocking the NRG receptor, ErbB4, results in reduction of neuron-neuron and neuron-radial glial contacts and causes an increase in neuronal motility. Blocking the radial glial metabotropic glutamate receptor 5 (mGluR5), the nonselective cation channel transient receptor potential 3 (TRPC3), or matrix metalloproteinases (MMPs) results in similar effects as ErbB4 blockade. Soluble NRG counteract the changes in motility pattern. Stimulation of other radial glial G-protein-coupled receptors (GPCRs), such as muscarinic acetylcholine receptors or endothelin receptors counteract all the effect of mGluR5 blockade, but not that of ErbB4, TRPC3, and MMP blockade. The results indicate that neurotransmitters and endothelins acting on radial glial GPCRs are, through proteolytic NRG/ErbB4 activation, able to modify the migratory behavior of neurons.
- Published
- 2020
30. STEM-19. LIVE DETECTION OF NEURAL STEM AND GLIOBLASTOMA CELLS BY A LUMINESCENT CONJUGATED OLIGOTHIOPHENE DERIVATIVE
- Author
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Shirin Ilkhanizadeh, Aileen Gracias, Andreas K O Åslund, Marcus Bäck, Rozalyn Simon, Christina Neofytou, Vilma Rraklli, Bianca Migliori, Edel Kavanagh, Sven Nelander, Bengt Westermark, Lene Uhrbom, Karin Forsberg-Nilsson, Ana I Teixeira, Peter Konradsson, Per Uhlén, Johan Holmberg, Bertrand Joseph, K Peter R Nilsson, and Ola Hermanson
- Subjects
Cancer Research ,Oncology ,Neurology (clinical) - Abstract
Glioblastoma (GBM) is an aggressive nervous system tumor with a mean survival time of 12-14 months. Cells with neural stem cell-like properties can be derived from GBM tumors. These cells seem to escape conventional irradiation treatment, chemotherapy, and surgery, and may play a crucial role for relapse. It is therefore urgent to develop novel approaches for reliable detection of neural stem cell-like cells in GBM. Here we report a luminescent conjugated oligothiophene (LCO), named GlioStem (p-HTMI), for non-invasive and non-amplified real-time detection of live human patient-derived GBM cells and embryonic neural stem/progenitor cells (NSPCs). Within a maximum of 10 minutes after administration of the molecule in vitro, in the existing media, fluorescence emission was observed without any modulation of the cells or additional vehicle, resulting in efficient detection of cytoplasmic luminescent signal in NSPCs or GBM cells from rodents and humans, detectable at Alexa488/GFP wavelength. GlioStem is functionalized with a methylated imidazole moiety resembling the side chain of histidine/histamine, and non-methylated analogues were not functional. In vitro, GlioStem was shown to identify fetal cortical NSPCs from rat (FGF2-expanded), embryonic stem cell-derived NSPCs from mouse (FGF2/EGF-expanded), and FGF2-exposed C6 glioma cell cultures from rat, but not any other cell types investigated. Cell sorting experiments of patient-derived, FGF2/EGF-expanded GBM cells demonstrated that GlioStem in addition to NSPC-markers like Nestin and Sox2 labeled the same population (overlap > 90%) of cells as CD271, a proposed marker for stem cell-like cells and rapidly migrating cells in glioblastoma. Our results suggest that the LCO GlioStem is a versatile tool for immediate and selective detection of subpopulations of neural stem and glioma cells.
- Published
- 2022
31. Arg1+ microglia are critical for shaping cognition in female mice
- Author
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Patricia González-Rodríguez, Michael T. Heneka, Lily Keane, Adriana-Natalia Murgoci, José L. Venero, Irene García-Domínguez, Guillermo Vázquez-Cabrera, Marie-Ève Tremblay, Eva M. Pérez-Villegas, Irene Martinez-Gallego, Mikko Airavaara, Dario Tejera, Rocío Ruiz, Isabel María Alonso-Bellido, Bertrand Joseph, Shigeaki Kanatani, Per Uhlén, Ahmed M. Osman, Kathleen Grabert, Javier Avila-Cariño, Antonio Rodríguez-Moreno, Vassilis Stratoulias, Klas Blomgren, David Brodin, José A. Armengol, Mathilde Cheray, and Nathalie Vernoux
- Subjects
Apolipoprotein E ,0303 health sciences ,Dendritic spine ,Microglia ,Hippocampus ,Long-term potentiation ,Biology ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,nervous system ,Forebrain ,medicine ,Cholinergic ,Cholinergic neuron ,Neuroscience ,030217 neurology & neurosurgery ,030304 developmental biology - Abstract
Diversity within microglia, the resident brain immune cells, is reported. Whether microglial subsets constitute different subtypes with intrinsic properties and unique functions has not been fully elucidated. Here, we describe a microglial subtype characterized by the expression of the enzyme Arginase-1, i.e. Arg1+microglia, which is found predominantly in the cholinergic neuron-rich forebrain region during early postnatal development. Arg1+ microglia are frequently observed in close apposition to neurons and exhibit a distinctive molecular signature reflecting a reactive profile. Arg1 deficiency in microglia results in impaired dendritic spine maturation in the hippocampus where cholinergic neurons project, and cognitive behavioural deficiencies in female mice. Our results expand on microglia diversity and provide insights into distinctive spatiotemporal functions exerted by microglial subtypes.
- Published
- 2021
32. Retraction: Ghali MGZ
- Author
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Michael G Z, Ghali, George Zaki, Ghali, Adriana, Lima, Michael, McDermott, Emma, Glover, Stefanos, Voglis, Jennifer, Humphrey, Marton Skog Steinberger, König, Henry, Brem, Per, Uhlén, Robert F, Spetzler, and M Gazi, Yasargil
- Abstract
No abstract present.
- Published
- 2021
33. The T-type Ca2+ Channel Cav3.2 Regulates Differentiation of Neural Progenitor Cells during Cortical Development via Caspase-3
- Author
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Songbai Zhang, Ernest Arenas, Shigeaki Kanatani, Lauri Louhivuori, Dagmara Kaczynska, Ibrahim Al Rayyes, Ola Hermanson, Carlos Villaescusa, Paola Rebellato, Anna Falk, and Per Uhlén
- Subjects
0301 basic medicine ,biology ,Chemistry ,General Neuroscience ,Cellular differentiation ,Neurogenesis ,T-type calcium channel ,Caspase 3 ,Neural stem cell ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Downregulation and upregulation ,Knockout mouse ,CACNA1H ,biology.protein ,030217 neurology & neurosurgery - Abstract
Here we report that the low-voltage-dependent T-type calcium (Ca2+) channel Cav3.2, encoded by the CACNA1H gene, regulates neuronal differentiation during early embryonic brain development through activating caspase-3. At the onset of neuronal differentiation, neural progenitor cells exhibited spontaneous Ca2+ activity. This activity strongly correlated with the upregulation of CACNA1H mRNA. Cells exhibiting robust spontaneous Ca2+ signaling had increased caspase-3 activity unrelated to apoptosis. Inhibition of Cav3.2 by drugs or viral CACNA1H knock down resulted in decreased caspase-3 activity followed by suppressed neurogenesis. In contrast, when CACNA1H was overexpressed, increased neurogenesis was detected. Cortical slices from Cacna1h knockout mice showed decreased spontaneous Ca2+ activity, a significantly lower protein level of cleaved caspase-3, and microanatomical abnormalities in the subventricular/ventricular and cortical plate zones when compared to their respective embryonic controls. In summary, we demonstrate a novel relationship between Cav3.2 and caspase-3 signaling that affects neurogenesis in the developing brain.
- Published
- 2019
34. BCG‐induced cytokine release in bladder cancer cells is regulated by Ca 2+ signaling
- Author
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Per Uhlén, T. Kalle Lundgren, Sara M. Mangsbo, Rahim Kaba, Abolfazl Hosseini, Simone Codeluppi, Arad Hosseini, Marie Karlsson, Mototsugu Oya, Manuel Varas-Godoy, Peter Wiklund, Lauri Louhivuori, Ayako Miyakawa, Nobuyuki Tanaka, Navid Soltani, Cristian Ibarra, and Songbai Zhang
- Subjects
0301 basic medicine ,Cancer Research ,medicine.medical_treatment ,calcium signaling ,lcsh:RC254-282 ,Proinflammatory cytokine ,03 medical and health sciences ,Cytosol ,0302 clinical medicine ,Cell Line, Tumor ,Urologi och njurmedicin ,Genetics ,medicine ,Humans ,Urology and Nephrology ,BCG ,TLR4 ,Interleukin 8 ,Research Articles ,Calcium signaling ,urinary bladder cancer ,Bladder cancer ,business.industry ,Interleukin-8 ,NF-kappa B ,Cancer ,General Medicine ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,medicine.disease ,Mycobacterium bovis ,Toll-Like Receptor 4 ,030104 developmental biology ,Cytokine ,Urinary Bladder Neoplasms ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,Cytokines ,Molecular Medicine ,Calcium ,Signal transduction ,business ,Research Article - Abstract
Bacillus Calmette-Guerin (BCG) is widely used in the clinic to effectively treat superficial urinary bladder cancer. However, a significant proportion of patients who fail to respond to BCG risk cystectomy or death. Though more than 3 million cancer treatments with BCG occur annually, surprisingly little is known about the initial signaling cascades activated by BCG. Here, we report that BCG induces a rapid intracellular Ca2+ (calcium ion) signal in bladder cancer cells that is essential for activating the transcription factor nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappa B) and for synthesizing and secreting proinflammatory cytokines, including interleukin 8 (IL-8). A similar Ca2+ response was observed when cells were exposed to the supernatant of BCG. Studying cellular molecular mechanisms involved in the BCG signaling event, we found pivotal roles for phospholipase C and the Toll-like receptor 4. Further assessment revealed that this signaling pathway induces synthesis of IL-8, whereas exocytosis appeared to be controlled by global Ca2+ signaling. These results shed new light on the molecular mechanisms underlying BCG treatment of bladder cancer, which can help in improving therapeutic efficacy and reducing adverse side effects.
- Published
- 2018
35. Single-cell RNA-seq analysis reveals the platinum resistance gene COX7B and the surrogate marker CD63
- Author
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Aparna Reddy, Nobuyuki Tanaka, Takeo Kosaka, Koichiro Ogihara, Kaneyasu Nishimura, Juha Kere, Hiroshi Hongo, Ernest Arenas, Ayako Miyakawa, Shintaro Katayama, Naoya Niwa, Ryuichi Mizuno, Mototsugu Oya, Per Uhlén, Eiji Kikuchi, and Shuji Mikami
- Subjects
0301 basic medicine ,Cancer Research ,medicine.medical_treatment ,Antineoplastic Agents ,Drug resistance ,Biology ,Electron Transport Complex IV ,Transcriptome ,03 medical and health sciences ,Cell Line, Tumor ,CD63 ,tumor heterogeneity ,medicine ,Humans ,Radiology, Nuclear Medicine and imaging ,single‐cell RNA‐seq ,Original Research ,Cancer Biology ,Cisplatin ,Gene knockdown ,Chemotherapy ,Sequence Analysis, RNA ,Tetraspanin 30 ,Cancer ,medicine.disease ,COX7B ,platinum resistance ,female genital diseases and pregnancy complications ,3. Good health ,030104 developmental biology ,Urinary Bladder Neoplasms ,Oncology ,Drug Resistance, Neoplasm ,Cancer cell ,Cancer research ,Single-Cell Analysis ,medicine.drug - Abstract
Cancers acquire resistance to systemic treatment with platinum‐based chemotherapy (eg, cisplatin [CDDP]) as a result of a dynamic intratumoral heterogeneity (ITH) and clonal repopulation. However, little is known about the influence of chemotherapy on ITH at the single‐cell level. Here, mapping the transcriptome of cancers treated with CDDP by scRNA‐seq, we uncovered a novel gene, COX7B, associated with platinum‐resistance, and surrogate marker, CD63. Knockdown of COX7B in cancer cells decreased the sensitivity of CDDP whereas overexpression recovered the sensitivity of CDDP. Low COX7B levels correlated with higher mortality rates in patients with various types of cancer and were significantly associated with poor response to chemotherapy in urinary bladder cancer. Tumor samples from patients, who underwent CDDP therapy, showed decreased COX7B protein levels after the treatment. Analyzing scRNA‐seq data from platinum‐naïve cancer cells demonstrated a low‐COX7B subclone that could be sorted out from bulk cancer cells by assaying CD63. This low‐COX7B subclone behaved as cells with acquired platinum‐resistance when challenged to CDDP. Our results offer a new transcriptome landscape of platinum‐resistance that provides valuable insights into chemosensitivity and drug resistance in cancers, and we identify a novel platinum resistance gene, COX7B, and a surrogate marker, CD63.
- Published
- 2018
36. Correction to: Human ex vivo spinal cord slice culture as a useful model of neural development, lesion, and allogeneic neural cell therapy
- Author
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Elisabet Åkesson, Chenhong Lin, Åsa Ekblad-Nordberg, Lena Holmberg, Jia Liu, Erika Vázquez-Juárez, Per Uhlén, Teresa Fernandez-Zafra, Xiaofei Li, Maria Lindskog, Cinzia Calzarossa, and Simone Codeluppi
- Subjects
Pathology ,medicine.medical_specialty ,Cell- and Tissue-Based Therapy ,Medicine (miscellaneous) ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,lcsh:Biochemistry ,Lesion ,medicine ,Animals ,Humans ,lcsh:QD415-436 ,Neural cell ,Spinal cord slice ,Spinal Cord Injuries ,Neurons ,lcsh:R5-920 ,business.industry ,Hematopoietic Stem Cell Transplantation ,Correction ,Cell Biology ,Spinal Cord ,Molecular Medicine ,medicine.symptom ,Stem cell ,lcsh:Medicine (General) ,business ,Neural development ,Ex vivo - Abstract
There are multiple promising treatment strategies for central nervous system trauma and disease. However, to develop clinically potent and safe treatments, models of human-specific conditions are needed to complement in vitro and in vivo animal model-based studies.We established human brain stem and spinal cord (cross- and longitudinal sections) organotypic cultures (hOCs) from first trimester tissues after informed consent by donor and ethical approval by the Regional Human Ethics Committee, Stockholm (lately referred to as Swedish Ethical Review Authority), and The National Board of Health and Welfare, Sweden. We evaluated the stability of hOCs with a semi-quantitative hOC score, immunohistochemistry, flow cytometry, CaThe spinal cord hOCs presented relatively stable features during 7-21 days in vitro (DIV) (except a slightly increased cell proliferation and activated glial response). After contusion injury performed at 7 DIV, a significant reduction of the hOC score, increase of the activated caspase-3We conclude that human spinal cord slice cultures have potential for future structural and functional studies of human spinal cord development, injury, and treatment strategies.
- Published
- 2020
37. Human ex vivo spinal cord slice culture as a useful model of neural development, lesion, and allogeneic neural cell therapy
- Author
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Teresa Fernandez-Zafra, Åsa Ekblad-Nordberg, Elisabet Åkesson, Xiaofei Li, Simone Codeluppi, Erika Vázquez-Juárez, Jia Liu, Lena Holmberg, Per Uhlén, Maria Lindskog, Cinzia Calzarossa, and Chenhong Lin
- Subjects
Pathology ,medicine.medical_specialty ,medicine.medical_treatment ,Central nervous system ,Population ,Medicine (miscellaneous) ,Spinal cord injury ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,lcsh:Biochemistry ,Human organotypic culture ,medicine ,lcsh:QD415-436 ,education ,education.field_of_study ,lcsh:R5-920 ,Stem cell therapy ,business.industry ,Research ,Cell Biology ,Stem-cell therapy ,medicine.disease ,Spinal cord ,medicine.anatomical_structure ,Molecular Medicine ,Stem cell ,business ,lcsh:Medicine (General) ,Neural development ,Ex vivo - Abstract
BackgroundThere are multiple promising treatment strategies for central nervous system trauma and disease. However, to develop clinically potent and safe treatments, models of human-specific conditions are needed to complement in vitro and in vivo animal model-based studies.MethodsWe established human brain stem and spinal cord (cross- and longitudinal sections) organotypic cultures (hOCs) from first trimester tissues after informed consent by donor and ethical approval by the Regional Human Ethics Committee, Stockholm (lately referred to as Swedish Ethical Review Authority), and The National Board of Health and Welfare, Sweden. We evaluated the stability of hOCs with a semi-quantitative hOC score, immunohistochemistry, flow cytometry, Ca2+signaling, and electrophysiological analysis. We also applied experimental allogeneic human neural cell therapy after injury in the ex vivo spinal cord slices.ResultsThe spinal cord hOCs presented relatively stable features during 7–21 days in vitro (DIV) (except a slightly increased cell proliferation and activated glial response). After contusion injury performed at 7 DIV, a significant reduction of the hOC score, increase of the activated caspase-3+cell population, and activated microglial populations at 14 days postinjury compared to sham controls were observed. Such elevation in the activated caspase-3+population and activated microglial population was not observed after allogeneic human neural cell therapy.ConclusionsWe conclude that human spinal cord slice cultures have potential for future structural and functional studies of human spinal cord development, injury, and treatment strategies.
- Published
- 2020
38. Mending Fences: Na,K-ATPase signaling via Ca
- Author
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Anita, Aperia, Hjalmar, Brismar, and Per, Uhlén
- Abstract
Na,K-ATPase is a ubiquitous multifunctional protein that acts both as an ion pump and as a signal transducer. The signaling function is activated by ouabain in non-toxic concentrations. In epithelial cells the ouabain-bound Na,K-ATPase connects with the inositol 1,4,5-trisphosphate receptor via a short linear motif to activate low frequency Ca2+ oscillations. Within a couple of minutes this ouabain mediated signal has resulted in phosphorylation or dephosphorylation of 2580 phospho-sites. Proteins that control cell proliferation and cell adhesion and calmodulin regulated proteins are enriched among the ouabain phosphor-regulated proteins. The inositol 1,4,5-trisphosphate receptor and the stromal interaction molecule, which are both essential for the initiation of Ca2+ oscillations, belong to the ouabain phosphor-regulated proteins. Downstream effects of the ouabain-evoked Ca2+ signal in epithelial cells include interference with the intrinsic mitochondrial apoptotic process and stimulation of embryonic growth processes. The dual function of Na,K-ATPase as an ion pump and a signal transducer is now well established and evaluation of the physiological and pathophysiological consequences of this universal signal emerges as an urgent topic for future studies.
- Published
- 2020
39. Decision letter: Aberrant calcium channel splicing drives defects in cortical differentiation in Timothy syndrome
- Author
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Per Uhlén and Anita Bhattacharyya
- Subjects
Calcium channel ,RNA splicing ,Timothy syndrome ,medicine ,Biology ,medicine.disease ,Cell biology - Published
- 2019
40. Improved Pathological Examination of Tumors with 3D Light-Sheet Microscopy
- Author
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Per Uhlén and Nobuyuki Tanaka
- Subjects
0301 basic medicine ,Microscopy ,Cancer Research ,Pathology ,medicine.medical_specialty ,business.industry ,food and beverages ,Cancer ,medicine.disease ,03 medical and health sciences ,030104 developmental biology ,Oncology ,Neoplasms ,Light sheet fluorescence microscopy ,medicine ,Humans ,business ,Pathological - Abstract
Light-sheet microscopy offers new possibilities to efficiently visualize large tissue samples in three dimensions. Volumetric 3D imaging can uncover detailed information about the inner landscape of tumors, which can improve cancer diagnosis and therapy. This Forum article highlights the advantages of using light-sheet microscopy for pathological examinations of intact tumor specimens.
- Published
- 2018
41. Light-sheet microscopy: Diagnosing intratumoral heterogeneity of intact tumors in three-dimension
- Author
-
Dagmara Kaczynska, Ayako Miyakawa, Shigeaki Kanatani, Lauri Louhivuori, Mototsugu Oya, Per Uhlén, Nobuyuki Tanaka, and Peter Wiklund
- Subjects
Dimension (vector space) ,business.industry ,Urology ,Light sheet fluorescence microscopy ,Medicine ,business ,Biomedical engineering - Published
- 2018
42. Whole-tissue biopsy phenotyping of three-dimensional tumours reveals patterns of cancer heterogeneity
- Author
-
Dagmara Kaczynska, Karl Deisseroth, Carlos Fernández Moro, Przemysław Mitura, Loránd L. Kis, Sara Corvigno, Ayako Miyakawa, Kazuhiro Matsumoto, Shigeaki Kanatani, Pauliina Kronqvist, Carina Strell, Lauri Louhivuori, Mototsugu Oya, Nobuyuki Tanaka, Arne Östman, Claes Lindh, Joseph W. Carlson, Patrick Micke, Andrzej Stepulak, Cecilia Sahlgren, Raju Tomer, Per Uhlén, Peter Wiklund, Artur Mezheyeuski, Hanna Dahlstrand, Johan Hartman, and Soft Tissue Biomech. & Tissue Eng.
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,medicine.diagnostic_test ,Tumour heterogeneity ,Genetic heterogeneity ,Angiogenesis ,Biomedical Engineering ,Medicine (miscellaneous) ,Cancer ,Bioengineering ,Biology ,SDG 3 – Goede gezondheid en welzijn ,medicine.disease ,Computer Science Applications ,03 medical and health sciences ,030104 developmental biology ,SDG 3 - Good Health and Well-being ,Biopsy ,Cancer cell ,medicine ,Epigenetics ,Biotechnology ,Tissue biopsy - Abstract
Intratumoral heterogeneity is a critical factor when diagnosing and treating patients with cancer. Marked differences in the genetic and epigenetic backgrounds of cancer cells have been revealed by advances in genome sequencing, yet little is known about the phenotypic landscape and the spatial distribution of intratumoral heterogeneity within solid tumours. Here, we show that three-dimensional light-sheet microscopy of cleared solid tumours can identify unique patterns of phenotypic heterogeneity, in the epithelial-to-mesenchymal transition and in angiogenesis, at single-cell resolution in whole formalin-fixed paraffin-embedded (FFPE) biopsy samples. We also show that cleared FFPE samples can be re-embedded in paraffin after examination for future use, and that our tumour-phenotyping pipeline can determine tumour stage and stratify patient prognosis from clinical samples with higher accuracy than current diagnostic methods, thus facilitating the design of more efficient cancer therapies. A method that identifies patterns of tumour heterogeneity in intact biopsy samples using 3D light-sheet microscopy stratifies patients by tumour stage.
- Published
- 2017
43. Predicting a tumour's drug uptake
- Author
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Shigeaki Kanatani, Lauri Louhivuori, and Per Uhlén
- Subjects
0301 basic medicine ,Volumetric imaging ,Computational model ,Tumour heterogeneity ,Chemistry ,Biomedical Engineering ,Medicine (miscellaneous) ,Bioengineering ,Blood flow ,Drug uptake ,Computer Science Applications ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Three dimensional imaging ,030220 oncology & carcinogenesis ,Neoplasms ,Hydrodynamics ,Humans ,Biotechnology ,Biomedical engineering - Abstract
Computational modelling, informed by data from 3D volumetric imaging of transparent and intact whole-tumour samples, predicts blood flow and the spatial distribution of drug uptake in tumours.
- Published
- 2019
44. Recombinant spider silk protein matrices facilitate multi-analysis of calcium-signaling in neural stem cell-derived AMPA-responsive neurons
- Author
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Paola Rebellato, Michalina Lewicka, Per Uhlén, Jakub Lewicki, Anna Rising, and Ola Hermanson
- Subjects
nervous system ,Tissue engineering ,Chemistry ,Growth factor ,medicine.medical_treatment ,medicine ,Context (language use) ,Spider silk ,Stem cell ,Progenitor cell ,Neural stem cell ,WNT3A ,Cell biology - Abstract
Neural progenitors or stem cells (NSCs) show great promise in drug discovery and clinical application. Yet few efforts have been made to optimize biocompatible materials for such cells to be expanded and used in clinical conditions. We have previously demonstrated that NSCs are readily cultured on substrates of certain recombinant spider silk protein without addition of animal- or human-derived components. The question remains however whether this material allows differentiation into functional neurons and glia, and whether such differentiation can take place also when the NSCs are cultured within the material in a pseudo-3D context. Here we demonstrate that “foam”-like structures generated from recombinant spider silk protein (4RepCT) provided excellent matrices for the generation and multicellular analysis of functional excitatory neurons from NSCs without addition of animal- or human-derived components. NSCs isolated from the cerebral cortices of rat embryos were cultured on either 4RepCT matrices shaped as foam-like structures without coating, or on conventional polystyrene plates coated with poly-L-ornithine and fibronectin. Upon treatment with recombinant proteins including the growth factor BMP4 or a combination of BMP4 and the signaling factor Wnt3a, the cortical NSCs cultured in 4RepCT foam-like structures differentiated efficiently into neurons that responded to glutamate receptor agonists, such as AMPA, to at least the same extent as control cultures. Matrices derived from recombinant spider silk proteins thus provide a functional microenvironment for neural stem cells without any animal- or human-derived components, and can be employed in the development of new strategies in stem cell research and tissue engineering.
- Published
- 2019
45. Retraction: Ghali MGZ, et al. Mechanisms underlying the generation of autonomorespiratory coupling amongst the respiratory central pattern generator, sympathetic oscillators, and cardiovagal premotoneurons. Journal of Integrative Neuroscience. 2020; 19: 521–560
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Henry Brem, Stefanos Voglis, Marton König, Jennifer Humphrey, Michael W. McDermott, Per Uhlén, George Zaki Ghali, Adriana Lima, M. Gazi Yasargil, Emma Glover, Michael George Zaki Ghali, and Robert F. Spetzler
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Physics ,Coupling (electronics) ,Integrative neuroscience ,General Neuroscience ,Central pattern generator ,Neurosciences. Biological psychiatry. Neuropsychiatry ,General Medicine ,Neuroscience ,RC321-571 - Abstract
No abstract present.
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- 2021
46. Notch activation in the mouse mammary luminal lineage leads to ductal hyperplasia and altered partitioning of luminal cell subtypes
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Per Uhlén, Indira V. Chivukula, Yee Peng Phoon, Shigeaki Kanatani, Raoul Kuiper, Urban Lendahl, and Yat Long Tsoi
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0301 basic medicine ,Notch signaling pathway ,Mammary Neoplasms, Animal ,Mice, Transgenic ,medicine.disease_cause ,Green fluorescent protein ,Transcriptome ,03 medical and health sciences ,Mammary Glands, Animal ,0302 clinical medicine ,Lactation ,medicine ,Animals ,Promoter Regions, Genetic ,Hyperplasia ,biology ,Myoepithelial cell ,Mammary Neoplasms, Experimental ,Epithelial Cells ,Cell Biology ,Phenotype ,Cell Transformation, Neoplastic ,030104 developmental biology ,medicine.anatomical_structure ,Phenobarbital ,030220 oncology & carcinogenesis ,Cancer research ,biology.protein ,Female ,Whey Acidic Protein ,Carcinogenesis ,Signal Transduction - Abstract
Hyperactivated Notch signalling has been implicated in breast cancer, but how elevated levels of Notch signalling contribute to mammary dysplasia and tumorigenesis is not fully understood. In this study, we express an activated form of Notch1 in the mouse mammary luminal lineage and analyse the consequences for tumour formation and the transcriptomic landscape in the luminal lineage. Simultaneous conditional activation of a Notch1 intracellular domain (Notch1 ICD) and EGFP in the luminal lineage was achieved by removal of a stop cassette by CRE-recombinase expression from the whey acidic protein (WAP) promoter. Mice in which Notch1 ICD was activated in the luminal lineage (WAP-CRE;R26-N1ICD mice) exhibit ductal hyperplasia after lactation with an increase in branching frequency and in the number of side-branch ends in the ductal tree. A subset of the mice developed mammary tumours and the majority of the tumour cells expressed EGFP (as a proxy for Notch1 ICD), indicating that the tumours originate from the Notch1 ICD-expressing cells. Single-cell transcriptome analysis of the EGFP-positive mammary cells identified six subtypes of luminal cells. The same six subtypes were found in control mice (WAP-CRE;R26-tdTomato mice expressing the tdTomato reporter from WAP-CRE-mediated activation), but the proportion of cells in the various subtypes differed between the WAP-CRE;R26-N1ICD and control WAP-CRE;R26-tdTomato mice. In conclusion, we show that Notch1 ICD expression in the luminal lineage produces a ductal hyperplasia and branching phenotype accompanied by altered luminal cell subtype partitioning.
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- 2020
47. The 1p36 Tumor Suppressor KIF 1Bβ Is Required for Calcineurin Activation, Controlling Mitochondrial Fission and Apoptosis
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John Inge Johnsen, Olga Surova, Susanne Schlisio, Erik Smedler, Zhi Xiong Chen, Per Uhlén, Karin Wallis, Rajappa S. Kenchappa, Shuijie Li, Tommy Martinsson, Stuart M. Fell, Per Kogner, and Ulf Hellman
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0301 basic medicine ,Phosphatase ,Cell Biology ,Biology ,medicine.disease ,General Biochemistry, Genetics and Molecular Biology ,Cell biology ,Dephosphorylation ,Calcineurin ,03 medical and health sciences ,030104 developmental biology ,Biochemistry ,Apoptosis ,Neuroblastoma ,medicine ,Phosphorylation ,Mitochondrial fission ,Signal transduction ,Molecular Biology ,Developmental Biology - Abstract
KIF1Bβ is a candidate 1p36 tumor suppressor that regulates apoptosis in the developing sympathetic nervous system. We found that KIF1Bβ activates the Ca(2+)-dependent phosphatase calcineurin (CN) by stabilizing the CN-calmodulin complex, relieving enzymatic autoinhibition and enabling CN substrate recognition. CN is the key mediator of cellular responses to Ca(2+) signals and its deregulation is implicated in cancer, cardiac, neurodegenerative, and immune disease. We show that KIF1Bβ affects mitochondrial dynamics through CN-dependent dephosphorylation of Dynamin-related protein 1 (DRP1), causing mitochondrial fission and apoptosis. Furthermore, KIF1Bβ actuates recognition of all known CN substrates, implying a general mechanism for KIF1Bβ in Ca(2+) signaling and how Ca(2+)-dependent signaling is executed by CN. Pathogenic KIF1Bβ mutations previously identified in neuroblastomas and pheochromocytomas all fail to activate CN or stimulate DRP1 dephosphorylation. Importantly, KIF1Bβ and DRP1 are silenced in 1p36 hemizygous-deleted neuroblastomas, indicating that deregulation of calcineurin and mitochondrial dynamics contributes to high-risk and poor-prognosis neuroblastoma.
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- 2016
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48. Spinal cord injury in zebrafish induced by near-infrared femtosecond laser pulses
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Sandra Ceccatelli, Sara Hultin, Stefan Spulber, Per Uhlén, Claes Hultling, Ivar Dehnisch Ellström, and Nils Norlin
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0301 basic medicine ,Microsurgery ,Materials science ,Laser pumping ,Neurosurgical Procedures ,law.invention ,03 medical and health sciences ,0302 clinical medicine ,law ,medicine ,Animals ,Zebrafish ,Spinal cord injury ,Spinal Cord Injuries ,Microscopy, Confocal ,biology ,Pulse (signal processing) ,General Neuroscience ,Lasers ,biology.organism_classification ,Spinal cord ,Laser ,medicine.disease ,Light intensity ,Disease Models, Animal ,030104 developmental biology ,medicine.anatomical_structure ,Larva ,Femtosecond ,030217 neurology & neurosurgery ,Locomotion ,Biomedical engineering - Abstract
Background The spinal cord is composed of a large number of cells that interact to allow the organism to function. To perform detail studies of cellular processes involved in spinal cord injury (SCI), one must use repeatable and specific methods to target and injure restricted areas of the spinal cord. New method We propose a robust method to induce SCI in zebrafish by laser light. With a 2-photon microscope equipped with a femtosecond near-infrared pump laser, we explored the effects of laser beam exposure time, area, and intensity to induce precise and repeatable SCI with minimized collateral damage to neighboring cells. Results Through behavioral studies in zebrafish larvae, we assessed the functional outcome of intensive laser light directed at the spinal cord. Our experiments revealed that a laser pulse with wavelength 800 nm, duration 2.6 ms, and light intensity 390 mW was sufficient to induce controlled cell death in a single cell or a spinal cord segment. Collateral damage was observed if cells were exposed to laser pulses exceeding 470 mW. With these settings, we could induce precise and repeatable SCI in zebrafish larvae, resulting in loss of motor and sensory function. Comparison with existing method(s) Our method offers a simple and more controlled setting to induce SCI in zebrafish. We describe how the near-infrared femtosecond laser should be adjusted for achieving optimal results with minimal collateral damage. Conclusions We present a precise and robust method for inducing SCI in zebrafish with single-cell resolution using femtosecond near-infrared laser pulses.
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- 2018
49. The T-type Ca
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Paola, Rebellato, Dagmara, Kaczynska, Shigeaki, Kanatani, Ibrahim Al, Rayyes, Songbai, Zhang, Carlos, Villaescusa, Anna, Falk, Ernest, Arenas, Ola, Hermanson, Lauri, Louhivuori, and Per, Uhlén
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Cerebral Cortex ,Mice, Inbred C57BL ,Mice, Knockout ,Calcium Channels, T-Type ,Neural Stem Cells ,Caspase 3 ,Lateral Ventricles ,Neuroepithelial Cells ,Animals ,Gene Expression Regulation, Developmental ,Cell Differentiation ,Calcium Signaling - Abstract
Here we report that the low-voltage-dependent T-type calcium (Ca
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- 2018
50. Single cell analysis of autism patient with bi-allelic NRXN1-alpha deletion reveals skewed fate choice in neural progenitors and impaired neuronal functionality
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Mohsen Moslem, Loora Laan, Sergiy V. Korol, Anna Falk, Britt-Marie Anderlid, Rebecca Morse, Matti Lam, Niklas Dahl, Robin Pronk, Per Uhlén, Patrick F. Sullivan, Elias Uhlin, Harriet Ronnholm, Julien Bryois, Ivar Dehnisch Ellström, Malin Kele, Jessica Olive, and Lauri Louhivuori
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0301 basic medicine ,Neurogenesis ,Cellular differentiation ,Induced Pluripotent Stem Cells ,Neurexin ,Action Potentials ,Nerve Tissue Proteins ,Biology ,03 medical and health sciences ,0302 clinical medicine ,Neural Stem Cells ,medicine ,Humans ,Autistic Disorder ,Autism spectrum disorder ,Single cell RNA sequencing ,Progenitor cell ,Induced pluripotent stem cell ,Neural Cell Adhesion Molecules ,Neurexin-1 alpha ,Alleles ,Calcium-Binding Proteins ,Neurosciences ,Cell Differentiation ,Cell Biology ,medicine.disease ,Neural stem cell ,Disease modeling ,030104 developmental biology ,Neural development ,030220 oncology & carcinogenesis ,Autism ,Single-Cell Analysis ,Neuroscience ,Gene Deletion ,Neurovetenskaper - Abstract
We generated human iPS derived neural stem cells and differentiated cells from healthy control individuals and an individual with autism spectrum disorder carrying bi-allelic NRXN1-alpha deletion. We investigated the expression of NRXN1-alpha during neural induction and neural differentiation and observed a pivotal role for NRXN1-alpha during early neural induction and neuronal differentiation. Single cell RNA-seq pinpointed neural stem cells carrying NRXN1-alpha deletion shifting towards radial glia-like cell identity and revealed higher proportion of differentiated astroglia. Furthermore, neuronal cells carrying NRXN1-alpha deletion were identified as immature by single cell RNA-seq analysis, displayed significant depression in calcium signaling activity and presented impaired maturation action potential profile in neurons investigated with electrophysiology. Our observations propose NRXN1-alpha plays an important role for the efficient establishment of neural stem cells, in neuronal differentiation and in maturation of functional excitatory neuronal cells.
- Published
- 2019
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