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https://www.readbyqxmd.com/read/29329320/atypical-e2f-functions-are-critical-for-pancreas-polyploidization
#1
Ramadhan B Matondo, Eva Moreno, Mathilda J M Toussaint, Peter C J Tooten, Saskia C van Essen, Elsbeth A van Liere, Sameh A Youssef, Laura Bongiovanni, Alain de Bruin
The presence of polyploid cells in the endocrine and exocrine pancreas has been reported for four decades. In rodents, pancreatic polyploidization is initiated after weaning and the number of polyploid cells increases with age. Surprisingly the molecular regulators and biological functions of polyploidization in the pancreas are still unknown. We discovered that atypical E2f activity is essential for polyploidization in the pancreas, using an inducible Cre/LoxP approach in new-born mice to delete ubiquitously the atypical E2f transcription factors, E2f7 and E2f8...
2018: PloS One
https://www.readbyqxmd.com/read/29324843/beta-hpv38-oncoproteins-act-with-a-hit-and-run-mechanism-in-ultraviolet-radiation-induced-skin-carcinogenesis-in-mice
#2
Daniele Viarisio, Karin Müller-Decker, Rosita Accardi, Alexis Robitaille, Matthias Dürst, Katrin Beer, Lars Jansen, Christa Flechtenmacher, Matthias Bozza, Richard Harbottle, Catherine Voegele, Maude Ardin, Jiri Zavadil, Sandra Caldeira, Lutz Gissmann, Massimo Tommasino
Cutaneous beta human papillomavirus (HPV) types are suspected to be involved, together with ultraviolet (UV) radiation, in the development of non-melanoma skin cancer (NMSC). Studies in in vitro and in vivo experimental models have highlighted the transforming properties of beta HPV E6 and E7 oncoproteins. However, epidemiological findings indicate that beta HPV types may be required only at an initial stage of carcinogenesis, and may become dispensable after full establishment of NMSC. Here, we further investigate the potential role of beta HPVs in NMSC using a Cre-loxP-based transgenic (Tg) mouse model that expresses beta HPV38 E6 and E7 oncogenes in the basal layer of the skin epidermis and is highly susceptible to UV-induced carcinogenesis...
January 2018: PLoS Pathogens
https://www.readbyqxmd.com/read/29323723/haplo-insufficiency-of-bcl2-associated-athanogene-3-in-mice-results-in-progressive-left-ventricular-dysfunction-%C3%AE-adrenergic-insensitivity-and-increased-apoptosis
#3
Valerie D Myers, Dhanendra Tomar, Muniswamy Madesh, JuFang Wang, Jianliang Song, Xue-Qian Zhang, Manish K Gupta, Farzaneh G Tahrir, Jennifer Gordon, Joseph M McClung, Christopher D Kontos, Kamel Khalili, Joseph Y Cheung, Arthur M Feldman
Bcl2-associated athanogene 3 (BAG3) is a 575 amino acid protein that is found predominantly in the heart, skeletal muscle and many cancers. Deletions and truncations in BAG3 that result in haplo-insufficiency have been associated with the development of dilated cardiomyopathy. To study the cellular and molecular events attributable to BAG3 haplo-insufficiency we generated a mouse in which one allele of BAG3 was flanked by loxP recombination sites (BAG3fl/+ ). Mice were crossed with α-MHC-Cre mice in order to generate mice with cardiac-specific haplo-insufficiency (cBAG3+/-) and underwent bi-weekly echocardiography to assess their cardiac phenotype...
January 11, 2018: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/29322219/sirt6-mediated-transcriptional-suppression-of-txnip-is-critical-for-pancreatic-beta-cell-function-and-survival-in-mice
#4
Kunhua Qin, Ning Zhang, Zhao Zhang, Michael Nipper, Zhenxin Zhu, Jake Leighton, Kexin Xu, Nicolas Musi, Pei Wang
AIMS/HYPOTHESIS: Better understanding of how genetic and epigenetic components control beta cell differentiation and function is key to the discovery of novel therapeutic approaches to prevent beta cell dysfunction and failure in the progression of type 2 diabetes. Our goal was to elucidate the role of histone deacetylase sirtuin 6 (SIRT6) in beta cell development and homeostasis. METHODS: Sirt6 endocrine progenitor cell conditional knockout and beta cell-specific knockout mice were generated using the Cre-loxP system...
January 10, 2018: Diabetologia
https://www.readbyqxmd.com/read/29286375/inducing-cre-lox-recombination-in-mouse-cerebral-cortex-through-in-utero-electroporation
#5
Katherine M Bland, Zachary O Casey, Christopher J Handwerk, Z Logan Holley, George S Vidal
Cell-autonomous neuronal functions of genes can be revealed by causing loss or gain of function of a gene in a small and sparse population of neurons. To do so requires generating a mosaic in which neurons with loss or gain of function of a gene are surrounded by genetically unperturbed tissue. Here, we combine the Cre-lox recombination system with in utero electroporation in order to generate mosaic brain tissue that can be used to study the cell-autonomous function of genes in neurons. DNA constructs (available through repositories), coding for a fluorescent label and Cre recombinase, are introduced into developing cortical neurons containing genes flanked with loxP sites in the brains of mouse embryos using in utero electroporation...
November 17, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29274742/effect-of-early-embryonic-deletion-of-huntingtin-from-pyramidal-neurons-on-the-development-and-long-term-survival-of-neurons-in-cerebral-cortex-and-striatum
#6
I Dragatsis, P Dietrich, H Ren, Y P Deng, N Del Mar, H B Wang, I M Johnson, K R Jones, A Reiner
We evaluated the impact of early embryonic deletion of huntingtin (htt) from pyramidal neurons on cortical development, cortical neuron survival and motor behavior, using a cre-loxP strategy to inactivate the mouse htt gene (Hdh) in emx1-expressing cell lineages. Western blot confirmed substantial htt reduction in cerebral cortex of these Emx-httKO mice, with residual cortical htt in all likelihood restricted to cortical interneurons of the subpallial lineage and/or vascular endothelial cells. Despite the loss of htt early in development, cortical lamination was normal, as revealed by layer-specific markers...
December 21, 2017: Neurobiology of Disease
https://www.readbyqxmd.com/read/29274472/protein-phosphatase-2ac%C3%AE-gene-knock-out-results-in-cortical-atrophy-through-activating-hippo-cascade-in-neuronal-progenitor-cells
#7
Bo Liu, Li-Hua Sun, Yan-Fei Huang, Li-Jun Guo, Li-Shu Luo
Protein phosphatase 2ACα (PP2ACα), a vital member of the protein phosphatase family, has been studied primarily as a regulator for the development, growth and protein synthesis of a lot of cell types. Dysfunction of PP2ACα protein results in neurodegenerative disease; however, this finding has not been directly confirmed in the mouse model with PP2ACα gene knock-out. Therefore, in this study presented here, we generated the PP2ACα gene knock-out mouse model by the Cre-loxP targeting gene system, with the purpose to directly observe the regulatory role of PP2ACα gene in the development of mouse's cerebral cortex...
December 20, 2017: International Journal of Biochemistry & Cell Biology
https://www.readbyqxmd.com/read/29274400/conditional-mitotimer-reporter-mice-for-assessment-of-mitochondrial-structure-oxidative-stress-and-mitophagy
#8
Rebecca J Wilson, Joshua C Drake, Di Cui, Mei Zhang, Heather M Perry, Jennifer A Kashatus, Christine M Kusminski, Philipp E Scherer, David F Kashatus, Mark D Okusa, Zhen Yan
Assessment of structural and functional changes of mitochondria is vital for biomedical research as mitochondria are the power plants essential for biological processes and tissue/organ functions. Others and we have developed a novel reporter gene, pMitoTimer, which codes for a redox sensitive mitochondrial targeted protein that switches from green fluorescence protein (GFP) to red fluorescent protein (DsRed) when oxidized. It has been shown in transfected cells, transgenic C. elegans and Drosophila m., as well as somatically transfected adult skeletal muscle that this reporter gene allows quantifiable assessment of mitochondrial structure, oxidative stress, and lysosomal targeting of mitochondria-containing autophagosomes...
December 20, 2017: Mitochondrion
https://www.readbyqxmd.com/read/29244772/sphingosine-1-phosphate-receptor-1-expressed-in-myeloid-cells-slows-diet-induced-atherosclerosis-and-protects-against-macrophage-apoptosis-in-ldlr-ko-mice
#9
Leticia Gonzalez, Alexander S Qian, Usama Tahir, Pei Yu, Bernardo L Trigatti
We generated myeloid specific sphingosine-1-phosphate receptor 1 (S1pr1) deficient mice by crossing mice that had myeloid specific expression of Cre recombinase (lyzMCre) with mice having the S1pr1 gene flanked by loxP recombination sites. We transplanted bone marrow from these mice and control lyzMCre mice with intact macrophage S1pr1 gene expression into low-density lipoprotein (LDL) receptor gene (Ldlr) deficient mice. The resulting chimeras were fed a high fat atherogenic diet for nine or twelve weeks and evaluated for atherosclerosis development in the aortic sinus...
December 15, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29243041/cre-loxp-mediated-multicopy-integration-of-the-mevalonate-operon-into-the-genome-of-methylobacterium-extorquens-am1
#10
Wei-Fan Liang, Ming-Yang Sun, Lan-Yu Cui, Chong Zhang, Xin-Hui Xing
Methylobacterium extorquens AM1 is the model strain for methylotrophic bacteria that metabolize methanol as the sole carbon and energy source. Genetically modified M. extorquens AM1 is used as a methylotrophic cell factory (MeCF) for high value-added chemical production. We tested the Cre-loxP recombination system for its ability to mediate multicopy gene integration of the mvt3 operon (mvt3) in M. extorquens AM1. mvt3 controls the expression of the first three enzymes of the mevalonate synthesis pathway. We assayed for Cre-mediated multigene integration by screening for multicopy mutants via their survival in culture with a high kanamycin concentration (600 μg/mL)...
December 15, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/29234326/toll-like-receptor-4-on-both-myeloid-cells-and-dendritic-cells-is-required-for-systemic-inflammation-and-organ-damage-after-hemorrhagic-shock-with-tissue-trauma-in-mice
#11
Kent Zettel, Sebastian Korff, Ruben Zamora, Adrian E Morelli, Sophie Darwiche, Patricia A Loughran, Greg Elson, Limin Shang, Susana Salgado-Pires, Melanie J Scott, Yoram Vodovotz, Timothy R Billiar
Trauma combined with hemorrhagic shock (HS/T) leads to systemic inflammation, which results in organ injury. Toll-like Receptor 4 (TLR4)-signaling activation contributes to the initiation of inflammatory pathways following HS/T but its cell-specific roles in this setting are not known. We assessed the importance of TLR4 on leukocytes of myeloid lineage and dendritic cells (DCs) to the early systemic inflammatory response following HS/T. Mice were subjected to HS/T and 20 inflammatory mediators were measured in plasma followed by Dynamic Bayesian Network (DBN) Analysis...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/29225599/promoter-specificity-and-efficacy-in-conditional-and-inducible-transgenic-targeting-of-lung-macrophages
#12
Alexandra L McCubbrey, Kristen C Allison, Alisa B Lee-Sherick, Claudia V Jakubzick, William J Janssen
Conditional and inducible Cre-loxP systems are used to target gene deletion to specific cell lineages and tissues through promoter-restricted expression of the bacterial DNA recombinase, Cre. Although Cre-loxP systems are widely used to target gene deletion in lung macrophages, limited data are published on the specificity and efficiency of "macrophage targeting" Cre lines. Using R26-stopfl/fl-TdTomato and tetOn-GFP reporter lines, we assessed the specificity and efficiency of four commercially available Cre driver lines that are often considered "macrophage specific...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/29224189/acute-loss-of-adipose-tissue-derived-adiponectin-triggers-immediate-metabolic-deterioration-in-mice
#13
Jonathan Y Xia, Kai Sun, Chelsea Hepler, Alexandra L Ghaben, Rana K Gupta, Yu A An, William L Holland, Thomas S Morley, Andrew C Adams, Ruth Gordillo, Christine M Kusminski, Philipp E Scherer
AIM/HYPOTHESIS: Adiponectin (APN), a circulating hormone secreted by mature adipocytes, has been extensively studied because it has beneficial metabolic effects. While many studies have focused on the congenital loss of APN and its effects on systemic body glucose and lipid metabolism, little is known about the effects triggered by acute loss of APN in the adult mouse. We anticipated that genetically induced acute depletion of APN in adult mice would have a more profound effect on systemic metabolic health than congenital deletion of Adipoq, the gene encoding APN, with its associated potential for adaptive responses that may mask the phenotypes...
December 9, 2017: Diabetologia
https://www.readbyqxmd.com/read/29207258/macrophage-dependent-cytoplasmic-transfer-during-melanoma-invasion-in%C3%A2-vivo
#14
Minna Roh-Johnson, Arish N Shah, Jason A Stonick, Kumud R Poudel, Julia Kargl, Grace H Yang, Julie di Martino, Rafael E Hernandez, Charles E Gast, Luai R Zarour, Susumu Antoku, A McGarry Houghton, Jose Javier Bravo-Cordero, Melissa H Wong, John Condeelis, Cecilia B Moens
Interactions between tumor cells and tumor-associated macrophages play critical roles in the initiation of tumor cell motility. To capture the cellular interactions of the tumor microenvironment with high-resolution imaging, we directly visualized tumor cells and their interactions with macrophages in zebrafish. Live imaging in zebrafish revealed that macrophages are dynamic, yet maintain sustained contact with tumor cells. In addition, the recruitment of macrophages to tumor cells promotes tumor cell dissemination...
December 4, 2017: Developmental Cell
https://www.readbyqxmd.com/read/29205257/ischemic-stroke-activates-the-ve-cadherin-promoter-and-increases-ve-cadherin-expression-in-adult-mice
#15
Akiko Nakano-Doi, Rika Sakuma, Tomohiro Matsuyama, Takayuki Nakagomi
Endothelial cells (ECs) are a key component of the blood-brain barrier (BBB). Healthy ECs in the BBB form inter-endothelial junctions, including adherens junctions (AJs). Under pathological conditions, such as after ischemic stroke, the BBB may be functionally compromised. However, gene and protein expression patterns involving endothelial AJs have not been well studied. Because expression levels of endothelial AJs are considered to be related to BBB functionality, we investigated the expression pattern of a representative endothelial AJ marker, VE-cadherin, in healthy and diseased mice...
November 5, 2017: Histology and Histopathology
https://www.readbyqxmd.com/read/29204649/aav2-mediated-transduction-of-the-mouse-retina-after-optic-nerve-injury
#16
Robert W Nickells, Heather M Schmitt, Margaret E Maes, Cassandra L Schlamp
Purpose: Gene therapy of retinal ganglion cells (RGCs) has promise as a powerful therapeutic for the rescue and regeneration of these cells after optic nerve damage. However, early after damage, RGCs undergo atrophic changes, including gene silencing. It is not known if these changes will deleteriously affect transduction and transgene expression, or if the therapeutic protein can influence reactivation of the endogenous genome. Methods: Double-transgenic mice carrying a Rosa26-(LoxP)-tdTomato reporter, and a mutant allele for the proapoptotic Bax gene were reared...
December 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/29204184/viral-cre-loxp-tools-aid-genome-engineering-in-mammalian-cells
#17
Ranjita Sengupta, Amy Mendenhall, Nandita Sarkar, Chandreyee Mukherjee, Amirali Afshari, Joseph Huang, Biao Lu
Background: Targeted nucleases have transformed genome editing technology, providing more efficient methods to make targeted changes in mammalian genome. In parallel, there is an increasing demand of Cre-LoxP technology for complex genome manipulation such as large deletion, addition, gene fusion and conditional removal of gene sequences at the target site. However, an efficient and easy-to-use Cre-recombinase delivery system remains lacking. Results: We designed and constructed two sets of expression vectors for Cre-recombinase using two highly efficient viral systems, the integrative lentivirus and non-integrative adeno associated virus...
2017: Journal of Biological Engineering
https://www.readbyqxmd.com/read/29203334/sex-differences-in-olfactory-induced-neural-activation-of-the-amygdala
#18
Takefumi Kikusui, Mayu Kajita, Natsumi Otsuka, Tatsuya Hattori, Kanako Kumazawa, Akiyuki Watarai, Miho Nagasawa, Ayumu Inutsuka, Akihiro Yamanaka, Naoki Matsuo, Herbert E Covington, Kazutaka Mogi
Olfactory signals, including the scent of urine, are thought to be processed by specific brain regions, such as the medial amygdala (Me), and regulate sexual behavior in a sex-dependent manner. We aimed to reveal the sex-specific neural circuit from the accessory olfactory bulb (AOB) to Me by using a transgenic mouse. We quantified the long-lasting green fluorescent protein (GFP) expression profile, which was controlled by the c-fos promotor in a sex-dependent manner by the scent of urine. Female urine predominantly activated neurons of the MePD in male mice and the MePV in female mice...
December 1, 2017: Behavioural Brain Research
https://www.readbyqxmd.com/read/29188052/biological-roles-of-hepatocyte-growth-factor-met-signaling-from-genetically-modified-animals
#19
Takashi Kato
Hepatocyte growth factor (HGF) is produced by stromal and mesenchymal cells, and it stimulates epithelial cell proliferation, motility, morphogenesis and angiogenesis in various organs via tyrosine phosphorylation of its cognate receptor, Met. The HGF-Met signaling pathway contributes in a paracrine manner to the development of epithelial organs, exerts regenerative effects on the epithelium, and promotes the regression of fibrosis in numerous organs. Additionally, the HGF-Met signaling pathway is correlated with the biology of cancer types, neurons and immunity...
December 2017: Biomedical Reports
https://www.readbyqxmd.com/read/29180115/deletion-of-girk2-subunit-containing-girk-channels-of-neurons-expressing-dopamine-transporter-decrease-immobility-time-on-forced-swimming-in-mice
#20
Ikutaro Honda, Kimi Araki, Sokichi Honda, Fumio Soeda, Min-Chul Shin, Shogo Misumi, Ken-Ichi Yamamura, Kazuo Takahama
We previously reported that non-narcotic antitussives possessing inhibitory actions on G protein-coupled inwardly rectifying potassium (GIRK) channels have antidepressant-like effects in the forced swimming test in normal and adrenocoticotropic hormone (ACTH) treated rats. Furthermore, the antidepressant-like effects of the antitussives such as tipepidine were blocked by dopamine D1 receptor antagonist, and inhibitory actions on GIRK channels of dopamine neurons may be involved in the antidepressant-like effects of tipepidine...
November 24, 2017: Neuroscience Letters
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