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https://www.readbyqxmd.com/read/28550025/mineralocorticoid-receptor-null-mice-informing-cell-type-specific-roles
#1
Timothy James Cole, Morag Young
The mineralocorticoid receptor (MR) mediates the actions of two important adrenal corticosteroid hormones, aldosterone and cortisol. The cell signalling roles of the MR in vivo have expanded enormously since the cloning of human MR gene 30 years ago and the first MR gene knockout in mice nearly 20 years ago. Complete ablation of the MR revealed important roles postnatally for regulation of kidney epithelial functions, with MR-null mice dying 1-2 weeks postnatally from renal salt wasting and hyperkalaemia, with elevated plasma renin and aldosterone...
May 26, 2017: Journal of Endocrinology
https://www.readbyqxmd.com/read/28549095/klf4-plays-an-essential-role-in-corneal-epithelial-homeostasis-by-promoting-epithelial-cell-fate-and-suppressing-epithelial-mesenchymal-transition
#2
Anil Tiwari, Chelsea L Loughner, Sudha Swamynathan, Shivalingappa K Swamynathan
Purpose: The purpose of this study was to test the hypothesis that KLF4 promotes corneal epithelial (CE) cell fate by suppressing the epithelial-mesenchymal transition (EMT), using spatiotemporally regulated CE-specific ablation of Klf4 in Klf4Δ/ΔCE (Klf4LoxP/LoxP/Krt12rtTA/rtTA/Tet-O-Cre) mice. Methods: CE-specific ablation of Klf4 was achieved by feeding Klf4Δ/ΔCE mice with doxycycline chow. The wild-type (WT; normal chow-fed littermates) and the Klf4Δ/ΔCE histology was compared by hematoxylin and eosin-stained sections; EMT marker expression was quantified by quantitative PCR, immunoblots, and immunofluorescent staining; and wound healing rate was measured by CE debridement using Algerbrush...
May 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/28511701/easi-crispr-a-robust-method-for-one-step-generation-of-mice-carrying-conditional-and-insertion-alleles-using-long-ssdna-donors-and-crispr-ribonucleoproteins
#3
Rolen M Quadros, Hiromi Miura, Donald W Harms, Hisako Akatsuka, Takehito Sato, Tomomi Aida, Ronald Redder, Guy P Richardson, Yutaka Inagaki, Daisuke Sakai, Shannon M Buckley, Parthasarathy Seshacharyulu, Surinder K Batra, Mark A Behlke, Sarah A Zeiner, Ashley M Jacobi, Yayoi Izu, Wallace B Thoreson, Lisa D Urness, Suzanne L Mansour, Masato Ohtsuka, Channabasavaiah B Gurumurthy
BACKGROUND: Conditional knockout mice and transgenic mice expressing recombinases, reporters, and inducible transcriptional activators are key for many genetic studies and comprise over 90% of mouse models created. Conditional knockout mice are generated using labor-intensive methods of homologous recombination in embryonic stem cells and are available for only ~25% of all mouse genes. Transgenic mice generated by random genomic insertion approaches pose problems of unreliable expression, and thus there is a need for targeted-insertion models...
May 17, 2017: Genome Biology
https://www.readbyqxmd.com/read/28498888/cardiomyocyte-specific-loss-of-mitochondrial-p32-c1qbp-causes-cardiomyopathy-and-activates-stress-responses
#4
Toshiro Saito, Takeshi Uchiumi, Mikako Yagi, Rie Amamoto, Daiki Setoyama, Yuichi Matsushima, Dongchon Kang
Aims: Mitochondria are important organelles, dedicated to energy production. Mitochondrial p32/C1qbp, which functions as an RNA and protein chaperone, interacts with mitochondrial mRNA and is indispensable for mitochondrial function through its regulation of mitochondrial translation in cultured cell lines. However, the precise role of p32/C1qbp in vivo is poorly understood because of embryonic lethality in the systemic p32-deficient mouse. The goal of this study was to examine the physiological function of mitochondrial p32/C1qbp in the heart...
May 11, 2017: Cardiovascular Research
https://www.readbyqxmd.com/read/28494939/cardiosphere-derived-cells-require-endoglin-for-paracrine-mediated-angiogenesis
#5
Rachael E Redgrave, Simon Tual-Chalot, Benjamin J Davison, Esha Singh, Darroch Hall, Muhammad M Amirrasouli, Derek Gilchrist, Alexander Medvinsky, Helen M Arthur
Clinical trials of stem cell therapy to treat ischemic heart disease primarily use heterogeneous stem cell populations. Small benefits occur via paracrine mechanisms that include stimulating angiogenesis, and increased understanding of these mechanisms would help to improve patient outcomes. Cardiosphere-derived-cells (CDCs) are an example of these heterogeneous stem cell populations, cultured from cardiac tissue. CDCs express endoglin, a co-receptor that binds specific transforming growth factor β (TGFβ) family ligands, including bone morphogenetic protein 9 (BMP9)...
May 9, 2017: Stem Cell Reports
https://www.readbyqxmd.com/read/28469183/coaxial-3d-bioprinting-of-self-assembled-multicellular-heterogeneous-tumor-fibers
#6
Xingliang Dai, Libiao Liu, Jia Ouyang, Xinda Li, Xinzhi Zhang, Qing Lan, Tao Xu
Three-dimensional (3D) bioprinting of living structures with cell-laden biomaterials has been achieved in vitro, however, some cell-cell interactions are limited by the existing hydrogel. To better mimic tumor microenvironment, self-assembled multicellular heterogeneous brain tumor fibers have been fabricated by a custom-made coaxial extrusion 3D bioprinting system, with high viability, proliferative activity and efficient tumor-stromal interactions. Therein, in order to further verify the sufficient interactions between tumor cells and stroma MSCs, CRE-LOXP switch gene system which contained GSCs transfected with "LOXP-STOP-LOXP-RFP" genes and MSCs transfected with "CRE recombinase" gene was used...
May 3, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28462074/hypothalamic-growth-hormone-receptor-ghr-%C3%A2-controls-hepatic-glucose-production-in-nutrient-sensing-leptin-receptor-leprb-expressing-neurons
#7
Gillian Cady, Taylor Landeryou, Michael Garratt, John J Kopchick, Nathan Qi, David Garcia-Galiano, Carol F Elias, Martin G Myers, Richard A Miller, Darleen A Sandoval, Marianna Sadagurski
OBJECTIVE: The GH/IGF-1 axis has important roles in growth and metabolism. GH and GH receptor (GHR) are active in the central nervous system (CNS) and are crucial in regulating several aspects of metabolism. In the hypothalamus, there is a high abundance of GH-responsive cells, but the role of GH signaling in hypothalamic neurons is unknown. Previous work has demonstrated that the Ghr gene is highly expressed in LepRb neurons. Given that leptin is a key regulator of energy balance by acting on leptin receptor (LepRb)-expressing neurons, we tested the hypothesis that LepRb neurons represent an important site for GHR signaling to control body homeostasis...
May 2017: Molecular Metabolism
https://www.readbyqxmd.com/read/28449847/loss-of-wdfy3-ameliorates-severity-of-serum-transfer-induced-arthritis-independently-of-autophagy
#8
Dennis J Wu, Iannis E Adamopoulos
WDFY3 is a master regulator of selective autophagy that we recently showed to interact with TRAF6 and augment RANKL-induced osteoclastogenesis in vitro and in vivo via the NF-κB pathway. Since the NF-κB pathway plays a major role in inflammation herein, we investigate the role of WDFY3 in an arthritis animal model. Our data show that WDFY3 conditional knockout mice (Wdfy3(loxP/loxP)-LysM-Cre+) were protected in the K/BxN serum transfer-induced arthritis animal model. These effects were independent of alterations in starvation-induced autophagy as evidenced by Western blot analysis of the autophagy marker LC3, autophagosome formation in osteoclast precursors and lysosome formation in osteoclasts derived from WDFY3-cKO mice compared to controls...
April 22, 2017: Cellular Immunology
https://www.readbyqxmd.com/read/28446024/enhanced-vector-design-for-cancer-gene-therapy-with-hierarchical-enhancement-of-therapeutic-transgene-expression
#9
Maria B Kostina, Alexader V Sass, Elena A Stukacheva, Igor V Korobko, Eugeny D Sverdlov
A set of vectors for Cre recombinase-dependent expression of hybrid suicidal FCU1 transgene was constructed which included two-plasmid system where FCU1 and Cre transgenes reside in separate vectors, and single-plasmid variants when a single plasmid bears both transgenes. To improve safety profile and specificity in cancer gene therapy applications, as well as to assure stable propagation of plasmids in bacterial cells, Cre/LoxP system components were optimized. Bicistronic vector with Cre expression cassette placed between LoxP sites unidirectionally with FCU1 cDNA resulted in higher therapeutic efficiency compared to double-plasmid system in enzyme-prodrug suicide cancer gene therapy scheme...
April 27, 2017: Human Gene Therapy Methods
https://www.readbyqxmd.com/read/28433771/a-versatile-tumor-gene-deletion-system-reveals-a-crucial-role-for-fgfr1-in-breast-cancer-metastasis
#10
Wei Wang, Yanling Meng, Bingning Dong, Jie Dong, Michael M Ittmann, Chad J Creighton, Yang Lu, Hong Zhang, Tao Shen, Jianghua Wang, David R Rowley, Yi Li, Fengju Chen, David D Moore, Feng Yang
RCAS avian viruses have been used to deliver oncogene expression and induce tumors in transgenic mice expressing the virus receptor TVA. Here we report the generation and characterization of a novel RCAS-Cre-IRES-PyMT (RCI-PyMT) virus designed to specifically knockout genes of interest in tumors generated in appropriate mutant mouse hosts. FGF receptor 1 (FGFR1) is a gene that is amplified in human breast cancer, but there have been no definitive studies on its function in mammary tumorigenesis, progression, and metastasis in vivo in spontaneous tumors in mice...
May 2017: Neoplasia: An International Journal for Oncology Research
https://www.readbyqxmd.com/read/28420716/a-men1-pancreatic-neuroendocrine-tumour-mouse-model-under-temporal-control
#11
Kate E Lines, Roeland P Vas Nunes, Morten Frost, Christopher J Yates, Mark Stevenson, Rajesh Thakker
Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant disorder characterised by occurrence of parathyroid tumours, and neuroendocrine tumours (NETs) of the pancreatic islets and anterior pituitary. The MEN1 gene, encoding menin, is a tumour suppressor, but its precise role in initiating in vivo tumourigenesis remains to be elucidated. The availability of a temporally controlled conditional MEN1 mouse model would greatly facilitate the study of such early tumourigenic events, and overcome the limitations of other MEN1 knockout models, in which menin is lost from conception, or tumour development occurs asynchronously...
April 18, 2017: Endocrine Connections
https://www.readbyqxmd.com/read/28416649/optic-nerve-astrocyte-reactivity-protects-function-in-experimental-glaucoma-and-other-nerve-injuries
#12
Daniel Sun, Sara Moore, Tatjana C Jakobs
Reactive remodeling of optic nerve head astrocytes is consistently observed in glaucoma and other optic nerve injuries. However, it is unknown whether this reactivity is beneficial or harmful for visual function. In this study, we used the Cre recombinase (Cre)-loxP system under regulation of the mouse glial fibrillary acidic protein promoter to knock out the transcription factor signal transducer and activator of transcription 3 (STAT3) from astrocytes and test the effect this has on reactive remodeling, ganglion cell survival, and visual function after experimental glaucoma and nerve crush...
May 1, 2017: Journal of Experimental Medicine
https://www.readbyqxmd.com/read/28407687/cytokeratin-19-promoter-directs-the-expression-of-cre-recombinase-in-various-epithelia-of-transgenic-mice
#13
Gui-Feng Zhao, Shuang Zhao, Jia-Jie Liu, Ji-Cheng Wu, Hao-Yu He, Xiao-Qing Ding, Xue-Wen Yu, Ke-Qiang Huang, Zhi-Jie Li, Hua-Chuan Zheng
Cytokeratin 19 (K19) is expressed in various differentiated cells, including gastric, intestinal and bronchial epithelial cells, and liver duct cells. Here, we generated a transgenic mouse line, K19-Cre, in which the expression of Cre recombinase was controlled by the promoter of K19. To test the tissue distribution and excision activity of Cre recombinase, K19-Cre transgenic mice were bred with Rosa26 reporter strain and a mouse strain that carries PTEN conditional alleles (PTENLoxp/Loxp). At mRNA level, Cre was strongly expressed in the stomach, lung and intestine, while in stomach, lung, and liver at protein level...
March 14, 2017: Oncotarget
https://www.readbyqxmd.com/read/28401685/characterization-of-new-bone-morphogenetic-protein-bmp-2-regulatory-alleles
#14
Tapan A Shah, Youhua Zhu, Nadia N Shaikh, Marie A Harris, Stephen E Harris, Melissa B Rogers
Bone morphogenetic protein 2 (BMP2, HGNC:1069, GeneID: 650) is a classical morphogen; a molecule that acts at a distance and whose concentration influences cell proliferation, differentiation, and apoptosis. Key events requiring precise Bmp2 regulation include heart specification and morphogenesis and neural development. In mesenchymal cells, the concentration of BMP2 influences myogenesis, adipogenesis, chondrogenesis, and osteogenesis. Because the amount, timing, and location of BMP2 synthesis influence pattern formation and organogenesis, the mechanisms that regulate Bmp2 are crucial...
April 12, 2017: Genesis: the Journal of Genetics and Development
https://www.readbyqxmd.com/read/28401157/engineering-of-systematic-elimination-of-a-targeted-chromosome-in-human-cells
#15
Hiroshi Sato, Hiroki Kato, Haruyoshi Yamaza, Keiji Masuda, Huong Thi Nguyen Nguyen, Thanh Thi Mai Pham, Xu Han, Yuta Hirofuji, Kazuaki Nonaka
Embryonic trisomy leads to abortion or congenital genetic disorders in humans. The most common autosomal chromosome abnormalities are trisomy of chromosomes 13, 18, and 21. Although alteration of gene dosage is thought to contribute to disorders caused by extra copies of chromosomes, genes associated with specific disease phenotypes remain unclear. To generate a normal cell from a trisomic cell as a means of etiological analysis or candidate therapy for trisomy syndromes, we developed a system to eliminate a targeted chromosome from human cells...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28389020/seeds-in-the-liver
#16
REVIEW
Hongjie Ji, Yanrong Lu, Yujun Shi
The liver is a crucial organ for homeostasis and has a tremendous self-renewal and regenerative capacity. It has long been believed that the self-renewal and repair of the liver within a given physiological condition or its repopulation in chronic liver diseases, when hepatocyte proliferation is impaired, will primarily be conducted by the proliferating duct cells, termed "oval cells" or hepatic progenitor cells (HPCs). In addition, numerous studies have revealed that HPCs are the initial tumor cells of liver cancer under certain micro-environments...
April 4, 2017: Acta Histochemica
https://www.readbyqxmd.com/read/28387239/survival-of-human-embryonic-stem-cells-implanted-in-the-guinea-pig-auditory-epithelium
#17
Min Young Lee, Sandra Hackelberg, Kari L Green, Kelly G Lunghamer, Takaomi Kurioka, Benjamin R Loomis, Donald L Swiderski, R Keith Duncan, Yehoash Raphael
Hair cells in the mature cochlea cannot spontaneously regenerate. One potential approach for restoring hair cells is stem cell therapy. However, when cells are transplanted into scala media (SM) of the cochlea, they promptly die due to the high potassium concentration. We previously described a method for conditioning the SM to make it more hospitable to implanted cells and showed that HeLa cells could survive for up to a week using this method. Here, we evaluated the survival of human embryonic stem cells (hESC) constitutively expressing GFP (H9 Cre-LoxP) in deaf guinea pig cochleae that were pre-conditioned to reduce potassium levels...
April 7, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28384353/generation-and-characterization-of-epoxide-hydrolase-3-ephx3-deficient-mice
#18
Samantha L Hoopes, Artiom Gruzdev, Matthew L Edin, Joan P Graves, J Alyce Bradbury, Gordon P Flake, Fred B Lih, Laura M DeGraff, Darryl C Zeldin
Cytochrome P450 (CYP) epoxygenases metabolize arachidonic acid into epoxyeicosatrienoic acids (EETs), which play an important role in blood pressure regulation, protection against ischemia-reperfusion injury, angiogenesis, and inflammation. Epoxide hydrolases metabolize EETs to their corresponding diols (dihydroxyeicosatrienoic acids; DHETs) which are biologically less active. Microsomal epoxide hydrolase (EPHX1, mEH) and soluble epoxide hydrolase (EPHX2, sEH) were identified >30 years ago and are capable of hydrolyzing EETs to DHETs...
2017: PloS One
https://www.readbyqxmd.com/read/28379340/liver-nf-%C3%AE%C2%BAb-inducing-kinase-nik-promotes-liver-steatosis-and-glucose-counterregulation-in-male-mice-with-obesity
#19
Yan Liu, Liang Sheng, Yi Xiong, Hong Shen, Yong Liu, Liangyou Rui
Obesity is associated with chronic inflammation and liver steatoses. Numerous proinflammatory cytokines have been reported to regulate liver glucose and lipid metabolism, thus contributing to the pathogenesis of liver steatosis and/or metabolic dysfunction. NIK is stimulated by many cytokines and mediates activation of the noncanonical NF-κB pathway. We previously reported that liver NIK is aberrantly activated in obesity; inactivation of NIK by overexpressing dominant negative NIK(KA) suppresses hepatic glucose production...
April 3, 2017: Endocrinology
https://www.readbyqxmd.com/read/28378505/prolactin-receptors-in-rip-cre-cells-but-not-in-agrp-neurons-are-involved-in-energy-homeostasis
#20
S R Ladyman, M A MacLeod, Z Khant Aung, P Knowles, H R Phillipps, R S E Brown, D R Grattan
Among its many functions, prolactin has been implicated in energy homeostasis, particularly during pregnancy and lactation. The arcuate nucleus is a key site in the regulation of energy balance. The aim of this study was to examine whether arcuate nucleus neuronal populations involved in energy homeostasis are prolactin responsive and if they can mediate the effects of prolactin on energy homeostasis. To determine if Agrp neurons or Rip-Cre neurons are prolactin responsive, transgenic mice expressing the reporter td-tomato in Agrp neurons (td-tomato/AgRP-Cre) or Rip-Cre neurons (td-tomato/Rip-Cre) were treated with prolactin and perfused 45 minutes later...
April 4, 2017: Journal of Neuroendocrinology
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