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https://www.readbyqxmd.com/read/28077715/excitatory-hindbrain-forebrain-communication-is-required-for-cisplatin-induced-anorexia-and-weight-loss
#1
Amber L Alhadeff, Ruby A Holland, Huiyuan Zheng, Linda Rinaman, Harvey J Grill, Bart C De Jonghe
: Cisplatin chemotherapy is commonly used to treat cancer despite severe energy balance side effects. In rats, cisplatin activates nucleus tractus solitarius (NTS) projections to the lateral parabrachial nucleus (lPBN) and calcitonin-gene related peptide (CGRP) projections from the lPBN to the central nucleus of the amygdala (CeA). We demonstrated previously that CeA glutamate receptor signaling mediates cisplatin-induced anorexia and body weight loss. Here, we used neuroanatomical tracing, immunofluorescence, and confocal imaging to demonstrate that virtually all NTS→lPBN and lPBN→CeA CGRP projections coexpress vesicular glutamate transporter 2 (VGLUT2), providing evidence that excitatory projections mediate cisplatin-induced energy balance dysregulation...
January 11, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28076846/effect-of-trp53-gene-deficiency-on-brain-injury-after-neonatal-hypoxia-ischemia
#2
Ana A Baburamani, Kristina S Sobotka, Regina Vontell, Carina Mallard, Veena G Supramaniam, Claire Thornton, Henrik Hagberg
Hypoxia-ischemia (HI) can result in permanent life-long injuries such as motor and cognitive deficits. In response to cellular stressors such as hypoxia, tumor suppressor protein p53 is activated, potently initiating apoptosis and promoting Bax-dependent mitochondrial outer membrane permeabilization. The aim of this study was to investigate the effect of Trp53 genetic inhibition on injury development in the immature brain following HI. HI (50 min or 60 min) was induced at postnatal day 9 (PND9) in Trp53 heterozygote (het) and wild type (WT) mice...
January 5, 2017: Oncotarget
https://www.readbyqxmd.com/read/28071976/precise-flexible-and-affordable-gene-stacking-for-crop-improvement
#3
Weiqiang Chen, David W Ow
Addendum to: Hou L, Yau YY, Wei J, Han Z, Dong Z, Ow DW. An open-source system for in planta gene stacking by Bxb1 and Cre recombinases. Mol Plant, 2014; 7(12):1756-1765. doi:10.1093/mp/ssu107; Li R, Han Z, Hou L, Kaur G, Yin Q, Ow DW. Method for Biolistic Site-Specific Integration in Plants Catalyzed by Bxb1 Integrase. In: Murata M, editor. Chromosome and Genomic Engineering in Plants: Methods and Protocols, Methods in Molecular Biology. 2016; p.15-30; and Chen W, Ow DW. Protocol for In Vitro Stacked Molecules Compatible with In Vivo Recombinase-Mediated Gene Stacking...
January 10, 2017: Bioengineered
https://www.readbyqxmd.com/read/28063344/reducing-persistent-polyomavirus-infection-increases-functionality-of-virus-specific-memory-cd8-t-cells
#4
Qingsong Qin, Matthew Lauver, Saumya Maru, Eugene Lin, Aron E Lukacher
Mouse polyomavirus (MuPyV) causes a smoldering persistent infection in immunocompetent mice. To lower MuPyV infection in acutely and persistently infected mice, and study the impact of a temporal reduction in viral loads on the memory CD8 T cell response, we created a recombinant MuPyV in which a loxP sequence was inserted into the A2 strain genome upstream of the early promoter and another loxP sequence was inserted in cis into the intron shared by all three T antigens. Using mice transgenic for tamoxifen-inducible Cre recombinase, we demonstrated that reduction in MuPyV load during persistent infection was associated with differentiation of virus-specific CD8 T cells having a superior recall response...
January 4, 2017: Virology
https://www.readbyqxmd.com/read/28062693/pu-1-regulates-ig-light-chain-transcription-and-rearrangement-in-pre-b-cells-during-b-cell-development
#5
Carolina R Batista, Stephen K H Li, Li S Xu, Lauren A Solomon, Rodney P DeKoter
B cell development and Ig rearrangement are governed by cell type- and developmental stage-specific transcription factors. PU.1 and Spi-B are E26-transformation-specific transcription factors that are critical for B cell differentiation. To determine whether PU.1 and Spi-B are required for B cell development in the bone marrow, Spi1 (encoding PU.1) was conditionally deleted in B cells by Cre recombinase under control of the Mb1 gene in Spib (encoding Spi-B)-deficient mice. Combined deletion of Spi1 and Spib resulted in a lack of mature B cells in the spleen and a block in B cell development in the bone marrow at the small pre-B cell stage...
January 6, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28060349/co-localization-of-cell-lineage-markers-and-the-tomato-signal
#6
Yan Jing, Robert J Hinton, Kevin S Chan, Jian Q Feng
The cell lineage tracing system has been used predominantly in developmental biology studies. The use of Cre recombinase allows for the activation of the reporter in a specific cell line and all progeny. Here, we used the cell lineage tracing technique to demonstrate that chondrocytes directly transform into osteoblasts and osteocytes during long bone and mandibular condyle development using two kinds of Cre, Col10a1-Cre and Aggrecan-Cre(ERT2) (Agg-Cre(ERT2)), crossed with Rosa26(tdTomato). Both Col10 and aggrecan are well-recognized markers for chondrocytes...
December 28, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28043906/heterotrimeric-g-stimulatory-protein-alpha-subunit-is-required-for-intestinal-smooth-muscle-contraction-in-mice
#7
Xiaoteng Qin, Shangming Liu, Qiulun Lu, Meng Zhang, Xiuxin Jiang, Sanyuan Hu, Jingxin Li, Cheng Zhang, Jiangang Gao, Min-Sheng Zhu, Yun Zhang, Wencheng Zhang
BACKGROUND & AIMS: The alpha subunit of the heterotrimeric G stimulatory protein (Gsa), encoded by the guanine nucleotide binding protein, alpha stimulating gene (Gnas, in mice), is expressed ubiquitously and mediates receptor-stimulated production of cyclic adenosine monophosphate (cAMP) and activation of the protein kinase A signaling pathway. We investigated the roles of Gsa in vivo in smooth muscle cells of mice. METHODS: We performed studies of mice with Cre recombinase-mediated disruption of Gnas in smooth muscle cells (Gsa(SMKO) and SM22-CreER(T2), induced in adult mice by tamoxifen)...
December 30, 2016: Gastroenterology
https://www.readbyqxmd.com/read/28039299/hyperactivation-of-the-nlrp3-inflammasome-in-myeloid-cells-leads-to-severe-organ-damage-in-experimental-lupus
#8
Ailing Lu, Hua Li, Junling Niu, Shuxian Wu, Guang Xue, Xiaomin Yao, Qiuhong Guo, Nianhong Wan, Paride Abliz, Guiwen Yang, Liguo An, Guangxun Meng
Systemic lupus erythematosus (SLE) is an autoimmune syndrome associated with severe organ damage resulting from the activation of immune cells. Recently, a role for caspase-1 in murine lupus was described, indicating an involvement of inflammasomes in the development of SLE. Among multiple inflammasomes identified, the NLRP3 inflammasome was connected to diverse diseases, including autoimmune encephalomyelitis. However, the function of NLRP3 in SLE development remains elusive. In this study, we explored the role of NLRP3 in the development of SLE using the pristane-induced experimental lupus model...
December 30, 2016: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28029195/expression-of-tamoxifen-inducible-cre-recombinase-in-lcn5-creer-t2-transgenic-mouse-caput-epididymis
#9
Juan Xu, Guangxin Yao, Yanfei Ru, Shengsong Xie
The epididymis, which connects the testis to vas deferens, plays a crucial role regulating sperm maturation and fertilization. Here, a tamoxifen-inducible CreER(T2) recombinase transgenic mouse was generated to study the function of genes in the caput epididymis using the Cre/LoxP system, which is driven by the 1.8-kb Lcn5 promoter (Lcn5-CreER(T2) ). Both CRE recombinase and ER(T2) mRNA were specifically expressed in the caput epididymis, beginning at postnatal Day 30 and increasing thereafter. Crossing these Lcn5-CreER(T2) transgenic mice with Rosa26;mT/mG reporter mice, which express membrane-bound GFP (mGFP) only after CRE is active at its genetic locus, resulted in the presence of GFP only in the middle/distal caput epididymis after tamoxifen induction...
December 28, 2016: Molecular Reproduction and Development
https://www.readbyqxmd.com/read/28028193/smooth-muscle-endothelin-b-receptors-regulate-blood-pressure-but-not-vascular-function-or-neointimal-remodeling
#10
Eileen Miller, Alicja Czopek, Karolina M Duthie, Nicholas S Kirkby, Elisabeth E Fransen van de Putte, Sibylle Christen, Robert A Kimmitt, Rebecca Moorhouse, Raphael F P Castellan, Yuri V Kotelevtsev, Rhoda E Kuc, Anthony P Davenport, Neeraj Dhaun, David J Webb, Patrick W F Hadoke
The role of smooth muscle endothelinB (ETB) receptors in regulating vascular function, blood pressure (BP), and neointimal remodeling has not been established. Selective knockout mice were generated to address the hypothesis that loss of smooth muscle ETB receptors would reduce BP, alter vascular contractility, and inhibit neointimal remodeling. ETB receptors were selectively deleted from smooth muscle by crossing floxed ETB mice with those expressing cre-recombinase controlled by the transgelin promoter. Functional consequences of ETB deletion were assessed using myography...
December 27, 2016: Hypertension
https://www.readbyqxmd.com/read/28024880/embryonic-ablation-of-neuronal-vgf-increases-energy-expenditure-and-reduces-body-weight
#11
Cheng Jiang, Wei-Jye Lin, Masato Sadahiro, Andrew C Shin, Christoph Buettner, Stephen R Salton
Germline ablation of VGF, a secreted neuronal, neuroendocrine, and endocrine peptide precursor, results in lean, hypermetabolic, and infertile adult mice that are resistant to diet-, lesion-, and genetically-induced obesity and diabetes (Hahm et al., 1999, 2002). To assess whether this phenotype is predominantly driven by reduced VGF expression in developing and/or adult neurons, or in peripheral endocrine and neuroendocrine tissues, we generated and analyzed conditional VGF knockout mice, obtained by mating loxP-flanked (floxed) Vgf mice with either pan-neuronal Synapsin-Cre- or forebrain alpha-CaMKII-Cre-recombinase-expressing transgenic mice...
December 20, 2016: Neuropeptides
https://www.readbyqxmd.com/read/28011085/self-excising-cre-mutant-lox-marker-recycling-system-for-multiple-gene-integrations-and-consecutive-gene-deletions-in-aspergillus-oryzae
#12
Silai Zhang, Akihiko Ban, Naoki Ebara, Osamu Mizutani, Mizuki Tanaka, Takahiro Shintani, Katsuya Gomi
In this study, we developed a self-excising Cre/loxP-mediated marker recycling system with mutated lox sequences to introduce a number of biosynthetic genes into Aspergillus oryzae. To construct the self-excising marker cassette, both the selectable marker, the Aspergillus nidulans adeA gene, and the Cre recombinase gene (cre), conditionally expressed by the xylanase-encoding gene promoter, were designed to be located between the mutant lox sequences, lox66 and lox71. However, construction of the plasmid failed, possibly owing to a slight expression of cre downstream of the fungal gene promoter in Escherichia coli...
December 20, 2016: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28009940/an-in-vivo-transfection-system-for-inducible-gene-expression-and-gene-silencing-in-murine-hepatocytes
#13
Eric K Hubner, Christian Lechler, Birgit Kohnke-Ertel, Anne-Flore Zmoos, Julien Sage, Roland M Schmid, Ursula Ehmer
BACKGROUND: Hydrodynamic tail vein injection (HTVI) of transposon-based integration vectors is an established system to stably transfect mouse hepatocytes in vivo that has been successfully employed to study key questions in liver biology and cancer. Refining the vectors for transposon-mediated hepatocyte transfection will further expand the range of applications of this technique in liver research. Here, we present an advanced transposon-based system to manipulate gene expression in hepatocytes in vivo...
December 23, 2016: Journal of Gene Medicine
https://www.readbyqxmd.com/read/28003412/dicer-maintains-the-identity-and-function-of-proprioceptive-sensory-neurons
#14
Sean Michael O'Toole, Monica M Ferrer, Jennifer Mekonnen, Haihan Zhang, Yasuyuki Shima, David R Ladle, Sacha B Nelson
Neuronal cell identity is established during development and must be maintained throughout an animal's life (Fishell and Heintz 2013). Transcription factors critical for establishing neuronal identity can be required for maintaining it (Deneris and Hobert 2014). Post-transcriptional regulation also plays an important role in neuronal differentiation (Bian and Sun 2011), but its role in maintaining cell identity is less established. To better understand how post-transcriptional regulation might contribute to cell identity, we examined the proprioceptive neurons in the dorsal root ganglion (DRG), a highly specialized sensory neuron class, with well established properties that distinguish them from other neurons in the ganglion...
December 21, 2016: Journal of Neurophysiology
https://www.readbyqxmd.com/read/27999416/dermal-delivery-of-constructs-encoding-cre-recombinase-to-induce-skin-tumors-in-pten-loxp-loxp-braf-ca-mice
#15
Marcel A Deken, Ji-Ying Song, Jules Gadiot, Adriaan D Bins, Paula Kroon, Inge Verbrugge, Christian U Blank
Current genetically-engineered mouse melanoma models are often based on Tyr::CreER(T2)-controlled MAPK pathway activation by the BRAF(V600E) mutation and PI3K pathway activation by loss of PTEN. The major drawback of these models is the occurrence of spontaneous tumors caused by leakiness of the Tyr::CreER(T2) system, hampering long-term experiments. To address this problem, we investigated several approaches to optimally provide local delivery of Cre recombinase, including injection of lentiviral particles, DNA tattoo administration and particle-mediated gene transfer, to induce melanomas in Pten(LoxP/LoxP);Braf(CA/+) mice lacking the Tyr::CreER(T2) allele...
December 20, 2016: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27998284/the-glucose-transporter-glut1-is-required-for-erbb2-induced-mammary-tumorigenesis
#16
Elizabeth A Wellberg, Stevi Johnson, Jessica Finlay-Schultz, Andrew S Lewis, Kristina L Terrell, Carol A Sartorius, E Dale Abel, William J Muller, Steven M Anderson
BACKGROUND: Altered tumor cell metabolism is an emerging hallmark of cancer; however, the precise role for glucose in tumor initiation is not known. GLUT1 (SLC2A1) is expressed in breast cancer cells and is likely responsible for avid glucose uptake observed in established tumors. We have shown that GLUT1 was necessary for xenograft tumor formation from primary mammary cells transformed with the polyomavirus middle-T antigen but that it was not necessary for growth after tumors had formed in vivo, suggesting a differential requirement for glucose depending on the stage of tumorigenesis...
December 20, 2016: Breast Cancer Research: BCR
https://www.readbyqxmd.com/read/27997037/descending-projections-from-the-basal-forebrain-to-the-orexin-neurons-in-mice
#17
Lindsay J Agostinelli, Loris L Ferrari, Carrie E Mahoney, Takatoshi Mochizuki, Bradford B Lowell, Elda Arrigoni, Thomas E Scammell
The orexin (hypocretin) neurons play an essential role in promoting arousal, and loss of the orexin neurons results in narcolepsy, a condition characterized by chronic sleepiness and cataplexy. The orexin neurons excite wake-promoting neurons in the basal forebrain (BF), and a reciprocal projection from the BF back to the orexin neurons may help promote arousal and motivation. The BF contains at least three different cell types (cholinergic, glutamatergic, and GABAergic neurons) across its different regions (medial septum, diagonal band, magnocellular preoptic area, and substantia innominata)...
December 20, 2016: Journal of Comparative Neurology
https://www.readbyqxmd.com/read/27983648/cre-fused-with-rvg-peptide-mediates-targeted-genome-editing-in-mouse-brain-cells-in-vivo
#18
Zhiyuan Zou, Zhaolin Sun, Pan Li, Tao Feng, Sen Wu
Cell penetrating peptides (CPPs) are short peptides that can pass through cell membranes. CPPs can facilitate the cellular entry of proteins, macromolecules, nanoparticles and drugs. RVG peptide (RVG hereinafter) is a 29-amino-acid CPP derived from a rabies virus glycoprotein that can cross the blood-brain barrier (BBB) and enter brain cells. However, whether RVG can be used for genome editing in the brain has not been reported. In this work, we combined RVG with Cre recombinase for bacterial expression. The purified RVG-Cre protein cut plasmids in vitro and traversed cell membranes in cultured Neuro2a cells...
December 14, 2016: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27931797/erbb2-is-essential-for-the-growth-of-chemically-induced-skin-tumors-in-mice
#19
Maik Dahlhoff, Sukalp Muzumdar, Matthias Schäfer, Marlon R Schneider
While the epidermal growth factor receptor has established roles in skin carcinogenesis, inflammation, and wound healing, the functions of the structurally related receptor ERBB2 in this tissue remain poorly explored. To assess the functions of ERBB2 in skin homeostasis, tumorigenesis, and wound healing we employed keratin 5-directed, cre recombinase-mediated targeting of Erbb2 alleles in mice. Erbb2(del) mice, lacking ERBB2 specifically in keratinocytes, showed no noticeable spontaneous skin abnormalities...
December 5, 2016: Journal of Investigative Dermatology
https://www.readbyqxmd.com/read/27911376/rapid-and-efficient-generation-of-recombinant-human-pluripotent-stem-cells-by-recombinase-mediated-cassette-exchange-in-the-aavs1-locus
#20
Laura Ordovás, Ruben Boon, Mariaelena Pistoni, Yemiao Chen, Rangarajan Sambathkumar, Nicky Helsen, Jolien Vanhove, Pieter Berckmans, Qing Cai, Kim Vanuytsel, Susanna Raitano, Catherine M Verfaillie
Even with the revolution of gene-targeting technologies led by CRISPR-Cas9, genetic modification of human pluripotent stem cells (hPSCs) is still time consuming. Comparative studies that use recombinant lines with transgenes integrated into safe harbor loci could benefit from approaches that use site-specific targeted recombinases, like Cre or FLPe, which are more rapid and less prone to off-target effects. Such methods have been described, although they do not significantly outperform gene targeting in most aspects...
November 20, 2016: Journal of Visualized Experiments: JoVE
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