Inducible Transgenic Mouse Models , SpringerLink Inducible transgenic mouse models allow for the activation of and the lac and GAL4 inducible systems. The tetracycline-regulated transgenic models are typically Inducible Gene Expression and Gene Modification in Transgenic two major systems have been successfully used in transgenic mice, i.e., the tetracycline-inducible transgenic mice that express that inducible Cre Introduction to Tet expression systems - The how to take viagra 100mg Jackson Laboratory Learn the basics about how Tet-On and Tet-Off inducible systems Introduction viagra high blood pressure to Tet expression systems. in transgenic mice by a tetracycline Conditional and inducible transgene expression in mice Here we describe a triple transgenic mouse system, which combines the tissue specificity of any Cre-transgenic line with the inducibility of the reverse tetracycline Inducible podocyte-specific gene expression in transgenic Inducible podocyte-specific gene expression in ...
Abstract: : Purpose: To establish a binary inducible transgenic mouse model that can be used to elucidate the roles of growth factors, e.g., FGF-7, on corneal biology. It has been suggested that FGF-7 (KGF, keratinocyte growth factor) serves as a paracrine that modulates the growth of corneal epithelial cell. The present studies are to investigate the effect of excess FGF-7 on corneal epithelial cells in a binary tetracycline inducible transgenic mouse line. Methods: A keratocyte-specific 3.2 kb murine keratocan promotor (Kerapr) has been used to prepare Kerapr-rtTA transgenic (KeraprrtTA/+) mice that constitutively overexpress rtTA (reverse tetracycline transcription activator) in cornea. The KeraprrtTA/+ mice were crossed with tet-OFGF7/FGF7 mice to produce compound heterozygous transgenic (KeraprrtTA/+•tet-OFGF7/+) mice, which were fed doxycycline water (0.5 mg/ml) for one week. The experimental mice were i.p. injected with BrdU (100 µg/g body weight) 2 h prior to sacrifice. The enucleated ...
Despite wide academic and commercial interest in the actions of GLP-1, attempts to identify the cellular targets of GLP-1 are hampered by the lack of specificity of antibodies to GLP1R. Our development of a new transgenic mouse model expressing Cre recombinase driven by the glp1r promoter provides an antibody-independent method for the identification and characterization of live cells expressing glp1r, using floxed fluorescent reporter strains. The results illuminate not only which tissues exhibited glp1r fluorescence but also those that did not.. Establishing definitively that the GLP1R protein is produced by all glp1r-fluorescent cells will be important, because our use of Cre recombinase results in a permanent activation of the fluorescent reporters, even in cells that no longer express the receptor as well as in the progeny of cells that have once expressed glp1r. Where neurones were identified, we were able to confirm expression of GLP1R protein by demonstrating functional responsiveness to ...
Purpose: : To develop a mouse line in which a gene of interest can be removed in keratocytes-specific manner upon induction by doxycycline. Methods: : Two transgenes including Kera3.2-int-rtTA and tet-O-Cre were co-microinjected into the fertilized mouse eggs to create transgenic mice. The Kera3.2-int-rtTA /tet-O-Cre transgenic mice were crossed with reporter Rosa26-LacZ mice to obtain Kera3.2-int-rtTA /tet-O-Cre/Rosa26-LacZ triple transgenic mice. The Cre activity was assessed by the detection of whole mount X-gal staining in corneal stroma of triple transgenic mice after administration of doxycycline via drinking water and chow. Results: : We obtained two independent transgenic founder lines in which the Kera3.2-int-rtTA and tet-O-Cre transgenes were co-segregated at the chromosome level. Administration of doxycycline (Dox) to transgenic Kera3.2-int-rtTA /tet-O-Cre mice did not induce expression of LacZ. The Kera3.2-int-rtTA /tet-O-Cre/Rosa26-LacZ triple transgenic mice showed a little bit ...
TY - JOUR. T1 - Proteomic analysis of Nrf2 deficient transgenic mice reveals cellular defence and lipid metabolism as primary Nrf2-dependent pathways in the liver. AU - Kitteringham, Neil R.. AU - Abdullah, Azman. AU - Walsh, Joanne. AU - Randle, Laura. AU - Jenkins, Rosalind E.. AU - Sison, Rowena. AU - Goldring, Christopher E.P.. AU - Powell, Helen. AU - Sanderson, Christopher. AU - Williams, Samantha. AU - Higgins, Larry. AU - Yamamoto, Masayuki. AU - Hayes, John. AU - Park, B. Kevin. PY - 2010/6/16. Y1 - 2010/6/16. N2 - The transcription factor Nrf2 regulates expression of multiple cellular defence proteins through the antioxidant response element (ARE). Nrf2-deficient mice (Nrf2-/-) are highly susceptible to xenobiotic-mediated toxicity, but the precise molecular basis of enhanced toxicity is unknown. Oligonucleotide array studies suggest that a wide range of gene products is altered constitutively, however no equivalent proteomics analyses have been conducted. To define the range of ...
Am J Pathol. 2012 Feb;180(2):727-37. doi: 10.1016/j.ajpath.2011.10.035. Epub 2011 Dec 7. Research Support, N.I.H., Extramural; Research Support, Non-U.S. Govt
TY - JOUR. T1 - Overexpression of transforming growth factor α in transgenic mice alters nonreproductive, sex-related behavioral differences. T2 - Interaction with gonadal hormones. AU - Hilakivi-Clarke, L.. PY - 1994. Y1 - 1994. N2 - Sexually dimorphic differences in voluntary sodium intake, locomotor activity, immobility in the swim test, and aggressive behavior were found to be altered in transgenic CD-1 mice that overexpressed transforming growth factor α (TGFα). In contrast to nontransgenic CD-1 mice, immobility in the swim test was longer and sodium intake higher in the male TGFα mice than in the female TGFα mice. These findings indicate that the male TGFα mice exhibited feminization of some behaviors. Furthermore, the male TGFα mice were highly aggressive. Castration reversed the behavioral effects in the adult male transgenic mice, but ovariectomy did not reverse the behavioral effects in the adult female transgenic mice. Thus the feminizing effect of TGFα on some nonreproductive ...
Targeting the expression of genes has emerged as a potentially viable therapeutic approach to human disease. In Alzheimers disease, therapies that silence the expression of tau could be a viable strategy to slow disease progression. We produced a novel strain of transgenic mice that could be used to assess the efficacy of gene knockdown therapies for human tau, in live mice. We designed a tetracycline-regulated transgene construct in which the cDNA for human tau was fused to ubiquitin and to luciferase to create a single fusion polyprotein, termed TUL. When expressed in brain, the TUL polyprotein was cleaved by ubiquitin-processing enzymes to release the luciferase as an independent protein, separating the half-life of luciferase from the long-lived tau protein. Treatment of bigenic tTA/TUL mice with doxycycline produced rapid declines in luciferase levels visualized by in vivo imaging and ex vivo enzyme measurement. This new mouse model can be used as a discovery tool in optimizing gene targeting
The Transgenic Mouse / ES Cell Shared Resource assists investigators in generating, maintaining and storing germline-altered mice. This resource, which has been in existence for over 23 years, has generated over 2700 transgenic founder mice from over 750 different DNA constructs and 5000 chimeric mice from over 800 different mouse ES cell clones. Prior to the discovery of the CRISPR/Cas system, the TMESCSR generated over 160 different gene-targeted mice. Since 2013, the TMESCSR has generated over 400 mutant pups by CRISPR/Cas injections from over 100 different projects. The Vanderbilt Transgenic Mouse/ES Cell Shared Resource (TMESCSR) provides services, consultation and collaborations to enable the generation, storage and regeneration of genetically altered mice at Vanderbilt. We have many years of experience in generating novel transgenic mouse models and are happy to discuss your project with you. The following procedures are currently provided on a fee-for-service basis: CRISPR/Cas9 Mouse
The Vanderbilt Genome Editing Resource (VGER) assists investigators in generating, maintaining and storing germline-altered mice in an efficient and cost-effective manner. The VGER provides services, consultation, and collaborations to enable the generation, storage and regeneration of genetically altered mice at Vanderbilt. This resource, which was previously called the Transgenic Mouse ESC Shared Resource (please see announcement letter), has been in existence for over 25 years and has generated 160 unique gene-targeted mice between 1993-2015 and more than 80 genome edited mice using CRISPR-Cas9 since 2014. We have many years of experience in generating novel transgenic mouse models and are happy to discuss your project with you. We provide the following services on a fee-for-service basis: CRISPR-Cas9 Mouse Editing Pronuclear Microinjection of DNAs Embryo Cryopreservation Sperm Cryopreservation In vitro Fertilization and Rederivation Genome-Editing Design and Analysis Services We are sorry, but we
Neural cell adhesion molecule-secreting transgenic mice display abnormalities in GABAergic interneurons and alterations in behavior.s profile, publications, research topics, and co-authors
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The biologic relevance of AICD to APP physiology or AD pathology has been proposed, but in vivo evidence that supports the hypothesis has been lacking. The current study was aimed at examining this hypothesis by expressing AICD postnatally and selectively in the forebrain and hippocampal regions of mouse brain. Here, we first show that AICD is present in brain membranes from normal control mice and can be detected by Western blot alone. We further demonstrate that the transgenic mice that were generated in this study express AICD at levels that are similar to those found in APP transgenic mice with FAD mutation, which are two- to threefold higher than in nontransgenic mice. These data strongly support the validity of our mouse model, which was verified further by observing increased expression of KAI1 gene in transgenic mice. Finally, we present evidence that the AICD transgenic mice display robust changes in GSK-3 signaling, which supports an in vivo role for AICD.. The finding that an ...
Missense mutations in APP and PS1 lead to familial forms of AD by different mechanisms. Most PS1 mutations shift γ‐secretase cleavage to increased Aβ42 production, which in turn accelerates cerebral amyloidosis in transgenic mice (Borchelt et al, 1997; Holcomb et al, 1998; Siman et al, 2000). An inverse correlation between the Aβ42 to Aβ40 ratio and the age of onset in familial AD has also been reported (Duering et al, 2005).. Mutations at position Leu 166 in PS1 lead to a severe course of AD pathology, with a very early onset in the third or fourth decade of life. So far, three mutations at position Leu 166 (L166A, L166P and L166H) have been described; the L166P mutation seems to be the most pathogenic (Ezquerra et al, 2000; Moehlmann et al, 2002; Pantieri et al, 2005). Expressing PS1‐L166P in transfected cells resulted in the highest Aβ42 to Aβ40 ratio among several PS1 mutations (Moehlmann et al, 2002). However, in contrast to other PS1 mutations, the L166P mutation has been shown ...
Here we report further our investigation of the role of BRE in HCC. HCC was chemically-induced in the transgenic (TTR-V5-BRE) and non-transgenic littermates, bred with C57BL/6, by intraperitoneal injection of diethylnitosamine (DEN) at 15 days postnatally. At 8 months after injection, the mice were sacrificed, and livers collected for determination of tumor number and maximal size, and for immunohistochemistry. Parts of each liver sample were also dissected visually into tumor and adjacent normal portions for Western Blot analysis of BRE expression. By comparison between the DEN-treated male transgenic mice (n=12) and non-transgenic littermate controls (n=8), we observed significantly increased tumor size shown by the former (p=0.049, Exact Wilcoxon Rank Sum test), with the median tumor size 2-fold larger than the latter. There was, however, no statistically significant difference between tumor numbers of the two groups. Female C57BL/6 mice are known to be less sensitive to DEN-treated ...
Tornavaca O, Pascual G, Barreiro ML, Grande MT, Carretero A, Riera M, Garcia-Arumi E, Bardaji B, González-Núñez M, Montero MA, López-Novoa JM, Meseguer A
Morphologic examinations of mammary neoplasias arising in BALB/c (H-2d) mice carrying the activated rat HER-2/neu oncogene (BALB-NeuT), and in FVB (H-2q) mice bearing the wild-type proto-oncogene (FVB-NeuN), indicate that both conditions result in a very human-like lobular carcinoma of alveolar type …
TY - JOUR. T1 - Immortalization of subpopulations of respiratory epithelial cells from transgenic mice bearing SV40 large T antigen. AU - Ikeda, K.. AU - Clark, J. C.. AU - Bachurski, C. J.. AU - Wikenheiser, K. A.. AU - Cuppoletti, J.. AU - Mohanti, S.. AU - Morris, R. E.. AU - Whitsett, J. A.. PY - 1994. Y1 - 1994. UR - http://www.scopus.com/inward/record.url?scp=0028041963&partnerID=8YFLogxK. UR - http://www.scopus.com/inward/citedby.url?scp=0028041963&partnerID=8YFLogxK. M3 - Article. AN - SCOPUS:0028041963. VL - 267. JO - American Journal of Physiology - Heart and Circulatory Physiology. JF - American Journal of Physiology - Heart and Circulatory Physiology. SN - 0363-6135. IS - 3 part 1. ER - ...
This interesting paper provides clear evidence that amyloid pathology in the double transgenic model causes axonopathy. The results suggest that intracellular Aβ accumulation in double transgenic mice may lead to trafficking defects in axons. While the results are compelling in the double transgenic, no such alterations are observed in single transgenic animals. Furthermore, amyloid pathology in spinal cord and axonopathy appear to be variable features that are not always present in AD patients. As the authors suggest, subtler alterations in signal transduction pathways, leading to misregulation of axonal transport and/or cytoskeletal disruption, may lead to motor deficits not only in AD, but also in other neurodegenerative conditions as well (Ebneth et al., 1998; Morfini et al., 2002; Pigino et al., 2003; Roy et al., 2005). Further studies will be required to determine if intracellular Aβ accumulation leads to motor dysfunction in AD.. ...
TY - JOUR. T1 - Recombinant ret oncogene products induce T lymphocyte proliferation, and suppress lymphoma derived from ret transgenic mice. AU - Yan, D.. AU - Isobe, K. I.. AU - Takahashi, M.. AU - Nakashima, I.. PY - 1994/1/1. Y1 - 1994/1/1. N2 - Recombinant ret oncogene products in complete Freunds adjuvant were injected into normal mice and ret oncogene transgenic mice. Spleen cells from the mice immunized with ret proteins were highly proliferated in vitro by the stimulation of ret proteins. The proliferating T cells have CD4+ phenotype and secreated CTLL-2 reactive interleukins. The peritoneal exudate cells from ret proteinimmunized mice suppressed ret-2 lymphoma, which was derived from ret lymphoma transgenic mice. The peritoneal exudate cells from ret melanoma transgenic mice did not suppress ret-2 lymphoma, when they were immunized with ret proteins.. AB - Recombinant ret oncogene products in complete Freunds adjuvant were injected into normal mice and ret oncogene transgenic mice. ...
Ke YD, van Hummel A, Stevens CH, Gladbach A, Ippati S, Bi M, Lee WS, Krüger S, van der Hoven J, Volkerling A, Bongers A, Halliday G, Haass NK, Kiernan M, Delerue F, Ittner LM. Short-term suppression of A315T mutant human TDP-43 expression improves functional deficits in a novel inducible transgenic mouse model of FTLD-TDP and ALS. Acta Neuropathologica 2015 Nov;130(5):661-78 PubMed 26437864 [https://dx.doi.org/10.1007/s00401-015-1486-0 ...
Background The mitogen-activated protein kinases, MAPKs for short, constitute cascades of signalling pathways involved in the regulation of several cellular processes that include cell proliferation, differentiation and motility. They also intervene in neurological processes like fear conditioning and memory. Since little remains known about the MAPK-Activated Protein Kinase, MAPKAPK5, we constructed the first MAPKAPK knockin mouse model, using a constitutive active variant of MAPKAPK5 and analyzed the resulting mice for changes in anxiety-related behaviour. Methods We performed primary SHIRPA observations during background breeding into the C57BL/6 background and assessed the behaviour of the background-bred animals on the elevated plus maze and in the light-dark test. Our results were analyzed using Chi-square tests and homo- and heteroscedatic T-tests; Results Female transgenic mice displayed increased amounts of head dips and open arm time on the maze, compared to littermate controls. In ...
Transgenic mice with cardiac-restricted overexpression of secretable TNF (MHCsTNF) develop progressive LV wall thinning and dilation accompanied by an increase in cardiomyocyte apoptosis and a progressive loss of cytoprotective Bcl-2. To test whether cardiac-restricted overexpression of Bcl-2 would prevent adverse cardiac remodeling, we crossed MHCsTNF mice with transgenic mice harboring cardiac-restricted overexpression of Bcl-2. Sustained TNF signaling resulted in activation of the intrinsic cell death pathway, leading to increased cytosolic levels of cytochrome c, Smac/Diablo and Omi/HtrA2, and activation of caspases -3 and -9. Cardiac-restricted overexpression of Bcl-2 blunted activation of the intrinsic pathway and prevented LV wall thinning; however, Bcl-2 only partially attenuated cardiomyocyte apoptosis. Subsequent studies showed that c-FLIP was degraded, that caspase-8 was activated, and that Bid was cleaved to t-Bid, suggesting that the extrinsic pathway was activated concurrently in ...
Full Text - CYLD is a deubiquitinating enzyme known for its role as a tumor suppressor whose mutation leads to skin appendages tumors and other cancers. In this manuscript we report that the tumor suppressor CYLD, similarly to other renowned tumor suppressor genes, protects from premature aging and cancer. We have generated transgenic mice expressing the mutant CYLDC/S protein, lacking its deubiquitinase function, under the control of the keratin 5 promoter, the K5-CYLDC/S mice. These mice express the transgene in different organs, including those considered to be more susceptible to aging, such as skin and thymus. Our results show that K5-CYLDC/S mice exhibit epidermal, hair follicle, and sebaceous gland alterations; and, importantly, they show signs of premature aging from an early age. Typically, 3-month-old K5-CYLDC/S mice exhibit a phenotype characterized by alopecia and kyphosis, and, the histological examination reveals that transgenic mice show signs
Nuclear factor κ-light chain enhancer of activated B cells (NF-κB) is a transcription factor that can be activated through canonical or non-canonical signaling pathways, and activation of both pathways has been observed in lung adenocarcinoma tumors. However, the mechanistic links between canonical and non-canonical NF-κB signaling and lung tumorigenesis have not been fully elucidated. Using transgenic mouse models, we demonstrate that canonical NF-κB signaling promotes epidermal growth factor receptor (EGFR)-mediated tumor formation through paracrine signaling to the inflammatory microenvironment, with depletion studies identifying macrophages as a critical cell type promoting EGFR-driven lung tumorigenesis. To study non-canonical NF-κB signaling, we developed a novel transgenic mouse model with inducible over-expression of the non-canonical NF-κB component p52 in the airway epithelium. After injection with the lung carcinogen urethane, p52 over-expression led to increased tumor number, ...
Cystic fibrosis is characterized by dehydration of the airway surface liquid layer with persistent mucus obstruction. |i|Th2|/i| immune responses are often manifested as increased mucous cell density (mucous cell metaplasia) associated with mucus obstruction. IL-33 is a known inducer of |i|Th2|/i| i …
Increased specific activity of Tg-HDL in the presence of human apoA-I and Hpr. (A) Western blot with serial dilutions of plasma from HuapoA-I mice expressing Hpr and apoL-I from a single plasmid (HuapoA-I:Hpr:apoL-I) and normal human plasma. Hpr and apoL-I were detected with monoclonal antibodies. (B) Survival kinetics of naive mice infected with T. b. brucei ILTat 1.25 and then given 300 μl plasma i.v. from HuapoA-I mice expressing Hpr (black diamond; n = 3), apoL-I (black square; n = 3), or both Hpr and apoL-I from a single plasmid (black circle, Hpr:apoL-I; n = 3). The protection obtained by normal human plasma (dilution 1/8) is indicated by the inverted triangle. (C) A280 profile of KBr-purified lipoproteins from human (dashed line) and HuapoA-I mouse plasma expressing Hpr:apoL-I (red line), Hpr (green line), and apoL-I (blue line) separated by size on a Superdex 200 column; fractions 6-7 are void, fractions 8-9 are human LDL, fractions 10-14 are HDLs, and fraction 15 is albumin. (D) ...
The present study showed that levels of BMP6 were increased approximately twofold to fourfold in the hippocampus of patients with AD and in APP tg mice compared to controls; however, no significant differences were detected in the mRNA levels of two other BMPs, BMP2 and BMP7. A striking pattern of BMP6 distribution was also observed in plaque-containing regions of the hippocampus in both AD patients and APP tg mice, where Aβ-containing plaques were surrounded by a ring-like pattern of BMP6 immunoreactivity. Since BMP6 is a secreted protein, and its primary reported role in the brain is in regulating developmental neurogenesis, it is possible that abnormally elevated levels of this protein in AD might affect adult neurogenesis in the hippocampus.. It is important to note that neurogenesis persists in the aged brain; however, its rate declines with increasing age, as revealed by previous studies in rodents (Kuhn et al., 1996; Kempermann et al., 1998), nonhuman primates (Gould et al., 1999), and ...
AP-2 transcription factors play pivotal roles in orchestrating embryonic development by influencing the differentiation, proliferation, and survival of cells. Furthermore, AP-2 transcription factors have been implicated in carcinogenesis, a process where the normal growth and differentiation program of cells is disturbed. To experimentally address the potential involvement of AP-2 in mammary gland tumorigenesis, we generated mice overexpressing AP-2gamma by transgenesis using the mouse mammary tumor virus-long terminal repeat as the transgene-driving promoter unit. In the mammary gland, transgene expression elicited a hyperproliferation that, however, was counterbalanced by the enhanced apoptosis of epithelial cells leading to a hypoplasia of the alveolar epithelium during late pregnancy. In addition, secretory differentiation was impaired, resulting in a lactation failure. In male transgenic mice, the seminal vesicles were sites of strong transgene expression. There the effects of AP-2gamma on ...
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We previously found that adult Kir6.2G132S transgenic female mice (at 8 weeks of age) exhibit hyperglycemia with hypoinsulinemia caused by accelerated apoptosis (26). In the current study, we found that hyperglycemia with hypoinsulinemia in Kir6.2G132S transgenic mice recovers significantly with age. We also show that pancreatic insulin content and the number of β-cells both increase as the Kir6.2G132S transgenic mice become older. The presence of the Kir6.2G132S mutation in the older Kir6.2G132S transgenic mice excludes the possibility that loss of the transgene is responsible for the recovery of β-cell number in the older Kir6.2G132S transgenic mice. In fact, the insulin response to glucose loading was absent in the older Kir6.2G132S transgenic mice. Hence, the recovery of blood glucose and serum insulin are most likely caused by the restoration of pancreatic insulin content resulting from the increased β-cell number in the older Kir6.2G132S transgenic mice. In a preliminary study, we found ...
ACE, as well as all other components of the renin-angiotensin system (RAS), is expressed in a variety of organs, including the kidney (16,19,20). The generation of Ang II by the tissue RAS suggests a role of the peptide as a paracrine modulator of organ function in addition to its function as a circulating hormone. Distinguishing between the importance of systemic and locally produced Ang II is not trivial because pharmacologic inhibitors as well as knockout approaches affect both modes of angiotensin action. Our study used transgenic mice that had been genetically engineered to express a single ACE isoform in a cell type-specific location on the background of an ACE null genotype (10,13). This approach seems well suited to assess the notion of a location-specific role of ACE and to determine the contribution of this restricted expression pattern to the overall health and organ function of an animal.. Previous studies in AT1A and ACE knockout mice as well as earlier experiments with ACE ...
We have used transgenic mouse technology to establish immortalized hepatoma cell lines stably secreting heterologous proteins, such as human α1-antitrypsin and human factor IX. Hepatocyte-specific...
Dr. Yans research focuses on investigating the cellular and molecular mechanisms of cellular stress and survival in neurodegenerative disorders relevant to Alzheimers disease (AD) and Parkinson disease. She has first identified the specific cellular targets (RAGE, receptor for advanced glycation end product; and ABAD, amyloid binding alcohol dehydrogenase) of amyloid-beta peptide (Aβ) and found the evidence of Aβ-mediated neuronal stress. She developed a novel transgenic mouse model relevant to AD and tested the role of RAGE and ABAD in Aβ-mediated cellular perturbation in those AD type mouse models. She was the first to describe the RAGE and ABAD as the functional binding proteins for Aβ. Dr. Yan and her research team are the major group investigating these paradigms. Dr. Yan and her research team have provided evidence that cell surface molecule (RAGE) and mitochondrial enzyme (ABAD) serve as cofactors for promoting and exaggerating neuronal and mitochondrial toxicity in an Aβ-rich ...
In 2007, McCray, Jr. et al from the University of Iowa published a study in which they introduced a vector carrying a human ACE2-coding sequence into wild-type mice and subsequently developed a successful hACE2 transgenic mouse strain. ACE2 expression, which is regulated by the human cytokeratin 18 (K18) promoter in epithelial cells, was observed in the initially infected airway epithelial cells. Studies showed that the K18-hACE2 transgenic mouse infected with a human SARS-CoV strain via intranasal inoculation would not survive. The infection would begin in the airway epithelium, spread to the alveoli and finally out of the lungs to the brain. The infection causes infiltration of macrophages and lymphocytes in the lungs and up-regulation of pro-inflammatory cytokines and chemokines in the lungs and brain. Three to five days following infection, K18-hACE2 mice began to lose weight and become lethargic with labored breathing. All died within seven days. These observations support that transgene ...
Numerous studies using rats and mice have been conducted to examine the cellular mechanisms and processes involved in Alzheimers disease.. A study conducted by Um et al.[105] used transgenic mice Tg-NSE/hPS2m, which expressed the human PS2 mutation and compared them to control mice, who were labeled as non-Tg. Mice were sacrificed from each group and brains were removed and separated so the hippocampus and the extracted mitochondria could be analyzed using western blot analysis.[105] Tunel staining was also used to detect apoptosis.[105] It was found that phosphorylation levels of tau at the Ser404, Ser202, and Thr231 residues in the hippocampus in Tg mice were enhanced.[105] Increased phosphorylation levels of JNK1/2 and p38MAPK along with decreased levels of ERK1/2 phosphorylation were found in transgenic mice (Tg).[105] Tg mice also had higher levels of COX-2 proteins and higher levels of caspase-3 protein levels than control mice.[105] Cytochrome C and Bax protein levels were higher and ...
Nikolaos Svoronos is the author of this article in the Journal of Visualized Experiments: Initiation of Metastatic Breast Carcinoma by Targeting of the Ductal Epithelium with Adenovirus-Cre: A Novel Transgenic Mouse Model of Breast Cancer
Tom L. Stephen is the author of this article in the Journal of Visualized Experiments: Initiation of Metastatic Breast Carcinoma by Targeting of the Ductal Epithelium with Adenovirus-Cre: A Novel Transgenic Mouse Model of Breast Cancer
46 A novel transgenic mice has been generated which overexpresses the constitutively activated mutant form of human Rac 1 (Rac-CA), a small GTP-binding protein of the Rho family, in smooth muscle cells. Experiments were conducted to determine whether chronic activation of Rac-1 affects blood pressure (BP) and heart rate (HR). Male Rac-CA and control mice were prepared for chronic cardiovascular monitoring using carotid arterial catheters infused with heparinized saline. BP and HR were measured continuously for more than 4 days. Results show an increased BP in Rac-CA with no change in HR or water intake. BP was consistently higher in Rac-CA mice (Figure), averaging 122±4 vs 109±4 mm Hg during the dark phase (Rac-CA vs Control). HR showed a night/day rhythm with no differences between the Rac-CA and Controls, respectively, 640±15 vs 610±11 bpm (dark) and 572 ±39 vs 578 ±18 (light). These results illustrate that alteration in intracellular signaling (reactive oxygen species) in vascular ...
The risk of heart failure following myocardial infarction is higher in diabetic patients than nondiabetic patients. The mammalian target of rapamycin (mTOR), a key downstream molecule of insulin-phosphoinositide 3-kinase (PI3K)-Akt signaling pathway, plays an important role in cardioprotection. However, the role of cardiac mTOR in ischemic injury in metabolic syndrome has not been well defined. To address this question, we studied the effect of overexpressing cardiac mTOR on cardiac function following ischemia/reperfusion (I/R) in mice with high-fat diet (HFD)-induced obesity. In this study, we used transgenic mice with cardiac-specific overexpression of mTOR (mTOR-Tg) as reported previously. mTOR-Tg and WT mice at 6 weeks old were fed HFD (60% fat by calories) ad libitum for 14 weeks. Control mTOR-Tg and WT mice were fed a normal chow diet (NCD). At 14 weeks after HFD, glucose and insulin tolerance tests demonstrated that HFD generated glucose intolerance and insulin resistance in both mTOR-Tg ...
The signals that determine the size and duration of the primary T cell immune response are not well defined. We studied CD4 T cells at an important checkpoint in their development: when they have become effectors and are ready to rapidly mediate effector functions, both via direct interaction with antigen (Ag)-presenting cells and via cytokine production. We determined the effects of specific Ag and the cytokines interleukin (IL) 2 and transforming growth factor (TGF) beta 1 on T helper cell type 2 (Th2) effector apoptosis versus expansion. Th2-polarized effector cells were generated in vitro from naive CD4 T of T cell receptor transgenic mice, and then restimulated with or without peptide Ag plus Ag-presenting cells and cytokines. In the absence of added cytokines, effector cells cultured without Ag died of apoptosis after 4-7 d. Paradoxically, Ag both induced proliferation and high levels of cytokine synthesis and accelerated effector cell death. IL-2 directly induced proliferation of ...
Absence of lung fibrosis in TGFβ1 overexpressing mice. Figure A shows haematoxylin and eosin staining of Tr+ TGFβ1 transgenic lung tissue. Of note was the abs
In this study, we presented a new line of α-syn A53T conditional transgenic mice to specifically investigate the underlying subcellular and molecular pathogenic pathways leading to α-syn-mediated dysfunction and degeneration of mDA neurons. These A53T mice developed profound movement impairments, especially the rearing activities, perhaps reflecting a severe dysfunction of the nigrostriatal dopaminergic system. Indeed, substantial reduction of dopamine release was observed in the dorsal striatum of 1-month-old A53T mice. Moreover, robust and progressive mDA neurodegeneration was apparent in the 6-month-old mutant mice. Perhaps more interestingly, we identified Nurr1 as an important downstream molecular target of α-syn, in which α-syn promoted a proteasome-dependent degradation of Nurr1 protein, resulting in a preferential dysfunction and loss of mDA neurons. Conversely, a modest suppression of proteasome activities in the mDA neurons ameliorated the α-syn-induced Nurr1 degradation and mDA ...
Principal Investigator:NISHIMURA Yasuharu, Project Period (FY):1989 - 1991, Research Category:Grant-in-Aid for Developmental Scientific Research., Research Field:Immunology
TY - JOUR. T1 - Bone marrow-derived stem cells and hepatocarcinogenesis in hepatitis B virus transgenic mice. AU - Barone, Michele. AU - Scavo, Maria Principia. AU - Maiorano, Eugenio. AU - Di Leo, Alfredo. AU - Francavilla, Antonio. PY - 2014/3. Y1 - 2014/3. N2 - Background: Several studies have demonstrated that cancer can develop with the contribution of bone marrow-derived cancer stem cells. We evaluated the possible involvement of bone marrow-derived stem cells in hepatocarcinogenesis in a hepatitis B virus (HBV) transgenic mouse model. Methods: Bone marrow cells from wild type male mice were transplanted into sublethally irradiated, female, HBV transgenic mice with hepatocarcinoma nodules. Four months later, liver tissue was examined to localize neoplastic nodules/foci and characterize cells by evaluating the Y-chromosome and the hepatocyte lineage marker hepatocyte nuclear factor-1 (HNF1), as well as the HBsAg encoding gene (HBs-Eg) and HBsAg protein (HBs-Pr) (present only in cells of ...
Injury to epidermis and other stratified epithelia triggers profound but transient changes in the pattern of keratin expression. In postmitotic cells located at the wound edge, a strong induction of K6, K16, and K17 synthesis occurs at the expense of the keratins produced under the normal situation. The functional significance of these alterations in keratin expression is not known. Here, we report that overexpression of a wild-type human K16 gene in a tissue-specific fashion in transgenic mice causes aberrant keratinization of the hair follicle outer root sheath and proximal epidermis, and it leads to hyperproliferation and increased thickness of the living layers (acanthosis), as well as cornified layers (hyperkeratosis). The pathogenesis of lesions in transgenic mouse skin begins with a reorganization of keratin filaments in postmitotic keratinocytes, and it progresses in a transgene level-dependent fashion to include disruption of keratinocyte cytoarchitecture and structural alterations in ...
Handling of Mice. The Tg2576 mice were developed by Karen Hsaio (Hsiao et al., 1996) and licensed from the Mayo Foundation for Medical Education and Research (Rochester, MN). Male Tg2576 transgenic mice were bred to normal C57BL6/SJL females at the Bristol-Myers Squibb facility in Wallingford, CT. Mice were housed with a 6:00 AM to 6:00 PM light/dark cycle and allowed free access to food and water. Both male and female mice were used in these studies, and although no differences in Aβ were observed between them, only one sex was used in a single study. Young Tg2576 mice were used between 3 and 6 months of age, whereas aged animals were used at 14 to 17 months. BMS-299897 was synthesized by the Medicinal Chemistry groups of SIBIA Neurosciences, Inc. (now Merck Research Laboratories, San Diego, CA) and Bristol-Myers Squibb. Animals were dosed by oral gavage in a volume of 6 ml/kg in polyethylene glycol, average molecular weight of 400, or a vehicle consisting of 10% propylene glycol, 7.5% ...
106 Gallbladder carcinomas carry poor prognosis and difficulties with treatment, often due to late stage diagnosis, highly malignant nature, and limited knowledge regarding the pathogenesis. Overexpression of erbB2 in gallbladder epithelial cells in BK5.erbB2 transgenic mice leads to development of adenocarcinoma in 90% of the homozygous transgenic mice. This transgenic mouse model provided a useful tool for investigating the mechanism of erbB2 induced development of gallbladder carcinoma. We detected that erbB2 overexpression in transgenic gallbladder epithelial cells was associated with a high level of EGFR and both erbB2 and EGFR were constitutively activated. We further analyzed the downstream of erbB2/EGFR signaling in gallbladder epithelial cells from BK5.erbB2 mice to investigate the mechanism of carcinogenesis. Immunohistochemical analysis revealed that activated Akt was predominantly nuclear in gallbladder epithelial cells from the transgenic mice, but not in nontransgenic mice. Western ...
TY - JOUR. T1 - Attenuation of length dependence of calcium activation in myofilaments of transgenic mouse hearts expressing slow skeletal troponin I. AU - Arteaga, Grace M.. AU - Palmiter, Kimberly A.. AU - Leiden, Jeffrey M.. AU - Solaro, R. John. PY - 2000/1/1. Y1 - 2000/1/1. N2 - 1. We compared sarcomere length (SL) dependence of the Ca2+-force relation of detergent-extracted bundles of fibres dissected from the left ventricle of wild-type (WT) and transgenic mouse hearts expressing slow skeletal troponin I (ssTnI-TG). Fibre bundles from the hearts of the ssTnI-TG demonstrated a complete replacement of the cardiac troponin I (cTnI) by ssTnI. 2. Compared to WT controls, ssTnI-TG fibre bundles were more sensitive to Ca2+ at both short SL (1· ± 0·1 μm) and long SL 2·3 ± 0·1 μm). However, compared to WT controls, the increase in Ca2+ sensitivity (change in half-maximally activating free Ca2+; ΔEC50) associated with the increase in SL was significantly blunted in the ssTnI-TG ...
High fat/high cholesterol diets exacerbate β-amyloidosis in mouse models of Alzheimers disease (AD). It has been impossible, however, to study the relationship between atherosclerosis and β-amyloidosis; in those models because such mice were on atherosclerosis-resistant genetic backgrounds. Here we report the establishment of AD model mice, B6Tg2576, that are prone to atherosclerosis. B6Tg2576 mice were produced by back-crossing Tg2576 mice, an AD mouse model overexpressing human amyloid β-protein precursor with the Swedish double mutation, to C57BL/6 mice, a strain susceptible to diet-induced atherosclerosis. An atherogenic diet induced aortic atherosclerosis and exacerbated cerebral β-amyloidosis in B6Tg2576 mice. Compared with age-matched non-transgenic littermates, B6Tg2576 mice developed significantly more diet-induced aortic atherosclerosis. Unexpectedly, normal diet-fed B6Tg2576 mice also developed fatty streak lesions (early atherosclerosis) in the aorta. The aortic atherosclerotic ...
Marxreiter, Franz; Ettle, Benjamin; May, Verena E. L.; Esmer, Hakan; Patrick, Christina; Kragh, Christine Lund; Klucken, Jochen; Winner, Beate; Riess, Olaf; Winkler, Juergen; Masliah, Eliezer; Nuber, Silke ...
TY - JOUR. T1 - HLA Class II Transgenic Mice Mimic Human Inflammatory Diseases. AU - Mangalam, Ashutosh K.. AU - Rajagopalan, Govindarajan. AU - Taneja, Veena D. AU - David, Chella S.. PY - 2008. Y1 - 2008. N2 - Population studies have shown that among all the genetic factors linked with autoimmune disease development, MHC class II genes on chromosome 6 accounts for majority of familial clustering in the common autoimmune diseases. Despite the highly polymorphic nature of HLA class II genes, majority of autoimmune diseases are linked to a limited set of class II-DR or -DQ alleles. Thus a more detailed study of these HLA-DR and -DQ alleles were needed to understand their role in genetic predisposition and pathogenesis of autoimmune diseases. Although in vitro studies using class-II restricted CD4 T cells and purified class II molecules have helped us in understanding some aspects of HLA class-II association with disease, it is difficult to study the role of class II genes in vivo because of ...
A truncated human O6-alkylguanine-DNA-alkyltransferase (ATase) cDNA was ligated into an expression vector under the control of the mouse metallothionein-1 gene promotor and upstream of part of the human growth hormone gene to provide splice and polyadenylation signals. Transfection of this construct into human cells resulted in very high levels of ATase expression (more than 300 fmoles/mg protein versus less than 2 fm/mg protein in parent vector transfected control cells). Microinjection of a 4.2 kb fragment of this vector into B6D2F2 mouse embryos and implantation of survivors into pseudopregnant females has so far generated 35 offspring. Southern analysis of tail tip DNA has shown that 11 of the offspring are transgenic for the human ATase gene, between 1 and at least 30 copies of the gene being detected. Human ATase transcripts were detected in total RNA extracted from liver obtained from two male transgenic mice by partial hepatectomy. Cell free extracts of liver samples from five transgenic ...
Introduction: We have previously used a novel transgenic mouse model that expresses an inducible dominant negative mutation of the TGF-β type II receptor (DnTGFβRII) to demonstrate that blocking pro-fibrogenic TGF-β signaling reduces pressure overload-induced interstitial collagen deposition in the heart. The current study utilized DnTGFβRII mice to test the hypothesis that collagen deposition in the pressure overloaded heart is required to maintain structure and prevent cardiac dilation and dysfunction.. Methods: 8 -10 wk old male DnTGFβBRII mice and nontransgenic control (NTG) mice were given 25 mM ZnSO4 in drinking water to induce the expression of DnTGFβRII gene 1 wk prior to transverse aortic constriction (TAC). 120 days after TAC or sham operation, left ventricular (LV) mass, dimension and function were assessed by echocardiography using a high frequency ultrasound probe and interstitial collagen content was assessed in picrosirius red stained sections of LV by light microscopy with ...
We previously described an enhancer variant of Moloney murine leukaemia virus (M-MuLV), ΔMo + SV M-MuLV, in which the enhancers of MuLV have been deleted and replaced with the enhancers of the simian virus 40 (SV40). When this virus is injected into neonatal NIH Swiss mice, pre-B and B-lymphoblastic lymphomas develop with a latency of 17 months. Van Lohuizen et al. (1989) described a line of transgenic mice that carry an activated pim-1 proto-oncogene transgene (Eµ pim-1). They also reported that Eµ pim-1 transgenic mice show greatly accelerated lymphoma development when infected with wild-type M-MuLV at birth. In these experiments, neonatal Eµ pim-1 transgenic mice were infected intraperitoneally with ΔMo + SV M-MuLV. Marked acceleration of T-lymphoid leukaemia was seen. However, 10 of the 11 tumours analysed were found to be negative for the SV40 enhancers, but they still contained M-MuLV DNA as measured by Southern blot analysis. The LTRs on viruses cloned from two such tumours (as well as on
The recruitment of monocytes and their differentiation into macrophages at sites of inflammation are key events in determining the outcome of the inflammatory response and initiating the return to tissue homeostasis. To study monocyte trafficking and macrophage differentiation in vivo, we have generated a novel transgenic reporter mouse expressing a green fluorescent protein (GFP) under the control of the human CD68 promoter. CD68-GFP mice express high levels of GFP in both monocyte and embryo-derived tissue resident macrophages in adult animals. The human CD68 promoter drives GFP expression in all CD115(+) monocytes of adult blood, spleen, and bone marrow; we took advantage of this to directly compare the trafficking of bone marrow-derived CD68-GFP monocytes to that of CX3CR1(GFP) monocytes in vivo using a sterile zymosan peritonitis model. Unlike CX3CR1(GFP) monocytes, which downregulate GFP expression on differentiation into macrophages in this model, CD68-GFP monocytes retain high-level GFP
en] Using transgenic mice constitutively expressing the human inducible Hsp70, we examined the role of Hsp70 on cell survival after focal cerebral ischemia. Twenty-four hours after permanent occlusion of the middle cerebral artery, no difference in infarct area was detected between Hsp70-transgenic and non-transgenic mice. In the non-transgenic mice, many pyramidal neurons of the ipsilateral hippocampus were observed to be pyknotic. However, in all Hsp70-transgenic mice, hippocampal pyramidal neurons showed normal morphology and no evidence of pyknosis. This suggests that constitutive expression of Hsp70 reduces the extent of damage following permanent middle cerebral artery occlusion ...
Nox2-containing NADPH oxidases are reported to be involved in the development of cardiac fibrosis in response to chronic angiotensin II infusion, but the cellular source(s) of Nox2 involved in fibrosis remains unclear. We investigated the role of endothelial Nox2 in angiotensin II-induced left ventricular hypertrophy (LVH). Male transgenic mice with endothelial-specific overexpression of Nox2 were compared with matched wild-type (wt) littermates after angiotensin II (1.1 mg/kg per day) or saline infusion for 14 days. Basal blood pressure and left ventricular NADPH oxidase activity were similar in wt and transgenic mice. After angiotensin II infusion, both wt and transgenic groups developed similar hypertension (170.2±11.6 vs 170.4±12.3 mm Hg; n=10) and hypertrophy (left ventricular/body weight ratio 4.8±0.2 vs 4.7±0.2 mg/g; and echocardiographic septal thickness increased by 34% wt and 37% transgenic mice; n,10). NADPH oxidase activity was higher in angiotensin II-infused transgenic compared ...
Transgenic mice were produced that carried in their germlines rearranged kappa and/or mu genes with V kappa and VH regions from the myeloma MOPC-167 kappa and H genes, which encode anti-PC antibody. The mu genes contain either a complete gene, including the membrane terminus (mu genes), or genes in which this terminus is deleted and only the secreted terminus remains (mu delta mem genes). The mu gene without membrane terminus is expressed at as high a level as the mu gene with the complete 3 end, suggesting that this terminus is not required for chromatin activation of the mu locus or for stability of the mRNA. The transgenes are expressed only in lymphoid organs. In contrast to our previous studies with MOPC-21 kappa transgenic mice, the mu transgene is transcribed in T lymphocytes as well as B lymphocytes. Thymocytes from mu and kappa mu transgenic mice display elevated levels of M-167 mu RNA and do not show elevated levels of kappa RNA, even though higher than normal levels of M-167 kappa ...
Tissue-specific gene inactivation using the Cre-loxP system has become an important tool to unravel functions of genes when the conventional null mutation is lethal. We report here the generation of a transgenic mouse line expressing Cre recombinase in endothelial cells. In order to avoid the production and screening of multiple transgenic lines we used embryonic stem cell and embryoid body technology to identify recombinant embryonic stem cell clones with high, endothelial-specific Cre activity. One embryonic stem cell clone that showed high Cre activity in endothelial cells was used to generate germline chimeras. The in vivo efficiency and specificity of the transgenic Cre was analysed by intercrossing the tie-1-Cre line with the ROSA26R reporter mice. At initial stages of vascular formation (E8-9), LacZ staining was detected in almost all cells of the forming vasculature. Between E10 and birth, LacZ activity was detected in most endothelial cells within the embryo and of extra-embryonic ...
Background: Degeneration of the locus coeruleus (LC), the major noradrenergic nucleus in the brain, occurs early and is ubiquitous in Alzheimers disease (AD). Experimental lesions to the LC exacerbate AD-like neuropathology and cognitive deficits in several transgenic mouse models of AD. Because the LC contains multiple neuromodulators known to affect amyloid β toxicity and cognitive function, the specific role of noradrenaline (NA) in AD is not well understood. Methods: To determine the consequences of selective NA deficiency in an AD mouse model, we crossed dopamine β-hydroxylase (DBH) knockout mice with amyloid precursor protein (APP)/presenilin-1 (PS1) mice overexpressing mutant APP and PS1. Dopamine β-hydroxylase (-/-) mice are unable to synthesize NA but otherwise have normal LC neurons and co-transmitters. Spatial memory, hippocampal long-term potentiation, and synaptic protein levels were assessed. Results: The modest impairments in spatial memory and hippocampal long-term ...
We hypothesized that AT2R counteracts the growth-promoting effect of Ang II mediated by AT1R via apoptosis in the myocardium. To evaluate the effects of emphasized AT2R stimulation in vivo, we used transgenic mice overexpressing AT2R in a cardiac-specific manner. We evaluated both the sole Ang II effects (subpressor dose of Ang II) and the effects of Ang II and hemodynamic overload mediated by Ang II (pressor dose of Ang II) on cardiomyocyte apoptosis. Furthermore, we used L158809, a specific AT1R antagonist, to eliminate the effects through AT1R. This AT1R antagonist also causes upregulation of endogenous Ang II,27 which selectively stimulates overexpressed AT2R. Therefore, in this experimental system, we could stimulate AT2R selectively and maximally. In none of the conditions, however, did the number of TUNEL-positive nuclei in TG mice differ from that in WT mice, indicating that Ang II infusion for 28 days did not induce apoptosis in the mouse heart. At least, considering that we probably ...
The cAMP-response element-binding protein (CREB) is activated by phosphorylation on serine 133 and mediates the proliferative response to a number of different signals. A mutant CREB with a serine to alanine substitution at position 133 (CREBM1) functions as a dominant-negative inhibitor. Transgenic mice that express the dominant-negative CREB protein in B lymphocytes were developed as a means to study the effects of the inhibition of CREB function on B-cell proliferation and survival. We have shown previously that CREB up-regulates Bcl-2 expression in B cells in response to activation signals. B cells from CREBM1 transgenic mice expressed lower levels of Bcl-2 with and without stimulation. Proliferation of B cells from the transgenic mice was impaired in part by lack of induction of activator protein 1 (AP1) transcription factors. B cells from the transgenic mice were more susceptible to induction of apoptosis with several different agents, consistent with the decreased expression of Bcl-2. ...
The ubiquitin-proteasome system degrades most intracellular proteins, including misfolded proteins. Proteasome functional insufficiency (PFI) was observed in experimental proteinopathies and implicated in many human common diseases but its pathogenic role has not been established because a measure to enhance proteasome function in the cell has not been reported until very recently. We have recently discovered that overexpression of proteasome activator 28α (PA28α) enhances proteasome-mediated removal of abnormal proteins in the cell and protects against oxidative stress in cultured cardiomyocytes (FASEB J 2011; 25(3):883-93). Here we have extended the in vitro discoveries to intact animals. First, we created inducible transgenic mice with cardiomyocyte-restricted PA28α overexpression (CR-PA28αOE). CR-PA28αOE does not alter the homeostasis of normal proteins and cardiac function but increases the degradation of a surrogate misfolded protein in the heart. This marks the establishment of the ...
Using a transgenic mouse model in which the human B-myb cDNA was driven by the basal CMV promoter, this study demonstrates for the first time that B-Myb leads to markedly reduced neointima formation after mechanical injury to the vasculature and to decreased α1(I) collagen mRNA expression in the aorta and femoral artery of adult animals. Three independent transgenic mouse lines were generated, all of which apparently developed and bred normally. An inverse relationship between levels of B-Myb protein and expression of α1(I) collagen mRNA in the adult aorta was demonstrated. A decrease was also seen in α2(V) collagen mRNA levels in the aorta and in cultured aortic SMCs isolated from adult transgenic B-myb mice (data not shown). Importantly, when a femoral artery model of endothelial denudation was used, a dramatic reduction in neointima formation was observed in arteries of transgenic versus WT mice 4 weeks after injury. After injury, the neointimal area and the ratio of the areas of the ...
During a T cell-dependent Ab response, B cells undergo Ab class switching and V region hypermutation, with the latter process potentially rendering previously innocuous B cells autoreactive. Class switching and hypermutation are temporally and anatomically linked with both processes dependent on the enzyme, activation-induced deaminase, and occurring principally, but not exclusively, in germinal centers. To understand tolerance regulation at this stage, we generated a new transgenic mouse model expressing a membrane-tethered gamma2a-reactive superantigen (gamma2a-macroself Ag) and assessed the fate of emerging IgG2a-expressing B cells that have, following class switch, acquired self-reactivity of the Ag receptor to the macroself-Ag. In normal mice, self-reactive IgG2a-switched B cells were deleted, leading to the selective absence of IgG2a memory responses. These findings identify a novel negative selection mechanism for deleting mature B cells that acquire reactivity to self-Ag. This process ...
Alzheimers Imaging Consortium IC-P: Poster Presentations Background: Rosiglitazone, a peroxisome proliferator-activated receptor copy. Because of their high iron content, plaques typically appear as hypo- [gamma] (PPAR[gamma]) agonist, has an anti-inflammatory effect in the intense spots on T2-weighted scans. One of the challenges in imaging brain, decreasing interleukin-1[beta] concentrations in hippocampus and re- plaques is to achieve high-enough resolution and contrast to detect these storing the age-related deficit in long-term potentiation. It also attenuates 50-mm large lesions. While most high-field systems can reach high resolu- learning and memory deficits in a mouse model of Alzheimers disease. Ev- tion, the lack of contrast between the plaques and the parenchyma often idence suggests that activation of microglia and astrocytes contribute to age- impedes their detection. Methods: Transgenic mice over-expressing muta- related neuroinflammatory changes. In this study, relaxometry ...
TY - JOUR. T1 - Dlx5/6-Enhancer Directed Expression of Cre Recombinase in the Pharyngeal Arches and Brain. AU - Ruest, Louis Bruno. AU - Hammer, Robert E. AU - Yanagisawa, Masashi. AU - Clouthier, David E.. PY - 2003/12. Y1 - 2003/12. N2 - Dlx5 and Dlx6, two members of the Distalless gene family, are required for development of numerous tissues during embryogenesis, including facial and limb development. This gene pair is expressed in tandem, transcribed toward each other and separated by a short intergenic region containing multiple putative enhancers. Targeted inactivation of Dlx5 and Dlx6 in mice results in multiple developmental defects in craniofacial and limb structures, suggesting that these genes are crucial for aspects of both neural crest and nonneural crest development. To further investigate potential developmental roles of Dlx5 and Dlx6, we used one of the Dlx5/6 intergenic enhancers to drive Cre recombinase expression in transgenic mice. Crossing Dlx5/6-Cre transgenic mice with ...
The present study assessed the potential functions of interleukin (IL)-32α on inflammatory arthritis and endotoxin shock models using IL-32α transgenic (Tg) mice. The potential signaling pathway for the IL-32-tumor necrosis factor (TNF)α axis was analyzed in vitro. IL-32α Tg mice were generated under control of a ubiquitous promoter. Two disease models were used to examine in vivo effects of overexpressed IL-32α: Toll-like receptor (TLR) ligand-induced arthritis developed using a single injection of lipopolysaccharide (LPS) or zymosan into the knee joints; and endotoxin shock induced with intraperitoneal injection of LPS and D-galactosamine. TNFα antagonist etanercept was administered simultaneously with LPS in some mice. Using RAW264.7 cells, in vitro effects of exogenous IL-32α on TNFα, IL-6 or macrophage inflammatory protein 2 (MIP-2) production were assessed with or without inhibitors for nuclear factor kappa B (NFκB) or mitogen-activated protein kinase (MAPK). Single injection of LPS, but
We have used an aggrecan gene enhancer to generate a transgenic murine line (Acan-CreER-Ires-Luc) expressing firefly luciferase and tamoxifen activatable Cre recombinase (Cre-ER(T2) ). The expression and efficiency of the inducible Cre recombinase activity were tested in double transgenic mice created by crossing the Acan-CreER-Ires-Luc line with a Rosa26-lacZ reporter mouse. The expression pattern of the transgene of our line was restricted to cartilage from embryonic to adult stages. β-galactosidase staining was observed in growth plate, articular cartilage, as well as fibrocartilage of meniscus, trachea, and intervertebral discs. Similar staining was observed in a previously described Agc1 (tm(IRES-creERT2)) murine line. The presence of luciferase in our transgene allows the visualization of the transgene expression in live animals. Weekly measurements from 2 to 8 weeks of age showed a reduction in luminescence in knee joints between 2 and 4 weeks of age, but stabilization thereafter. Following the
Lebanon, NH. Dartmouth researchers identify antiviral, survival job descriptions for T cells. Utilizing a novel transgenic mouse model, Edward Usherwood, PhD of Dartmouths Norris Cotton Cancer Center and collaborators found that CD4 T cells divide into two different populations that each has a different job. One type performs antiviral functions, and the other survives life in the host. The study, Functional Heterogeneity in the CD4+ T Cell Response to Murine Y-Herpesvirus 68, was published in the Journal of Immunology.. The human immune response to viruses and cancer is a complex and multi-pronged effort, but in studies like this one, we are making real progress in understanding how to optimize responses against viruses, explained Usherwood.. Gammaherpesviruses such as the Epstein-Barr virus and the Kaposis Sarcoma-associated herpesvirus can cause cancer, mostly in immune-suppressed populations such as patients with AIDS. While immune control of these viruses is believed to rely on ...
Generation of tTA-TRAF2 transgenic mice. To develop an experimental model to study reverse LV remodeling, we generated a conditional transgenic mouse model (tTA-TRAF2) that overexpresses TRAF2 in the heart, using a cardiac-specific and tetracycline-transactivating factor-regulated promoter (23). The rationale for conditionally overexpressing TRAF2 in the heart was that we have shown previously that sustained expression of TRAF2, a scaffolding protein that coordinates signaling through the type 1 and type 2 TNF receptors, resulted in LV remodeling and LV dysfunction that phenocopies the LV remodeling and LV dysfunction observed with the development of parainflammation in mice with cardiac restricted overexpression of TNF (4). Further, elevated levels of myocardial TRAF2 have been reported to be elevated in HF patients (24). In this conditional tet-off system, the stable tetracycline analog dox (Sigma-Aldrich) inhibits tTA transactivation of the Traf2 transgene in the heart.. Briefly, a murine ...
Background: Protons regulate cellular function by modulating the charge and structure of macromolecules, and proton-extruding and importing transport proteins underlie pH homeostasis. The molecular rotary motor F1Fo complex, ATP synthase, was disclosed at the plasma membrane, but its ligands and functions have not been fully understood. We recently identified a circulating peptide coupling factor 6 (CF6), an endogenous prostacyclin inhibitor, as a novel ligand for F1Fo complex: After binding to protrusive F1, CF6 forces the backward rotation of Fo, resulting in proton import. We investigated the role of interaction between CF6 and ecto-F1Fo complex in the genesis of hypertension and diabetes due to tissue acidosis.. Methods and Results: We generated CF6-overexpressing transgenic mouse (TG) in which CF6 was overexpressed by two times compared with wild type mice (WT). In TG, intracellular pH measured by 31P-magnetic resonance spectroscopy was decreased by 0.1 to 0.15 pH unit in the skeletal ...
Double transgenic mice bearing fusion genes consisting of mouse albumin enhancer/promoter-mouse c-myc complementary DNA and mouse metallothionein 1 promoter-human transforming growth factor α complementary DNA were generated to investigate the interaction of these genes in hepatic oncogenesis and to provide a general paradigm for characterizing the interaction of nuclear oncogenes and growth factors in tumorigenesis. Coexpression of c-myc and transforming growth factor α as transgenes in the mouse liver resulted in a tremendous acceleration of neoplastic development in this organ as compared to expression of either of these transgenes alone. The two distinct cellular reactions that occurred in the liver of the double transgenic mice prior to the appearance of liver tumors were dysplastic and apoptotic changes in the existing hepatocytes followed by emergence of multiple focal lesions composed of both hyperplastic and dysplastic cell populations. These observations suggest that the interaction ...