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At least 397 records · Page 22Linked to original sources
PubMed1994

Long-term impaired neutrophil migration in mice overexpressing human interleukin-8.

The proinflammatory chemokine interleukin-8 (IL-8/NAP-1) has been implicated in recruiting neutrophils to sites of acute and chronic tissue inflammation. In transgenic mice, elevated serum IL-8 levels ranging from 1 to 118 ng/ml were correlated with proportional increases in circulating neutrophils and proportional decreases in L-selectin expression on the surface of blood neutrophils. No change in the expression of the beta 2-integrins Mac-1 and LFA-1 was apparent on peripheral blood neutrophils of the IL-8 transgenic mice. Additionally, L-selectin expression on bone marrow neutrophils and neutrophil precursors was normal in all transgenic lines. IL-8 transgenic mice demonstrated an accumulation of neutrophils in the microcirculation of the lung, liver and spleen. Moreover, there was no evidence of neutrophil extravasation, plasma exudation or tissue damage in any IL-8 transgenic mice. Neutrophil migration into the inflamed peritoneal cavity was severely inhibited in IL-8 transgenic mice, but not in nontransgenic littermates. The IL-8 transgenic mice should serve as useful models for studying the putative role of neutrophils in mediating tissue damage in models of inflammation, such as hepatic ischemia and reperfusion injury, cecal puncture and ligation, and glomerulonephritis.

Animals↗
PubMed1999

OCD-Like behaviors caused by a neuropotentiating transgene targeted to cortical and limbic D1+ neurons.

To study the behavioral role of neurons containing the D1 dopamine receptor (D1+), we have used a genetic neurostimulatory approach. We generated transgenic mice that express an intracellular form of cholera toxin (CT), a neuropotentiating enzyme that chronically activates stimulatory G-protein (Gs) signal transduction and cAMP synthesis, under the control of the D1 promoter. Because the D1 promoter, like other CNS-expressed promoters, confers transgene expression that is regionally restricted to different D1+ CNS subsets in different transgenic lines, we observed distinct but related psychomotor disorders in different D1CT-expressing founders. In a D1CT line in which transgene expression was restricted to the following D1+ CNS regions-the piriform cortex layer II, layers II-III of somatosensory cortical areas, and the intercalated nucleus of the amygdala-D1CT mice showed normal CNS and D1+ neural architecture but increased cAMP content in whole extracts of the piriform and somatosensory cortex. These mice also exhibited a constellation of compulsive behavioral abnormalities that strongly resembled human cortical-limbic-induced compulsive disorders such as obsessive-compulsive disorder (OCD). These compulsive behaviors included episodes of perseverance or repetition of any and all normal behaviors, repetitive nonaggressive biting of siblings during grooming, and repetitive leaping. These results suggest that chronic potentiation of cortical and limbic D1+ neurons thought to induce glutamatergic output to the striatum causes behaviors reminiscent of those in human cortical-limbic-induced compulsive disorders.

Aggression↗
PubMed1996

Decreased Fas antigen receptor expression in testicular tumor cell lines derived from polyomavirus large T-antigen transgenic mice.

MT-PVLT-10 transgenic mice express large T-antigen of polyomavirus under the control of the mouse metallothionein-1 promoter and mates of this transgenic line develop testicular tumors at advanced ages. The differential display technique was employed to compare mRNA expression from immortalized cell lines derived from normal or adenomatous testis from MT-PVLT-10 transgenic males. Using this technique, a complementary DNA fragment corresponding to the mouse Fas antigen receptor was recovered from normal testicular cells but not from tumor cells. RNAse protection assays with the Fas antigen specific fragment confirmed its differential expression. Normal testicular cells from the transgenic animals responded to treatment of interferon-gamma by increasing the expression of Fas antigen specific mRNA and were sensitive to the proliferative inhibitory effect of anti-Fas antibody in vitro. This proliferative inhibition was characterized by an accumulation of cells in S phase of the cell cycle. In contrast, the testicular tumor cells did not respond to either interferon-gamma or to anti-Fas antibody in vitro. These results suggest that the toss of proliferative inhibitory effect mediated by the Fas antigen pathway in tumor cells may be an important step in testicular tumor progression in the MT-PVLT-10 transgenic mice.

Animals↗
PubMed1998

Transgenic mice for MTCP1 develop T-cell prolymphocytic leukemia.

T-cell prolymphocytic leukemia (T-PLL) is a rare form of mature T-cell leukemia associated with chromosomal rearrangements implicating MTCP1 or TCL1 genes. These genes encode two homologous proteins, p13(MTCP1) and p14(TCL1), which share no similarity with other known protein. To determine the oncogenic role of MTCP1, mice transgenic for MTCP1 under the control of CD2 regulatory regions (CD2-p13 mice) were generated. No abnormality was detected during the first year after birth. A late effect of the transgene was searched for in a cohort of 48 CD2-p13 mice aged 15 to 20 months, issued from 3 independent founders. Lymphoid hemopathies, occurring in the three transgenic lines, were characterized by lymphoid cells with an irregular nucleus, a unique and prominent nucleolus, condensed chromatin, a basophilic cytoplasm devoid of granules, and an immunophenotype of mature T cells. The molecular characterization of Tcrb rearrangements demonstrated the monoclonal origin of these populations. Histopathological analysis of the cohort demonstrated early splenic and hepatic infiltrations, whereas lymphocytosis and medullar infiltrations were found infrequently. The engraftment of these proliferations in H2-matched animals demonstrated their malignant nature. Cumulative incidence of the disease at 20 months was 100%, 50%, and 21% in F3, F4, and F7 lines, respectively, and null in the control group. The level of expression of the transgene, as estimated by Western blotting in the transgenic lines correlated with the tumoral incidence, with the highest expression of p13(MTCP1) being found in F3 mice. CD2-p13 transgenic mice developed an hemopathy similar to human T-PLL. These data demonstrate that p13(MTCP1) is an oncoprotein and that CD2-p13 transgenic mice represent the first animal model for mature T-PLL.

Animals↗
PubMed1992

Harderian gland hyperplasia in c-mos transgenic mice.

Transgenic mice carrying the mouse mos proto-oncogene linked to a retroviral LTR develop hyperplasia of the Harderian glands. Enlargement of the glands is evident as early as 18 weeks after birth, with glands reaching up to 10 times their normal weight. Approximately 65% of the cases of hyperplasia occur bilaterally, and the majority of mice affected are male (66%). Elevated levels of mos expression are found in all Harderian glands of mice from the affected transgenic line, but not in glands of normal mice or a non-affected transgenic line, indicating that hyperplasia is dependent on mos expression. Histological examination of the tissue reveals a general involvement of the entire gland epithelium in hyperplastic growth, with no evidence of focal or malignant tumours. These observations show that in addition to neu, myc, ras and ret transgenes, mos, a member of the protein-serine/threonine kinase family of oncogenes, can induce Harderian gland hyperplasia, thus revealing an unusual response by this organ to various classes of oncogenes. Analysis of fos, jun, myc and ets oncogene RNA in mos-induced hyperplastic Harderian glands shows that there are no consistent changes in the level of expression of these oncogenes, suggesting that mos acts via a mechanism other than by increasing the expression of these genes.

Animals↗
PubMed2000

Expression of a Petunia inflata pectin methyl esterase in Solanum tuberosum L. enhances stem elongation and modifies cation distribution.

Transgenic potato (Solanum tuberosum L.) plants were constructed with a Petunia inflata-derived cDNA encoding a pectin methyl esterase (PME; EC 3.1.1.11) in sense orientation under the control of the cauliflower mosaic virus 35S promoter. The PME activity was elevated in leaves and tubers of the transgenic lines but slightly reduced in apical segments of stems from mature plants. Stem segments from the base of juvenile PME-overexpressing plants did not differ in PME activity from the control, whereas in apical parts PME was less active than in the wild-type. During the early stages of development stems of these transgenic plants elongated more rapidly than those of the wild-type. Further evidence that overexpression of a plant-derived PME has an impact on plant development is based on modifications of tuber yield, which was reduced in the transgenic lines. Cell walls from transgenic tubers showed significant differences in their cation-binding properties in comparison with the wild-type. In particular, cell walls displayed increased affinity for sodium and calcium, while potassium binding was constant. Furthermore, the total ion content of transgenic potatoes was modified. Indications of PME-mediated differences in the distribution of ions in transgenic plants were also obtained by monitoring relaxations of the membrane potential of roots subsequent to changes in the ionic composition of the bathing solution. However, no effects on the chemical structure of pectin from tuber cell walls could be detected.

Carboxylic Ester Hydrolases↗
PubMed2002

Germ line transformation of the yellow fever mosquito, Aedes aegypti, mediated by transpositional insertion of a piggyBac vector.

Mosquito-vectored diseases such as yellow fever and dengue fever continue to have a substantial impact on human populations world-wide. Novel strategies for control of these mosquito vectored diseases can arise through the development of reliable systems for genetic manipulation of the insect vector. A piggyBac vector marked with the Drosophila melanogaster cinnabar (cn) gene was used to transform the white-eyed khw strain of Aedes aegypti. Microinjection of preblastoderm embryos resulted in four families of cinnabar transformed insects. An overall transformation frequency of 4%, with a range of 0% to as high as 13% for individual experiments, was achieved when using a heat-shock induced transposase providing helper plasmid. Southern hybridizations indicated multiple insertion events in three of four transgenic lines, while the presence of duplicated target TTAA sites at either ends of individual insertions confirmed characteristic piggyBac transposition events in these three transgenic lines. The transgenic phenotype has remained stable for more than twenty generations. The transformations effected using the piggyBac element establish the potential of this element as a germ-line transformation vector for Aedine mosquitoes.

Aedes↗
PubMed2003

[The viral resistance of transgenic tobacco plants containing untranslatable PVYN coat protein gene].

In previous study, we have proved that the resistance of the transgenic tobacco plants containing untranslatable PVYN CP gene was mediated by PVYN CP transgene RNAs, and the resistance mechanism was similar to PTGS. In this paper, T0 progeny transgenic lines with different resistant levels were selected to further study the transgene inheritance and resistant stability of transgenic plants. Results showed that T0 progeny susceptible lines, which contained 1-2 transgene copies, displayed a 3:1 segregation ratio in T1 progeny lines; T0 progeny middle resistant lines, which contained 3-4 transgene copies, revealed a 15:1 segergation ratio in T1 progeny lines; T0 progeny highly resistant lines, which contained 5-7 transgene copies, followed 15:1 or 63:1 segregation pattern in T1 progeny lines. Southern blot analysis revealed the resistance in most T1 and T2 progeny transgenic lines was related to copy numbers of the transgene. Northern blot analysis indicated that PVYN CP transgenes were expressed at transcription level in most T1 and T2 progeny transgenic lines, and the transgene mRNA accumulation in cytoplasm varied among transgenic lines. There was an inverse correlation between transgene transcript accumulation and virus resistance. Resistance of transgenic lines was inheritable over at least two generations, and the resistance of T2 progeny transgenic lines, which containing untranslatable PVYN CP gene, was (1) not overcome by PVYN particle or PVYN RNA; (2) independent of inoculum levels; (3) resistant to either aphid or mechanically transmitted PVYN; (4) not dependent on plant development stages; (5) PVY-specific (i.e., broad-spectrum resistance was not observed).

Capsid Proteins↗
PubMed1993

Establishment of gastric surface mucous cell lines from transgenic mice harboring temperature-sensitive simian virus 40 large T-antigen gene.

We succeeded in establishing two gastric surface mucous cell lines (designated as GSM06 and GSM10), which produce periodic acid-Schiff (PAS)- and concanavalin A (Con A)-positive glycoproteins, from a primary culture of gastric fundic mucosal cells of adult transgenic mice harboring a temperature-sensitive simian virus 40 large T-antigen gene. At the permissive temperature (33 degrees C), GSM06 and GSM10 cells grew until confluent monolayers were formed and have now been in culture for more than 9 months with regular passaging. Con A-horseradish peroxidase- and PAS-positive staining indicated that these cells retain the characteristics of gastric surface mucous cells. GSM06 cells showed temperature-sensitive growth in culture and expressed SV40 large T-antigen at the permissive temperature but not at a nonpermissive temperature (39 degrees C). In contrast, GSM10 cells showed temperature-insensitive growth and expressed T-antigen at both permissive and nonpermissive temperatures. To our knowledge, this is the first report of the establishment of gastric surface mucous cell lines from animals. These immortalized cell lines with normal characteristics may serve as good experimental models for the basic and applied biology of gastric surface mucous cells.

Animals↗
PubMed2005

A microarray-based comparative analysis of gene expression profiles during grain development in transgenic and wild type wheat.

Global, comparative gene expression analysis is potentially a very powerful tool in the safety assessment of transgenic plants since it allows for the detection of differences in gene expression patterns between a transgenic line and the mother variety. In the present study, we compared the gene expression profile in developing seeds of wild type wheat and wheat transformed for endosperm-specific expression of an Aspergillus fumigatus phytase. High-level expression of the phytase gene was ensured by codon modification towards the prevalent codon usage of wheat genes and by using the wheat 1DX5HMW glutenin promoter for driving transgene expression. A 9K wheat unigene cDNA microarray was produced from cDNA libraries prepared mainly from developing wheat seed. The arrays were hybridised to flourescently labelled cDNA prepared from developing seeds of the transgenic wheat line and the mother variety, Bobwhite, at three developmental stages. Comparisons and statistical analyses of the gene expression profiles of the transgenic line vs. that of the mother line revealed only slight differences at the three developmental stages. In the few cases where differential expression was indicated by the statistical analysis it was primarily genes that were strongly expressed over a shorter interval of seed development such as genes encoding storage proteins. Accordingly, we interpret these differences in gene expression levels to result from minor asynchrony in seed development between the transgenic line and the mother line. In support of this, real time PCR validation of results from selected genes at the late developmental stage could not confirm differential expression of these genes. We conclude that the expression of the codon-modified A. fumigatus phytase gene in the wheat seed had no significant effects on the overall gene expression patterns in the developing seed.

6-Phytase↗
PubMed2006

[Raising fat content in transgenic rice by anti-PEP gene transformation].

Gene for phosphoenolpyruvate carboxylase (PEPCase) was incorporated in antisense orientation into genome of japonica rice variety Xiushui 11 through Agrobacterium tumefaciens-mediated transformation of mature seeds-derived embryogenic calli, and a total of 95 transgenic plants were regenerated, which were confirmed by histochemical detection of GUS activity (Fig.2), PCR assay (Fig.3) and Southern hybridization analysis (Fig.4). The transgene was inherited by the T1 progeny and Mendelian 3:1 segregation ratio was observed in most transgenic lines. It was found that the fat content of hulled grains of T2 transgenic lines was 0.37 +/- 0.12 per cent higher than that of the control, most of which were statistically very significant (Table 2).

Agrobacterium tumefaciens↗
PubMed2001

Rescue of a transgenic mouse line by transplantation of a frozen-thawed ovary obtained postmortem.

During the course of breeding valuable mutant or transgenic mice, deaths sometimes occur due to sudden-onset disease or accident. We previously showed that mice can be rescued by transplantation of ovaries taken up to 2 h after death from dead mice remaining at conditions of constant temperature (22 +/- 2 degrees C) and humidity (55% +/- 5%). To extend the flexibility of transplantation, we assessed whether it is possible to cryopreserve ovaries taken from dead mice within 2 h after death. Fertile transgenic mice used as donors were euthanized by cervical dislocation and left for 2 h after death. The cryopreservation was based on Sztein's method with a controlled-rate freezer or on Rall and Fahy's method without a controlled-rate freezer. The recipient mice were nontransgenic littermates of the donor mice, and after transplantation of the frozen-thawed ovaries, they were mated with proven-fertile males. Polymerase chain reaction (PCR) analysis confirmed that the progeny carried the transgene. We show here that by using both of the described methods, it is possible to cryopreserve the ovaries taken from dead mice within 2 h after death and that the mice into which the cryopreserved ovaries are transplanted are fertile.

Animals↗
PubMed2006

HuC:Kaede, a useful tool to label neural morphologies in networks in vivo.

A simple and efficient procedure for labeling neurons is a prerequisite for investigating the development of neural networks in zebrafish. To label neurons we used Kaede, a fluorescent protein with a photoconversion property allowing conversion from green to red fluorescence following irradiation with UV or violet light. We established a zebrafish stable transgenic line, Tg(HuC:Kaede), expressing Kaede in neurons under the control of the HuC promoter. This transgenic line was used to label a small number of neurons in the trigeminal ganglion. Also, using embryos injected with the transgene, we labeled peripheral axon arbors of a Rohon-Beard neuron at 4 days postfertilization and observed the dendrite development of a tectal neuron for 3 days. These data indicate that Kaede is a useful tool to selectively label neural networks in zebrafish.

Animals↗
PubMed2005

Delay in flowering and increase in biomass of transgenic tobacco expressing the Arabidopsis floral repressor gene FLOWERING LOCUS C.

FLOWERING LOCUS C (FLC), a gene from Arabidopsis thaliana (L.) Heynh. that acts as a flowering repressor, was expressed in tobacco (Nicotiana tabacum L. 'Samsun'). Five putative transgenic lines were selected and examined for the presence of FLC. Genomic DNA and total RNA were isolated from the Leaves and used for polymerase chain reaction (PCR) and RNA blot analysis, respectively. Both DNA and RNA tests confirmed the integration and transcription of FLC in all five Lines and their T1 progenies. Transgenic plants in one Line showed an average of 36 d delay in flowering time compared to control plants, and the overall mean for all lines was 14 d. Transgenic plants also displayed increased leaf size and biomass yield and reduced height at flowering time. It is important to note that the delay in flowering might have been caused by a slower rate of leaf initiation (i.e. nodes/day) rather than by a change in the flowering mechanism itself.

Arabidopsis Proteins↗
PubMed2001

Co-expression of multiple transgenes in mouse CNS: a comparison of strategies.

The introduction of two transgenes into one animal is increasingly common as transgenic experiments become more sophisticated. In this study we examine two strategies for creating double transgenic founders from a single microinjection. In the first approach, two constructs, each with its own promoter element, were coinjected into the pronucleus. In the second approach, both transgenes were cloned into one vector, separated by an internal ribosomal entry site (IRES), and placed under control of a single promoter. Both strategies save time and increase the percentage of double transgenic offspring over the standard method of mating single transgenic lines. However, despite high transgene copy numbers, the bicistronic lines did not show robust expression of either protein. Copy number and protein expression correlated much better in the coinjected lines, with expression levels in one line approaching that observed in some of our best single transgenic controls. Thus we recommend coinjection of individual plasmids for the generation of multiply transgenic founders.

Amyloid beta-Protein Precursor↗
PubMed1997

Novel smooth muscle cell lines from transgenic mice harboring temperature-sensitive SV40 large T-antigen gene. Temperature-dependent expression of smooth muscle myosin heavy chain-1 and calponin genes.

We have established novel vascular smooth muscle cell lines (SVS30 and SVS24 cells) which retain the expression of specific markers for smooth muscle cells, such as alpha-actin, smooth muscle myosin heavy chain-1, and calponin, from transgenic mice harboring the temperature-sensitive SV40 large T-antigen gene. SVS cell lines showed temperature-dependent growth and the expression of SV40 large T-antigen. Interestingly, protein and mRNA levels of smooth muscle myosin heavy chain-1 and calponin seen in culture at the non-permissive temperature (39 degrees C) were higher than those at the permissive temperature (33 degrees C). These results suggest that SV40 large T-antigen affects the expression of smooth muscle-specific markers in SVS cell lines, and that some of the characters in SVS cell lines can be controlled by culture temperature. SVS cell lines should be quite valuable tools with which to study the regulation of phenotypic modulation of smooth muscle cells, and to identify smooth muscle specific transcription factors which involve the expression of smooth muscle myosin heavy chain-1 and calponin genes.

Animals↗
PubMed2006

0.2kb promoter sequence of the murine Cyp19 gene target beta-galactosidase expression to specific brain areas of transgenic mice.

Cyp19 encodes the key enzyme of estrogen biosynthesis, aromatase cytochrome P450. In mice it is mainly expressed in the ovary and brain, where transcription is directed by a distal, brain-specific promoter (P(br)). In order to map functional sequence elements of P(br), portions of various length (0.2, 1.0, and 1.7[kb]) were fused to a lacZ reporter gene and analyzed in transgenic mice. Numbers of integrated reporter genes varied from 1 to 23 copies in different transgenic lines. These copy numbers however did not show any correlation to the levels of transgene expression. All of the constructs were found being expressed in the olfactory bulb, limbic cortex, amygdala, and hypothalamus. Additional expression in thalamic nuclei, bed nucleus of stria terminalis, and dorsal mesencephalon was found in transgenic lines with constructs 1.0 and 1.7, and expression in septal and preoptic nuclei was only found with construct 1.7. The data demonstrate that 0.2kb of P(br) target reporter gene expression to specific brain areas. The data also strongly suggest that the sequence between 0.2 and 1.7kb upstream, is necessary for expression in additional areas. However even 1.7kb of P(br) are not sufficient to consistently mimic the accurate expression pattern of Cyp19.

Animals↗