Mouse actin messenger RNAs. Construction and characterization of a recombinant plasmid molecule containing a complementary DNA transcript of mouse alpha-actin mRNA.

Minty, Adrian; Caravatti, M; Robert, Benoît; Cohen, Arlette; Daubas, Philippe; Weydert, A.; Gros, François; Buckingham, M

doi:10.1016/s0021-9258(19)70080-5

Cited by 420 publications

(18 citation statements)

References 48 publications

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“…Total RNA was prepared by acid guanidinium thiocyanatephenol-chloroform extraction (13), and 5-20 ugAample were electrophoresed through formaldehyde/agarose gels . Northern blot analysis was performed as described ( 14) using the IL5 cDNA probe described above and a riboprobe prepared from actin cDNA (15) . Restriction endonucleases and other enzymes were obtained from Boehringer Mannheim (Lewes, UK) or Northumbria Biologicals Ltd. (Cramlington, UK).…”

Section: Methodsmentioning

confidence: 99%

Eosinophilia in transgenic mice expressing interleukin 5.

Dent¹,

Strath²,

Mellor³

et al. 1990

The Journal of Experimental Medicine

543

326

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Experiments in vitro suggest that although interleukin 5 (IL-5) stimulates the late stages of eosinophil differentiation, other cytokines are required for the generation of eosinophil progenitor cells. In this study transgenic mice constitutively expressing the IL-5 gene were established using a genomic fragment of the IL-5 gene coupled to the dominant control region from the gene encoding human CD2. Four independent eosinophilic transgenic lines have thus far been established, two of which with 8 and 49 transgene copies, are described in detail. These mice appeared macroscopically normal apart from splenomegaly. Eosinophils were at least 65- and 265-fold higher in blood from transgenics, relative to normal littermates, and approximately two- or sevenfold more numerous relative to blood from mice infected with the helminth Mesocestoides corti. Much more modest increases in blood neutrophil, lymphocyte, and monocyte numbers were noted in transgenics, relative to normal littermates (less than threefold). Thus IL-5 in vivo is relatively specific for the eosinophil lineage. Large numbers of eosinophils were present in spleen, bone marrow, and peritoneal exudate, and were highest in the line with the greatest transgene copy number. Eosinophilia was also noted in histological sections of transgenic lungs, Peyer's patches, mesenteric lymph nodes, and gut lamina propria but not in other tissues examined. IL-5 was detected in the sera of transgenics at levels comparable to those seen in sera from parasite-infected animals. IL-3 and granulocyte/macrophage colony-stimulating factor (GM-CSF) were not found. IL-5 mRNA was detected in transgenic thymus, Peyer's patches, and superficial lymph nodes, but not in heart, liver, brain, or skeletal muscle or in any tissues from nontransgenics. Bone marrow from transgenic mice was rich in IL-5-dependent eosinophil precursors. These data indicate that induction of the IL-5 gene is sufficient for production of eosinophilia, and that IL-5 can induce the full pathway of eosinophil differentiation. IL-5 may therefore not be restricted in action to the later stages of eosinophil differentiation, as suggested by earlier in vitro studies.

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Section: Methodsmentioning

confidence: 99%

Eosinophilia in transgenic mice expressing interleukin 5.

Dent¹,

Strath²,

Mellor³

et al. 1990

The Journal of Experimental Medicine

543

326

View full text Add to dashboard Cite

show abstract

“…Twenty µg of total RNA was denatured in formaldehyde/formamide, applied to a nylon membrane (Hybond-N + , Amersham) with mild vacuum and hybridized with the W7D probe as for Southern hybridisations (described above). To assess RNA loading, the blot was stripped and reprobed with mouse β-actin cDNA probe (27).…”

Section: Fluorescent In Situ Hybridizationmentioning

confidence: 99%

Xist expression from an Xist YAC transgene carried on the mouse Y chromosome

Matsuura

Episkopou

Hamvas

et al. 1996

Human Molecular Genetics

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We have constructed mouse transgenic lines carrying a YAC clone encompassing the Xist gene in order to investigate the factors influencing Xist expression and the initiation of X-inactivation. Two transgenic lines were derived, one carrying four copies integrated at an autosomal site and a second line carrying four copies integrated at a single site on the Y chromosome. Xist expression was not observed in mice carrying the autosomal insertion. However, Xist expression from the Y-inserted transgenes was observed and at levels commensurate with that found in normal female mice. Methylation sites in the autosomal transgene both 5' and 3' of the Xist gene are hypermethylated and appear to reflect methylation patterns observed on the active X chromosome. For the Y-linked transgene, methylation sites 5' and 3' of the Xist gene are hypomethylated reflecting patterns found on the inactive X chromosome. However, the 5' and 3' methylation levels have been decoupled at the active transgenic locus. The data suggest that sequences in the vicinity of Xist can initiate some of the features that are associated with the initiation process of X-inactivation.

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“…Plasmids were lineariced by restriction enzyme digestion and gel purified to remove uncut templates before transcription, which was done according to Promega recommendations with the following modifications: 12 #m-rUTP, 0.085 #m [32p]rUTP, and 250 #m digoxigenin-11-rUTP (Boehringer Mannheim, GmBH, Mannheim, Germany). The following plasmids were used to make DNA probes for Northern blot analyses: pRK1, containing full-length H-2K d cDNA (see above); pGEM3fl2(A), containing/32-microglobulin (B2m) cDNA, derived from pBReB4 (21) and pAL41 (22), containing murine fl actin cDNA. Inserts were purified by gel electrophoresis and labeled with 32p to a specific activity of 1-2 x 109 dpm/#g, using a random primed DNA labeling kit (Boehringer Mannheim).…”

Section: Infection Of Mic~mentioning

confidence: 99%

Upregulation of class I major histocompatibility complex gene expression in primary sensory neurons, satellite cells, and Schwann cells of mice in response to acute but not latent herpes simplex virus infection in vivo.

Pereira

Tscharke

Simmons

1994

The Journal of Experimental Medicine

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Major histocompatibility complex (MHC) deficiency is typical of almost all resident cells in normal neural tissue. However, CD8+ T cells, which recognize antigenic peptides in the context of class I MHC molecules, are known to mediate clearance of herpes simplex virus (HSV) from spinal ganglia of experimentally infected mice, leading to the hypothesis that class I expression in the peripheral nervous system must be upregulated in response to HSV infection. In addressing this hypothesis it is shown, in BALB/c (H-2d) mice, that normally deficient class I transcripts transiently accumulate in peripheral nerve Schwann cells, ganglionic satellite cells, and primary sensory neurons, indicating that in each of these cell types class I expression is regulated at the transcriptional level in vivo. Furthermore, for 3-4 wk after infection, H-2Kd/Dd antigens are expressed by satellite and Schwann cells but not neurons, suggesting additional posttranscriptional regulation of class I synthesis in neurons. Alternatively, the class I RNAs induced in neurons may not be derived from classical class I genes. Factors regulating H-2 class I expression emanate from within infected ganglia, probably from infected neurons themselves. However, induction of class I molecules was not maintained during latency, when viral gene expression in neurons is restricted to a single region within the virus repeats. These data have implications for the long-term survival of cells in HSV-infected neural tissue.

show abstract

Mouse actin messenger RNAs. Construction and characterization of a recombinant plasmid molecule containing a complementary DNA transcript of mouse alpha-actin mRNA.

Cited by 420 publications

References 48 publications

Eosinophilia in transgenic mice expressing interleukin 5.

Eosinophilia in transgenic mice expressing interleukin 5.

Xist expression from an Xist YAC transgene carried on the mouse Y chromosome

Upregulation of class I major histocompatibility complex gene expression in primary sensory neurons, satellite cells, and Schwann cells of mice in response to acute but not latent herpes simplex virus infection in vivo.

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