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PubMed2006

Comparative Mouse Genomics Centers Consortium: the Mouse Genotype Database.

The Comparative Mouse Genomics Centers Consortium (CMGCC) is a branch of the Environmental Genome Project sponsored by the National Institute of Environmental Health Sciences (NIEHS) focusing upon the identification of human single nucleotide polymorphisms (SNPs) that may confer disease susceptibility within the human population. The goal of the CMGCC (http://www.niehs.nih.gov/cmgcc/) is to make genetic mouse models for human SNPs within cell cycle control, DNA replication and DNA repair genes that may be associated with human pathologies. In order to facilitate information sharing and analysis within the consortium a set of informatics resources have been generated to support the mouse model development efforts. The primary entry point for information about the mouse models developed by the consortium is through the CMGCC Genotype Database (http://mrages.niehs.nih.gov/genotype/), which maintains both a consortium specific and public access display of the available and developing mouse models.

Animals↗
PubMed2006

Immunization with recombinant bovine but not mouse prion protein delays the onset of disease in mice inoculated with a mouse-adapted prion.

Host tolerance to endogenous prion protein (PrP) has hampered the development of prion vaccines as PrP is a major component of prions. Indeed, we show that immunization of mice with mouse recombinant PrP elicited no prophylactic effect against a mouse-adapted prion. However, interestingly, mice immunized with recombinant bovine PrP developed the disease significantly later than non-immunized mice after inoculation of a mouse prion. Sheep recombinant PrP exhibited variable prophylactic effects. Mouse recombinant PrP stimulated only very weak antibody responses. In contrast, bovine recombinant PrP was higher immunogenic and produced variable amounts of anti-mouse PrP autoantibodies. Sheep recombinant PrP was also immunogenic but produced more variable amounts of anti-PrP autoantibodies. These results might open a new way for development of prion vaccines.

Animals↗
PubMed2004

Identification of the mouse H-ficolin gene as a pseudogene and orthology between mouse ficolins A/B and human L-/M-ficolins.

Ficolin is a collagenous lectin which plays a crucial role in innate immunity. Three and two ficolins have been identified in human and mice, respectively. To identify the mouse homologue of human H-ficolin and to elucidate the orthology between mouse ficolins A/B and human L-/M-ficolins, the gene structures were explored. The mouse homologue of the H-ficolin gene was identified as a pseudogene on chromosome 4. The mouse ficolin A gene was located far from the ficolin B gene on chromosome 2, whereas the human L-ficolin and M-ficolin genes were close in the region homologous to the ficolin B locus. Together with the exon-intron structures and the phylogenetic tree, these results suggest that ficolin B is the mouse orthologue of M-ficolin and that the genes encoding serum-type ficolins, ficolin A and L-ficolin, were generated independently from the ficolin B/M-ficolin lineage each in mice and primates.

Amino Acid Sequence↗
PubMed1999

In vitro agonist effects of nociceptin and [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin(1-13)NH(2) in the mouse and rat colon and the mouse vas deferens.

Nociceptin is an endogenous ligand of the opioid receptor-like (ORL1) receptor, a G-protein coupled receptor with sequence similarities to the opioid receptors. ORL1 receptors are present at both central and peripheral sites in several mammalian species but their functions are as yet poorly understood. The main aim of this investigation was to study the effects of nociceptin and the putative ORL1 receptor antagonist [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin(1-13)NH(2) in two peripheral tissues, the isolated proximal colon of the mouse and the distal colon of the rat. Nociceptin, [D-Ala(2), MePhe(4), Gly-ol(5)]enkephalin (DAMGO; mu-opioid receptor selective) and [D-Pen(2), D-Pen(5)]enkephalin (DPDPE; delta-opioid receptor selective) caused concentration-dependent contractions of mouse and rat isolated colon preparations (nociceptin EC(50)=1.20 and 0.28 nM in the mouse and rat, respectively). Des[Phe(1)]nociceptin (250 nM) had no contractile effect. Naloxone (300 nM) antagonised the effects of DAMGO and DPDPE but had no effect in either preparation on contractions seen in response to nociceptin. [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin(1-13)NH(2) also caused contractions in the colonic preparations (EC(50)=6.0 and 3.1 nM in the mouse and rat, respectively); there was no evidence of any antagonist activity. Tetrodotoxin (1 microM) abolished the contractile effects of nociceptin in the mouse colon but had no effect in the rat. In the vas deferens preparation isolated from DBA/2 mice, nociceptin caused concentration-dependent inhibitions of electrically-evoked contractions which were antagonised by [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin(1-13)NH(2) (apparent pK(B)=6. 31). However, [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin(1-13)NH(2) (0.3-10 microM) also possessed agonist activity in this preparation, as it inhibited the electrically-evoked contractions in a concentration-dependent manner. These observations do not support the proposal that [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin(1-13)NH(2) has agonist activity at central ORL1 receptors but is an antagonist in the periphery and that these differences in efficacy point to differences in the receptors. Rather, these data along with those of others suggest that [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin(1-13)NH(2) is a partial agonist and that differences in receptor reserve can account for the varied pharmacological actions of this pseudopeptide at central and peripheral sites.

Animals↗
PubMed2000

Kinetic and genetic bases for the heteroclitic recognition of mouse cytochrome c by mouse anti-pigeon cytochrome c monoclonal antibodies.

The B lymphocyte response to pigeon cytochrome c (CYT) in BALB/c mice was previously shown to initiate as a heteroclitic response specific for the self antigen mouse CYT. As the immune response progressed, the mAb that were produced became less heteroclitic and often bound pigeon CYT with higher affinity than they bound mouse CYT [Minnerath, J. et al., 1995. Proc. Natl. Acad. Sci. USA 92, 12379-12383]. To study the basis for heterocliticity and its loss in this system, the H and L chain amino acid sequences of anti-pigeon CYT mAb obtained from the primary and secondary Ab responses were first compared. The most frequent somatic mutations and Ig gene joints were then introduced into an engineered single-chain Fv (scFv) that expressed the germline-encoded V(H) and V(L) amino acid sequences. The effects of those changes on the on- rate, off-rate, and affinity constants in binding both mouse and pigeon CYT were determined by surface plasmon resonance. In this system, the heterocliticity of primary mAb was largely due to a decreased on-rate constant for binding pigeon CYT relative to mouse CYT. Various combinations of the three frequently occurring H chain somatic mutations (H31, H56, and H58) increased the on-rate constant to maximal levels. Only one of the mutations (H58) decreased the off-rate constant when in combination with the other mutations and the effect of H58 was greater for scFv binding mouse CYT than pigeon CYT. Consequently, the mutated scFv and many secondary mAb remained heteroclitic, although their affinities for pigeon CYT increased. Secondary mAb that were no longer heteroclitic expressed non-canonical amino acid sequences in the V(H)-D-J(H) joint in the context of the canonical V genes or expressed different V genes. In addition to providing insight into the molecular basis for heterocliticity, our findings confirm that both faster on-rate and slower off-rate constants are favored during affinity maturation of the Ab response.

Amino Acid Sequence↗
PubMed1992

Rat thymic epithelium positively selects mouse T cells with specificity for rat MHC class II antigens but fails to induce detectable tolerance in the mouse T cells to the rat MHC antigens.

BALB/c (H-2d) nude mice were grafted with allogeneic AKR/J (H-2k) or xenogeneic (ACI-N rat, RT1av1) fetal thymuses which were depleted of hemopoietic cells by incubating with 2'-deoxyguanosine (2'dGuo) in vitro prior to grafting. The nylon-wool-passed LN T cells from nude mice grafted with 2'dGuo-treated AKR/J thymus showed a poor proliferative response to B10BR (H-2k) stimulator cells, confirming that mouse thymic epithelium has the capacity to induce tolerance against the mouse MHC antigens on the thymic epithelium. On the other hand, the nylon-wool-passed LN T cells from nude mice grafted with untreated or 2'dGuo-treated ACI/N rat thymus showed significant proliferative responses to ACI/N, which can be blocked by anti-rat MHC class II mAb, whereas the nylon-wool-passed LN T cells from nude mice grafted with syngeneic thymus hardly responded to the xenogeneic stimulator cells. These results suggest that rat thymic stromal cells including thymic epithelium can not induce detectable tolerance in mouse T cells to rat MHC antigens; but rat thymic epithelium may positively select mouse T cells with specificity for rat MHC class II antigens, resulting in a mouse T cell repertoire with strong xeno-reactivity.

Animals↗
PubMed1998

Opioid peptide receptor studies. 8. One of the mouse brain deltaNCX binding sites is similar to the cloned mouse opioid delta receptor: further evidence for heterogeneity of delta opioid receptors.

Quantitative ligand binding studies resolved two subtypes of the delta opioid receptor, termed delta(ncx1) and delta(ncx2), in mouse brain membranes depleted of mu receptors by pretreatment with the irreversible ligand, BIT. The purpose of the present study was to compare the binding parameters, ligand-selectivity profile and pharmacological properties of the cloned mouse delta receptor (MDOR) stably expressed in a cell line to the delta(ncx) binding sites of mouse brain. [3H][D-Ala2,D-Leu5]enkephalin labeled a single binding site in membranes prepared from MDOR cells under several different assay conditions including BIT-pretreatment. The MDOR had high affinity for delta agonists and antagonists. [3H][D-Ala2,D-Leu5]enkephalin labeled two binding sites in mouse brain membranes depleted of mu receptors by pretreatment with BIT: the delta(ncx1) site (high affinity for DPDPE and deltorphin) and the delta(ncx2) site (low affinity for DPDPE and deltorphin). Some agents were moderately selective for the delta(ncx2) site: [pCl]DPDPE (10.9-fold), JP41 (5.9-fold) and JP45 (3.8-fold). The Ki values of 12 opioids at the mouse MDOR were determined. These values were highly correlated with their values at the delta(ncx1) site but not the delta(ncx2) site. These data suggest that the delta(ncx2) site may be distinct from the cloned delta opioid receptor.

Animals↗
PubMed2000

Characterization of the mouse Smad1 gene and its expression pattern in adult mouse tissues.

Smad1 belongs to a family of receptor-activated proteins which mediate signals from TGF-beta superfamily ligands, including TGF-beta and BMPs. Although much is known about the biochemistry of Smad1 signal transduction, the role of Smad1 in vivo is still unclear. Here we present the first description of the genomic structure of the mouse Smad1 gene and the characterization of its expression pattern in adult mouse tissues by immunohistochemistry. The Smad1 gene contains 7 exons and spans >42 kb of genomic DNA. Its coding region is contained within 6 exons and all introns, except intron 1, follow the GT/AG rule. Immunohistochemical analysis shows that Smad1 is widely expressed in adult mouse tissues, with a varying degree of nuclear localization in different cell types, suggesting a regulated function for this protein. This study assigns all of the exon-intron boundaries of the mouse Smad1 gene and provides the basis for assessing the functional significance of this gene using targeted gene manipulation in the mouse.

5' Untranslated Regions↗
PubMed2001

Isolation and characterization of mouse MUC18 cDNA gene, and correlation of MUC18 expression in mouse melanoma cell lines with metastatic ability.

The cell surface adhesion molecule human MUC18 (huMUC18 or Mel-CAM) has been postulated to play a key pathogenic role in metastatic melanoma progression. To establish an immunocompetent syngeneic mouse model that would greatly facilitate our understanding of the role of MUC18 in the metastatic behavior of melanoma, we cloned and characterized the mouse MUC18 (muMUC18) cDNA gene. The gene was amplified by RT-PCR and RACE of the poly(A)+RNA isolated from the mouse melanoma cell line B16F10/Queens. The cloned muMUC18 cDNA gene contained 28 nucleotides of 5'-UTR, 908 nucleotides of 3'-UTR, and an open reading frame (ORF) of 1947 nucleotides encoding a protein of 648 amino acids, which is two amino acids longer than huMUC18. The size of the muMUC18 mRNA is about 3 kb with a shorter 3'-UTR than the huMUC18 mRNA (about 3.3 kb). Besides, the sequence in the 3' UTR of the two mRNAs is diverse with only 31% identity. The 5'-UTR and coding sequences of the muMUC18 cDNA are 72.4 and 80.6% identical to those of huMUC18, respectively. The deduced amino acid sequence of the muMUC18 cDNA is 76.2% identical to that of huMUC18. The amino acid sequences deduced from MUC18 cDNA sequences from six other mouse melanoma cell lines are identical except one to three residues, suggesting that the muMUC18 cDNA sequence determined in this report is correct. The muMUC18 protein is predicted to be slightly more acidic than the human protein. The levels of muMUC18 mRNA and protein in nine mouse melanoma cell lines were directly proportional to their ability to establish metastatic colonies in lungs of syngeneic mice. Most biological functions of the muMUC18 may be similar to the huMUC18.

Amino Acid Sequence↗
PubMed1997

The mouse uracil-DNA glycosylase gene: isolation of cDNA and genomic clones and mapping ung to mouse chromosome 5.

Uracil-DNA glycosylase (UDG) is the enzyme responsible for the first step in the base-excision repair pathway that specifically removes uracil from DNA. Here we report the isolation of the cDNA and genomic clones for the mouse uracil-DNA glycosylase gene (ung) homologous to the major placental uracil-DNA glycosylase gene (UNG) of humans. The complete characterization of the genomic organization of the mouse uracil-DNA glycosylase gene shows that the entire mRNA coding region for the 1.83-kb cDNA of the mouse ung gene is contained in an 8.2-kb SstI genomic fragment which includes six exons and five introns. The cDNA encodes a predicted uracil-DNA glycosylase (UDG) protein of 295 amino acids (33 kDa) that is highly similar to a group of UDGs that have been isolated from a wide variety of organisms. The mouse ung gene has been mapped to mouse chromosome 5 using fluorescence in situ hybridization (FISH).

Amino Acid Sequence↗
PubMed2003

Molecular analysis of human Siglec-8 orthologs relevant to mouse eosinophils: identification of mouse orthologs of Siglec-5 (mSiglec-F) and Siglec-10 (mSiglec-G).

We recently identified a novel human sialic acid binding immunoglobulin-like lectin, Siglec-8, using mRNA from human eosinophils. To search for a mouse Siglec (mSiglec) ortholog of Siglec-8 and other mouse Siglec paralogs, we conducted public database searches with cDNA sequences of human Siglec-5 to -10 and identified two novel mSiglecs. One has significant sequence identity to human Siglec-5 and is a splice variant of mSiglec-F. The other has greatest sequence identity to human Siglec-10 (mSiglec-G). Both mSiglecs have extracellular Ig-like domains and intracellular tyrosine-based motifs. To determine whether these mSiglecs were relevant to mouse eosinophils, RT-PCR and Northern blot analysis were performed. We detected expression of mSiglec-5 (or -F), -10, and -E mRNA in purified mouse eosinophils, but Northern blot data comparing expression in tissues from normal, IL-5 transgenic, and allergen-sensitized and -challenged mice suggest that mSiglec-10 is probably most relevant to mouse eosinophils.

Amino Acid Sequence↗
PubMed2003

Characterization of HLA DR2 and DQ8 transgenic mouse with a new engineered mouse class II deletion, which lacks all endogenous class II genes.

Human autoimmune diseases are a class of complex immune system disorders characterized by loss of tolerance to self-antigens. HLA class II molecules play a central role in the initiation, propagation and prolongation of the disease process. HLA class II transgenic mice with mouse endogenous class II gene Ab knockout were used successfully in several mouse models for human autoimmune diseases, such as IDDM, SLE and EAE in our Lab. However, these mice carry the functional mouse Eb gene from the Abeta(0/0) construct and could express Ebeta/DRalpha(Ealpha) molecules and shape the T cell repertoire in these mice. Recently, we have obtained the new MHCII(Delta/Delta) mice that are devoid of all endogenous conventional mouse MHC class II genes. When these mice are mated with our HLA class II transgenic mice, only human class II genes are expressed. The DR and DQ molecules expressed in these mice shape the T cell repertoire and regulate the immune response. Therefore, this new class of HLA transgenic mice is the first to be completely "humanized" in their MHC class II genes and will be an invaluable mouse model for human MHC class II associated autoimmune diseases.

Animals↗
PubMed2003

Identification of chromosome rearrangements between the laboratory mouse (Mus musculus) and the Indian spiny mouse (Mus platythrix) by comparative FISH analysis.

Comparative chromosome painting was applied to the Indian spiny mouse (Mus platythrix) with mouse (M. musculus) chromosome-specific probes for understanding the process of chromosome rearrangements between the two species. The chromosome locations of the 5S and 18S-28S ribosomal RNA genes and the order of the 119 and Tcp-1 genes in the In(17)2 region of the t-complex were also compared. All the painting probes were successfully hybridized to the Indian spiny mouse chromosomes, and a total of 27 segments homologous to mouse chromosomes were identified. The comparative FISH analysis revealed that tandem fusions were major events in the chromosome evolution of the Indian spiny mouse. In addition, other types of chromosome rearrangements, i.e. reciprocal translocations and insertions, were also included.

Animals↗
PubMed2000

A three-dimensional skin culture model for mouse keratinocytes: application to transgenic mouse keratinocytes.

The study of mouse epidermal biology has been hampered by the lack of a good in vitro model for the culture of mouse keratinocytes which allowed the reconstruction of a fully differentiated epidermis. We adapted the Pruniéras' model, also called the Dead de-Epidermized Dermis model (DED), to mouse keratinocytes and showed that a neo-epidermis can be reconstructed exhibiting a complete differentiation program. We also used this model to culture transgenic mouse keratinocytes. We observed that transgene expression occurred in the correct location and that the neo-epidermis mimed previous in vivo observations obtained with integrin skin-targeted transgenic mice. Therefore, this model will be a powerful tool to further investigate normal mouse and transgenic keratinocyte biology.

Animals↗
PubMed1993

Syrian hamster embryo cell lines useful for detecting transforming genes in mouse tumours: detection of transforming genes in X-ray-related mouse tumours.

The Syrian hamster embryo cell lines, SHOK and MC-1, were used as recipient cells for DNA transfection assay to detect transforming genes in experimental mouse tumours. A mouse repeat sequence was utilised to check whether each transformed focus included mouse genomic DNA in the Hamster background. We investigated five mouse tumours that are related to X-ray radiation, and detected activated c-K-ras, c-mos, and c-cot oncogenes which induced foci of hamster cells. These results show that SHOK and MC-1 cells have unique properties for detecting transforming genes in experimental mouse tumours.

3T3 Cells↗
PubMed1974

Differences in the products of mitochondrial protein synthesis in vivo in Human and mouse cells and their potential use as markers for the mitochondrial genome in human--mouse somatic cell hybrids.

The proteins synthesized in the mitochondria of mouse and human cells grown in tissue culture were examined by electrophoresis in polyacrylamide gels. The proteins were labelled by incubating the cells in the presence of [(35)S]methionine and an inhibitor of cytoplasmic protein synthesis (emetine or cycloheximide). A detailed comparison between the labelled products of mouse and human mitochondrial protein synthesis was made possible by developing radioautograms after exposure to slab-electrophoresis gels. Patterns obtained for different cell types of the same species were extremely similar, whereas reproducible differences were observed on comparison of the profiles obtained for mouse and human cells. Four human-mouse somatic cell hybrids were examined, and in each one only components corresponding to mouse mitochondrially synthesized proteins were detected.

Animals↗
PubMed1984

Effect of human seminal plasma and mouse accessory gland extracts on mouse fertilization in vitro.

The mouse in vitro fertilization system was used to investigate the effect of human seminal plasma (HSP) on the fertilizing ability of mouse spermatozoa. The addition of HSP to freshly collected mouse epididymal spermatozoa decreased their fertilizing ability to 30%, compared with 85-90% for control spermatozoa not exposed to HSP or for spermatozoa that had been exposed to the capacitating system for 55-145 min (which allowed capacitation to occur before the addition of HSP). Human seminal plasma from a vasectomized donor was more effective in retarding the acquisition of fertilizing ability in mouse spermatozoa than was seminal plasma from non-vasectomized donors. Fluid from the prostate glands and seminal vesicles of mice also reduced the fertilizing ability of freshly collected mouse spermatozoa to 30%.

Animals↗
PubMed1972

Expression of differentiated functions in hepatoma cell hybrids: induction of mouse albumin production in rat hepatoma-mouse fibroblast hybrids.

The synthesis of serum albumin has been studied in hybrids between well-differentiated rat hepatoma cells, which synthesize serum albumin, and mouse fibroblasts (3T3) that do not synthesize albumin. By immunodiffusion techniques with noncrossreacting antisera, the production of both rat and mouse albumin by the hybrids has been examined. Karyologically identified hybrids were produced between 3T3 cells and cells of a 1s hepatoma (Fu5) clone, and of a 2s hepatoma (2s Fu5-5cl.lE) clone. Each of the 3T3 x Fu5 hybrids produces only rat albumin. Among five 3T3 x 2s Fu5-5cl.lE hybrid clones isolated, one produces both rat and mouse albumin, two produce only mouse albumin, and two do not produce rat or mouse albumin.

Animals↗