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At least 649 records · Page 36Linked to original sources
PubMed2000

Comparison of oocyte activation and Ca2+ oscillation-inducing abilities of round/elongated spermatids of mouse, hamster, rat, rabbit and human assessed by mouse oocyte activation assay.

Oocyte activation and Ca2+ oscillation-inducing abilities of round spermatid (ROS) and elongated spermatid (ELS) of some rodents and human were assessed by their injection into mouse (B6D2F1) oocytes (mouse test). With mice (B6D2F1, ICR) and rat, ROS displayed no oocyte activation or Ca2+ oscillation-inducing abilities. Although ELS could induce activation at 87, 86 and 31% of injected oocytes respectively, most of the intracellular calcium concentration ([Ca2+]i) responses of ELS-injected oocytes did not show oscillation patterns; only several transient [Ca2+]i rises (transient pattern) were seen. Similarly, with hamster, rabbit and human, while ROS could induce oocyte activation efficiently (70, 71 and 52% respectively), most of the [Ca2+]i patterns of injected oocytes were transient patterns, and not oscillation patterns. When ROS nuclei only from these latter species were injected into mouse oocytes, most of the oocytes could not be activated. [Ca2+]i patterns of oocytes injected with immature sperm cells changed from transient pattern to oscillation pattern while the cells were maturing into spermatozoa. With hamster ROS, oocyte-activating factor was found to be distributed mainly in the cytoplasm. It was interesting that there is a dissociation between the timings of appearance of oocyte activation and that of Ca2+ oscillation of oocytes injected with developing immature sperm cells.

Animals↗
PubMed1989

Distinct mouse DNA sequences enable establishment and persistence of plasmid DNA polymers in mouse cells.

Distinct elements isolated from mouse genomic DNA confer on plasmid DNA the ability to persist at high copy numbers in mouse L fibroblasts (1). Field inversion gel electrophoresis demonstrated that - in contrast to our previous assumption - the persisting plasmid DNA does not exist extrachromosomally but as clusters of tandem repeats integrated into genomic DNA. Digestion with restriction endonucleases that do not cut within the plasmid DNA results in fragments of 50-300 kb in length indicating reiteration of 10-50 plasmid DNA molecules. Restriction with several enzymes that cut once or twice within the plasmid sequences lead to fragment(s) indicative for head-to-tail tandem repeats. In situ hybridization revealed signals for a long homogeneously staining region (HSR) in one or two chromosomes per cell nucleus. Possibilities how these elements could act in the establishment and/or maintenance of the head-to-tail polymers of plasmid DNA in mouse cells are discussed.

Animals↗
PubMed1995

Stimulation of mouse DNA primase-catalyzed oligoribonucleotide synthesis by mouse DNA helicase B.

Many prokaryotic and viral DNA helicases involved in DNA replication stimulate their cognate DNA primase activity. To assess the stimulation of DNA primase activity by mammalian DNA helicases, we analyzed the synthesis of oligoribonucleotides by mouse DNA polymerase alpha-primase complex on single-stranded circular M13 DNA in the presence of mouse DNA helicase B. DNA helicase B was purified by sequential chromatography through eight columns. When the purified DNA helicase B was applied to a Mono Q column, the stimulatory activity for DNA primase-catalyzed oligoribonucleotide synthesis and DNA helicase and DNA-dependent ATPase activities of DNA helicase B were co-eluted from the column. The synthesis of oligoribonucleotides 5-10 nt in length was markedly stimulated by DNA helicase B. The synthesis of longer species of oligoribonucleotides, which were synthesized at a low level in the absence of DNA helicase B, was inhibited by DNA helicase B. The stimulatory effect of DNA helicase B was marked at low template concentrations and little or no effect was observed at high concentrations. The mouse single-stranded DNA binding protein, replication protein A (RP-A), inhibited the primase activity of the DNA polymerase alpha-primase complex and DNA helicase B partially reversed the inhibition caused by RP-A.

Adenosine Triphosphatases↗
PubMed1995

Lens-specific activity of the mouse alpha A-crystallin promoter in the absence of a TATA box: functional and protein binding analysis of the mouse alpha A-crystallin PE1 region.

Lens-specific expression of the mouse alpha A-crystallin gene is regulated at the level of transcription. Here, we have studied the role of the PE1 region, which contains the TATA box (-31/-26) and the immediately adjacent PE1B sequence (-25/-12), in transcriptional regulation. Deletions within either the TATA box or PE1B sequence eliminated promoter activity in transfected lens cells. Surprisingly, these deletions did not eliminate lens-specific promoter activity of the transgene of transgenic mice. Transcription of the transgene with a TATA-deleted promoter initiated at multiple sites in the lenses of the transgenic mice. Footprint analysis revealed that the entire PE1 region was protected by nuclear extracts prepared from lens cells which express the alpha A-crystallin gene and from fibroblasts which do not express the gene. The -37/+3 region formed three specific EMSA complexes using lens cell nuclear extracts, while a similar but much less intense pattern was observed when a fibroblast nuclear extract was used. Competition experiments indicated that these complexes were not due to the binding of TBP to the TATA box, but rather to the binding of other nuclear proteins to the PE1B -25/-19 region. A series of co-transfection competition studies in vivo also suggested the functional importance of proteins binding in the -25/-19 region. The PE1B protein-DNA interactions appear to be conserved in the chicken, rodent and human alpha A-crystallin gene as well as within the alpha A- and alpha B-crystallin genes in the mouse. Our findings indicate that the PE1B region is important for mouse alpha A-crystallin promoter activity; the proximity of this site to the TATA box raises the possibility for cooperativity or competition between TBP and PE1B-bound proteins.

Animals↗
PubMed1997

Two polysialic acid synthases, mouse ST8Sia II and IV, synthesize different degrees of polysialic acids on different substrate glycoproteins in mouse neuroblastoma Neuro2a cells.

We previously cloned cDNAs encoding two different polysialic acid (PSA) synthases, ST8Sia II and IV, from mouse, and showed that both mouse ST8Sia II and IV can synthesize PSA on the neural cell adhesion molecule (NCAM) as well as other glycoproteins such as fetuin, at least in vitro (Kojima, N., Tachida, Y., Yoshida, Y., and Tsuji, S. (1996) J. Biol. Chem. 271, 19457-19463]. In the present study, to clarify how the two PSA synthases act differently in vivo, we first cloned PSA-expressing cell lines (N2a-II and N2a-IV) by stable transfection of the cDNA encoding either mST8Sia II or IV into mouse neuroblastoma Neuro2a cells, which do not express PSA but express NCAM, then compared the expression of the PSA and NCAM isoforms and de novo synthesis of PSA between N2a-II and N2a-IV. Western blotting with an anti-NCAM polyclonal antibody showed that NCAM was expressed as the polysialylated form in both ST8Sia II cDNA-transfected and ST8Sia IV cDNA-transfected Neuro2a cells, but that the polysialylated NCAMs expressed in ST8Sia IV cDNA-transfected clones migrated much slower on SDS-PAGE than those expressed in ST8Sia II cDNA-transfected clones. The slower migration of polysialylated NCAM of the ST8Sia IV cDNA-transfected clone (N2a-IV) than that of the ST8Sia II cDNA-transfected clone (N2a-II) was also observed when cells were metabolically labeled with [3H]glucosamine or pulse-chase labeled with [35S] methionine followed by immunoprecipitation with anti-PSA antibody or anti-NCAM monoclonal antibody. In addition, polysialylated N-glycans of PSA-carrying glycoproteins prepared from [3H] glucosamine-labeled N2a-IV by immunoprecipitation with anti-PSA monoclonal antibody were eluted at a much higher salt concentration than those from [3H] glucosamine-labeled N2a-II on an anion-exchange column. These results indicated that the degree of de novo polysialylation of NCAM by mST8Sia IV was much higher than that by mST8Sia II. In N2a-IV, NCAM-120, -140, and -180 were expressed as polysialylated forms, while polysialylation was restricted to NCAM-140 and -180, i.e., not NCAM-120, in N2a-ST8Sia II. Metabolic labeling of the cells with [3H] glucosamine, pulse-chase labeling with [35S] methionine followed by immunoprecipitation with anti-PSA antibody, and subsequent sialidase treatment revealed that NCAM-140 and -180 were specifically polysialylated in N2a-II, whereas not only NCAM but also other glycoproteins were de novo polysialylated in N2a-IV. The above results demonstrated that the two different PSA synthases, mST8Sia II and IV, synthesize PSA of different lengths on different substrate glycoproteins in vivo when the enzymes are expressed in neuroblastoma Neuro2a cells. These differences suggest that mST8Sia II and IV play different roles in the biosynthesis and expression of PSA.

Animals↗
PubMed2000

Genomic organization and transcriptional regulation of the mouse GD3 synthase gene (ST8Sia I): comparison of genomic organization of the mouse sialyltransferase genes.

The genomic organization of the gene encoding the mouse GD3 synthase (ST8Sia I) has been determined. The mouse ST8Sia I gene spans over 100 kilobases of genomic DNA with a unique genomic structure of 5 exons. Analysis of the sequence immediately upstream of the transcription initiation site revealed that the ST8Sia I promoter contained no canonical TATA- or CCAAT-box, but contained a putative Sp1 binding site. Transient transfection experiments demonstrated functional promoter activity of the ST8Sia I promoter in an ST8Sia I-expressing cell line, P19, but not in an ST8Sia I-nonexpressing cell line, NIH3T3. Mobility shift assay and mutation analysis of the promoter region indicated that the Sp1 binding site is involved in the transcriptional regulation of the ST8Sia I gene in P19 cells. Here, the genomic structural analyses of mouse sialyltransferase genes are summarized and the genomic structures of these genes are compared.

Animals↗
PubMed1983

Inhibitory spectrum of mouse contrapsin and alpha-1-antitrypsin against mouse serine proteases.

Contrapsin and alpha-1-antitrypsin have been recently characterized as major protease inhibitors in mouse plasma (Takahara, H. & Sinohara, H. (1982) J. Biol. Chem. 257, 2438-2446). We have studied the effects of the two inhibitors upon various serine proteases prepared from mouse tissues. Trypsin, plasmin and trypsin-like proteases of the submaxillary gland were inhibited by contrapsin but not by alpha-1-antitrypsin. On the other hand, chymotrypsin, elastase, and thrombin were inactivated by alpha-1-antitrypsin but not by contrapsin. Thus, their inhibitory spectra did not overlap each other in spite of their broad specificities. The inhibition of trypsin, chymotrypsin, and elastase was rapid and stoichiometric, whereas the inhibition of the other proteases was relatively slow. Contrapsin accounted for almost the total capacities of mouse plasma to inhibit both trypsin and submaxillary gland trypsin-like proteases, whereas alpha-1-antitrypsin was responsible for nearly all the capacities of plasma to inhibit both chymotrypsin and elastase.

Animals↗
PubMed2003

Control of mouse hils1 gene expression during spermatogenesis: identification of regulatory element by transgenic mouse.

Histone H1-like protein in spermatids 1 (Hils1) is a testis- specific histone H1-like protein exclusively expressed in haploid spermatids and should be involved in chromatin remodeling during mouse spermatogenesis. Spatial and temporal regulation of the hils1 gene expression would be critical for the formation of functional sperm, controlled at both transcriptional and translational levels. Previously, we reported that transcripts of the hils1 gene are exclusively expressed in mouse testis from 23 days of age whereas the Hils1 protein is not detected until 28 days of age, suggesting that hils1 is a member of a class of translationally regulated genes. By analyzing transgenic mice, we could demonstrate that 318-base pair (bp) 5'-proximal region corresponding to the first 70-bp proximal TATA-less promoter, and 248 bp of 5'-untranslated region is sufficient to confer testis- and spermatid-specific transcription as well as posttranscriptional control of the mouse hils1 gene in vivo.

Animals↗
PubMed1984

Binding of mouse spermatozoa to the zonae pellucidae of mouse eggs in cumulus: evidence that the acrosomes remain substantially intact.

Cumulus-intact mouse eggs inseminated in vitro with capacitated mouse sperm have sperm bound to the zonae pellucidae as early as 20 min postinsemination. Maximal sperm binding was observed at 60 min, after which time the number of sperm bound per egg decreased to about 30% of maximal. This extent of binding remained constant from 120 to 180 min postinsemination. All the sperm bound to the zonae had intact acrosomes as determined by chlortetracycline fluorescence assay. The sperm binding was retarded during the first 60 min by 3-quinuclidinyl benzilate, a specific inhibitor of the acrosome reaction induced by zona glycoprotein, but this compound had no effect on sperm binding during the next 60 min. The observation that sperm, which bind to zonae pellucidae after traversing the cumulus, have intact acrosomes is consistent with the same observation of intact acrosomes made with sperm binding to cumulus-free eggs [Saling and Storey (1979). J. Cell Biol. 83:544-555], and further supports the hypothesis that the sequence of events leading to fertilization of mouse eggs is: binding of intact sperm to the zona pellucida followed by the acrosome reaction induced by zona glycoprotein.

Acrosome↗
PubMed1986

Bicarbonate is essential for fertilization of mouse eggs: mouse sperm require it to undergo the acrosome reaction.

Fertilization of mouse eggs by capacitated mouse sperm occurs readily in vitro in a defined culture medium designated medium CM, derived from Krebs-Ringer bicarbonate medium by supplementation with glucose, lactate, pyruvate, and albumin. Medium CM contains 25 mM HCO-3 equilibrated with CO2 to pH 7.4. Replacement of the CO2/HCO-3 buffer with 25 mM N-hydroxyethylpiperazine-N-ethanesulfonate (HEPES) while otherwise maintaining pH, ionic strength, and composition identical to that of medium CM, yielded medium HM in which the percentage of in vitro fertilization observed was consistently zero. Addition of 10 mM HCO-3 to medium HM yielded medium HMB, in which the percentage of eggs fertilized was 70%, equal to that observed in medium CM. This percentage applies to cumulus-free, zona-intact eggs. With zona-free eggs, the percentage fertilization increased to 90% in media CM and HMB, but remained zero in medium HM. This result shows that HEPES does not inhibit fertilization while omission of CO2/HCO-3 blocks it completely. Capacitation of mouse sperm, as determined by chlortetracycline fluorescence assay, proceeded at the same rate in media CM and HMB, but was retarded in medium HM. Sufficient sperm became capacitated, however, that this could not be the block of fertilization. The acrosome reaction in sperm bound to isolated zonae pellucidae, as also determined by chlortetracycline fluorescence assay, occurred with the same time course in media CM and HMB, but did not occur in medium HM.(ABSTRACT TRUNCATED AT 250 WORDS)

Acrosome↗
PubMed2004

Differential expression of the trypsin inhibitor SPINK3 mRNA and the mouse ortholog of secretory granule protein ZG-16p mRNA in the mouse pancreas after repetitive injury.

A mouse model using repetitive acinar cell injury caused by supraphysiologic doses of cerulein to induce the characteristic fibrosis and loss of acinar cell mass found in human chronic pancreatitis was employed to identify early changes in gene expression. A gene array was used to detect changes in 18,000 expressed sequence tags; changes in specific transcripts were confirmed by RNase protection assays. These methods identified SPINK3, the mouse homologue of human and rat protease inhibitor genes, as being highly expressed in the pancreas and induced after pancreatic injury. Because SPINK3 may be an important serine protease inhibitor, its up-regulation may reflect an important endogenous cytoprotective mechanism in preventing further injury. The up-regulation of SPINK3 was specific; the mouse homologue of the zymogen-processing protein ZG-16p was also highly expressed in the pancreas but sharply down-regulated early in the course of injury. These findings suggest that the pancreatic acinar cell may respond to injury with a program of self-preservation and loss of normal function.

Amino Acid Sequence↗
PubMed2000

Immunobiology of xenogeneic cornea grafts in mouse eyes. II. Immunogenicity of xenogeneic cornea tissue grafts implanted in anterior chamber of mouse eyes.

BACKGROUND: Xenogeneic corneal fragments (guinea pig) are highly resistant to immune rejection in the anterior chamber of mouse eyes. Because guinea pigs and mice are discordant (i.e., mouse serum normally contains guinea pig reactive xenoantibodies), we wished to determine the extent to which xenogeneic corneal fragments placed intraocularly in normal and specifically sensitized mice activated xenoreactive T and B cells. METHODS: Guinea pig corneas, deprived surgically of epithelium, were cut into fragments and inserted into the anterior chamber of eyes of BALB/c mice, adjacent to the central cornea of the recipient. Antibody (immunoglobulin [Ig]M, IgG) and delayed type hypersensitivity (DTH) immune responses of recipient mice to guinea pig xenoantigens were assessed. The fate of xenogeneic cornea implants was assessed in mice immunized systemically to guinea pig antigens. RESULTS: Guinea pig spleen cells and corneal fragments implanted s.c. induced within 2 weeks of immunization both DTH and IgG antibodies to guinea pig xenoantigens. By contrast, xenogeneic corneal fragments implanted in the anterior chamber of mouse eyes evoked no change in recipient humoral immune status and induced mild guinea pig-specific DTH only after 5 weeks. Presensitization of mice to guinea pig xenoantigens failed to increase the proportion of grafts that were regarded as rejected, but the onset of rejection in failed grafts occurred earlier than in unsensitized recipients. Active systemic immunization of mice bearing intracameral guinea pig corneal fragments failed to curtail the grafts' survival. Guinea pig corneal fragments implanted in the anterior chamber of normal mice failed to induce anterior chamber-associated immune deviation. CONCLUSIONS: Xenogeneic corneal fragments implanted in the anterior chamber of eyes of normal mice display strikingly reduced immunogenicity, and an inability to induce anterior chamber-associated immune deviation. These properties are discussed in terms of the vulnerability of guinea pig corneal tissue to immune rejection within the eye.

Animals↗
PubMed2002

The eyes of mito-mouse: mouse models of mitochondrial disease.

The recent creation of several mouse models of mitochondrial diseases has provided new insights into the understanding of human mitochondrial disorders. Whether these animals have clinical or histologic ophthalmologic abnormalities is of great interest given the high frequency of such abnormalities in humans with mitochondrial disorders. In this article, we describe the currently available mouse models for mitochondrial diseases with special emphasis on their ocular phenotype. These mouse models demonstrate multiple and varied ophthalmologic manifestations.

Animals↗
PubMed1982

Interferon production in mouse spleen cells and mouse fibroblasts (L cells) stimulated by various strains of Newcastle disease virus.

Interferon production in mouse spleen cells and mouse fibroblasts (L cells) stimulated by three strains of Newcastle disease virus (NDV), Italian, La Sota and Ulster, was investigated. Strain Italian was fully infectious and highly virulent; strain Ulster exhibited very low infectivity and very low virulence; strain La Sota was between these extremes. All of these strains of MDBK cell-grown NDV could induce interferon in mouse spleen cells, and it was concluded that proteolytic cleavage of F0 protein of NDV and, consequently, virus penetration are not necessary for interferon induction in these cells. On the other hand, NDV with uncleaved F0, which was characterized by an apparent lack of haemolytic and cell fusion activity and infectivity for tissue culture cells, had no interferon-inducing ability in L cells. The cleavage of F0 protein was paralleled by an appearance of interferon-inducing activity in L cells.

Animals↗
PubMed1984

In vitro mouse cytomegalovirus infection of mouse tracheal epithelial cells requires the presence of other cell types.

Recently, methods have been developed to culture dissociated tracheal epithelial (TE) cells. Attempts were made to infect these epithelial cells with mouse cytomegalovirus (MCMV) to see if the dissociated epithelial cells share characteristics of infection with MCMV-infected tracheal organ cultures. When isolated TE cells were incubated with MCMV at multiplicities between 0.10 and 2.0, no infection or minimal infection resulted. Centrifugation of virus onto the TE cell sheets also resulted in only minimal infection. If MCMV-infected mouse embryo fibroblasts were added to cultures of TE cells, however, the TE cells subsequently became infected. TE cells could also be infected by incubating MCMV with mixed cultures of uninfected fibroblasts and TE cells. The epithelial nature of infected cells was confirmed by electron microscopy. Reconstruction experiments demonstrated that fibroblast-mediated infection of mouse TE cells with MCMV was not simply due to the large amounts of virus provided by the infected fibroblasts. It is suggested that cell-cell contact, or fusion with infected cells is required for productive infection of TE cells with MCMV.

Animals↗
PubMed2002

Analysis of 16S libraries of mouse gastrointestinal microflora reveals a large new group of mouse intestinal bacteria.

Total genomic DNA from samples of intact mouse small intestine, large intestine, caecum and faeces was used as template for PCR amplification of 16S rRNA gene sequences with conserved bacterial primers. Phylogenetic analysis of the amplification products revealed 40 unique 16S rDNA sequences. Of these sequences, 25% (10/40) corresponded to described intestinal organisms of the mouse, including Lactobacillus spp., Helicobacter spp., segmented filamentous bacteria and members of the altered Schaedler flora (ASF360, ASF361, ASF502 and ASF519); 75% (30/40) represented novel sequences. A large number (11/40) of the novel sequences revealed a new operational taxonomic unit (OTU) belonging to the Cytophaga-Flavobacter-Bacteroides phylum, which the authors named 'mouse intestinal bacteria'. 16S rRNA probes were developed for this new OTU. Upon analysis of the novel sequences, eight were found to cluster within the Eubacterium rectale-Clostridium coccoides group and three clustered within the Bacteroides group. One of the novel sequences was distantly related to Verrucomicrobium spinosum and one was distantly related to Bacillus mycoides. Oligonucleotide probes specific for the 16S rRNA of these novel clones were generated. Using a combination of four previously described and four newly designed probes, approximately 80% of bacteria recovered from the murine large intestine and 71% of bacteria recovered from the murine caecum could be identified by fluorescence in situ hybridization (FISH).

Animals↗
PubMed2000

The mouse brain transcriptome by SAGE: differences in gene expression between P30 brains of the partial trisomy 16 mouse model of Down syndrome (Ts65Dn) and normals.

Trisomy 21, or Down syndrome (DS), is the most common genetic cause of mental retardation. Changes in the neuropathology, neurochemistry, neurophysiology, and neuropharmacology of DS patients' brains indicate that there is probably abnormal development and maintenance of central nervous system structure and function. The segmental trisomy mouse (Ts65Dn) is a model of DS that shows analogous neurobehavioral defects. We have studied the global gene expression profiles of normal and Ts65Dn male and normal female mice brains (P30) using the serial analysis of gene expression (SAGE) technique. From the combined sample we collected a total of 152,791 RNA tags and observed 45,856 unique tags in the mouse brain transcriptome. There are 14 ribosomal protein genes (nine under expressed) among the 330 statistically significant differences between normal male and Ts65Dn male brains, which possibly implies abnormal ribosomal biogenesis in the development and maintenance of DS phenotypes. This study contributes to the establishment of a mouse brain transcriptome and provides the first overall analysis of the differences in gene expression in aneuploid versus normal mammalian brain cells.

Animals↗
PubMed2004

Organization and evolution of a gene-rich region of the mouse genome: a 12.7-Mb region deleted in the Del(13)Svea36H mouse.

Del(13)Svea36H (Del36H) is a deletion of approximately 20% of mouse chromosome 13 showing conserved synteny with human chromosome 6p22.1-6p22.3/6p25. The human region is lost in some deletion syndromes and is the site of several disease loci. Heterozygous Del36H mice show numerous phenotypes and may model aspects of human genetic disease. We describe 12.7 Mb of finished, annotated sequence from Del36H. Del36H has a higher gene density than the draft mouse genome, reflecting high local densities of three gene families (vomeronasal receptors, serpins, and prolactins) which are greatly expanded relative to human. Transposable elements are concentrated near these gene families. We therefore suggest that their neighborhoods are gene factories, regions of frequent recombination in which gene duplication is more frequent. The gene families show different proportions of pseudogenes, likely reflecting different strengths of purifying selection and/or gene conversion. They are also associated with relatively low simple sequence concentrations, which vary across the region with a periodicity of approximately 5 Mb. Del36H contains numerous evolutionarily conserved regions (ECRs). Many lie in noncoding regions, are detectable in species as distant as Ciona intestinalis, and therefore are candidate regulatory sequences. This analysis will facilitate functional genomic analysis of Del36H and provides insights into mouse genome evolution.

Animals↗