Major Organs Of Mice Research Articles

Effect of lethal <I>Vibrio vulnificus</I> infection on blood system and pathology changes of major organs in mice

Effect of lethal <I>Vibrio vulnificus</I> infection on blood system and pathology changes of major organs in mice

Conserved expression of the PRELI domain containing 2 gene (Prelid2) during mid-later-gestation mouse embryogenesis

Prelid2, which belongs to the PRELI domain containing family, is identified as a conserved evolution gene. The expression and regulation during embryonic development of the prelid2 gene is unknown. In this study, we investigated the prelid2 gene expression and regulation using mouse embryos model, by in situ hybridization analysis, RT-PCR and bisulfite sequencing. In situ hybridization analysis showed that prelid2 gene expression were found in midbrain, spinal cord, optic eminence, otic vesicle and tail at E9.5 and E10.5 embryos, in forebrain, hindbrain, heart, lung, liver and kidney at E13.5 and E15.5 embryos. Real-time quantitative RT-PCR results verified the expression pattern in the four major mouse organs, brain, heart, lung, and liver during organs differentiation and formation. Bisulfite sequencing illustrated the consistent result of expression and its unmethylation status in the genomic promoter region at E12.5, E18.5, and new born. Thus, the prelid2 gene is a widely-spread, persistently expressed and unmethylated gene in mouse embryonic development. Our results suggest that the PRELI domain containing 2 gene is involved in mouse embryonic development.

A novel mouse protein differentially regulated by androgens in the submandibular and lacrimal glands

We characterized a cDNA clone derived from the female mouse submandibular gland (SMG). The transcript of this cDNA was approximately 1.2 kb in size and predicted to code a 165-amino acid protein with a putative signal peptide for a secretory pathway. This protein, named submandibular androgen-repressed protein (SMARP), had homology in the N-terminal region with members of the glutamine/glutamic acid-rich protein (GRP) family from rats. Northern blot analysis revealed that SMARP mRNA is expressed, out of the major mouse organs, only in the SMG and exorbital lacrimal gland (LG), with much more abundance in the former. For the SMG, the level of SMARP mRNA was 36 times higher in females than males, whereas for the LG it was 28 times higher in males than females. Furthermore, the level of SMARP mRNA was increased in the SMG but reduced in the LG with castration in males, whereas it was reduced in SMG but increased in LG after administration of testosterone in females or castrated males. In situ hybridization detected the signal for SMARP mRNA in the female SMG, and immunohistochemistry detected the signal for SMARP protein in the female SMG and male LG. In the female SMG, SMARP mRNA, and protein were localized intensively in a subpopulation of acinar cells, whereas in the male LG, SMARP protein was distributed diffusely in all acinar cells. These results suggested that SMARP is a secretory protein whose expression is regulated by androgens negatively in the SMG and positively in the LG.

Evaluation of tumor-specific promoter activities in melanoma

Gene therapy is a novel therapy for melanoma. To date, however, there is still no powerful tumor specific promoter (TSP) to restrict the transgene expression in melanoma cells. In order to define a useful TSP for targeting in the context of melanoma gene therapy, four promoters, the cyclooxygenase-2 (Cox-2), alpha-chemokine SDF-1 receptor (CXCR4), epithelial glycoprotein 2 (EGP-2), and survivin, were tested in both established melanoma cell lines and primary melanoma cells. We employed recombinant adenoviral vectors (reAds) each with a candidate TSP (the Cox-2, CXCR4, EGP-2, or survivin), a reporter luciferase gene, and a poly-A signal, all of which were inserted into the E1-deleted region. A reAdGL3Bcytomegalovirus (CMV), containing the CMV promoter and luciferase gene, was used as a positive control to normalize the luciferase activity. Luciferase activity was measured in multiple tumor cell lines and two primary melanoma cell cultures after infection with reAds. Human epithelial melanocytes, HEM, were used as normal control. In contrast to three other promoters, the survivin promoter exhibited the highest activities within both melanoma cell lines and primary melanoma cells, but not in HEMs. Additionally, the survivin promoter exhibited very low activities in major mouse organs including the liver, in vivo. EGP-2 is not active in melanoma; messenger RNA expressions were correlated to promoter activities both in melanoma cell lines and primary cell cultures. Thus, these data suggest that the survivin promoter achieved a 'tumor-on/liver-off' profile, and thus represents a potentially useful tumor-specific promoter with applications for transcriptional targeting of Ad vector-based cancer gene therapy or oncolysis to melanoma.

SUB-LETHAL INJECTION OF COBRA VENOM DECREASES ADENOSINE DEAMINASE, HISTAMINE, AND IgE IN ORGANS OF MICE

The pharmacological substances adenosine deaminase (ADA), histamine, and IgE are endogenously present in animals. They are implicated in allergy and asthma and detectable in blood serum. This research reports the presence of ADA, histamine, and IgE at varying levels in almost all major organs of mice. This research further reports that intramuscular injection of sub-lethal dose of Naja kaouthia venom disrupted homeostasis and lowered the levels of ADA, histamine, and IgE in the organs of mice. Adult Balb/C male mice were injected with a half lethal dose of the venom. The mice were sacrificed at 2, 8, and 24 hours post-injection and different organs were collected. Organs were homogenized, centrifuged, and the supernatants were assayed for ADA, histamine, and IgE using respective antisera by immunological test enzyme-linked immunosorbent assay (ELISA). Organs from mice injected with PBS served as controls. No major decrease in the levels of ADA, histamine, and IgE was observed after 2 hours of venom injection. However, tremendous decreases in the levels of ADA, histamine, and IgE was observed in organs 24 h post-injection. The highest decrease for ADA was observed in the brain, liver, lung, muscle, and testis; for histamine, in the heart, muscle, lung, and testis; and for IgE, in the bone, heart, lung, muscle, and testis. This is a first-hand investigation showing the effect of envenomation on the pharmacokinetics of ADA, histamine, and IgE in organs.

Lesions of aryl-hydrocarbon receptor-deficient mice.

We have analyzed the possible role of the aryl-hydrocarbon receptor (AHR) in the aging process of mice using a homozygous null mouse (Ahr-/-) line as a model. We studied 52 male and female Ahr-/- mice aged from 6-13 months. Forty-six percent died or were ill by 13 months of age. Ahr-/- mice developed age-related lesions in several organs, some of which were apparent after only 9 months of age. Cardiovascular alterations included cardiomyopathy (100%) with hypertrophy and focal fibrosis. Vascular hypertrophy and mild fibrosis were found in the portal areas of the liver (81%), and vascular hypertrophy and mineralization were common in the uterus (70%). Gastric hyperplasia that progressed with age into polyps was evident in the pylorus of 71% of the mice over 9 months of age. Ahr-/- mice had T-cell deficiency in their spleens but not in other lymphoid organs. The immune system deficiency described previously could be the origin for the rectal prolapse found in 48% of the null mice, associated with Helicobacter hepaticus infection. In the dorsal skin (53% incidence), severe, localized, interfollicular and follicular epidermal hyperplasia, with hyperkeratosis and acanthosis, and marked dermal fibrosis, associated with the presence of anagenic hair follicles, were also evident. None of these lesions were found in 42 control (Ahr +/+ or +/-) mice of similar ages. These observations suggest that the AHR protein, in the absence of an apparent exogenous (xenobiotic) ligand, plays an important role in physiology and homeostasis in major organs in mice, and further supports an evolutionary conserved role for this transcription factor.

Isolation of a mouse cDNA encoding mSTI1, a stress-inducible protein containing the TPR motif

We report the isolation and sequencing of the complete 2079-bp cDNA fragment encoding mSTI1, a murine stress-inducible protein. The predicted ORF encodes a protein of 543 amino acids (aa) and M r 62 582. The predicted protein has significant homology to stress-inducible proteins from humans (IEF SSP 3521), soybean (GMSTI), yeast (STI1) and a parasite, Leishmania donovani (LSIP). All of these proteins contain 34-aa repeat motifs, termed tetratricopeptide repeats (TPRs), that are proposed to be involved in intra- and intermolecular protein interactions. mSTI1 has ten potential TPR motifs, a putative nuclear localization signal (NLS), six potential phosphorylation sites for casein kinase II and a central proline-rich region. Western analysis detected a protein of approx. 63 kDa in all the major mouse organs and in mouse, monkey and human cell lines.

Biodistribution of synthetic thymosin α1 in the serum, urine and major organs of mice

Thymosin fraction 5 (TF5), a thymic preparation, has been shown to be an immune-potentiating agent consisting of biologically active polypeptide components with hormone-like activities. Thymosin α 1 ( Tα 1) was the first biologically active polypeptide to be purified from TF5 and completely characterized. It is an acidic peptide with an isoelectric point of 4.2 and a molecular weight of 3108. Tα 1 is considered a biological response modifier which amplifies T-cell immunity. In the present study, we have studied some pharmacokinetic properties of Tα 1 by measuring its concentrations in serum, urine and ten major organs of female Swiss-Webster mice following administration of 500 μg Tα 1 intraperitoneally. Using a modified enzymatic immunoassay, our data show a significant increase of Tα 1 in serum 2min after injection and lasting for 2 h (average: 1.55 ± 0.27 μg/ml). In urine, at four different time points after injection (20min, 40 min, 2h, 6 h), increased concentrations of Tα 1 were found between 24.2 and 25.4μg/ml (average: 25 ± 0.47 μg/ml). Of the 500μg Tα 1 administered to mice, 8.97% was recovered at the end of the study, of which 2% corresponded to urine, 1.25% to serum (2 ml of serum per mouse), and 5.72% to organs. Since the urine/day volume and the serum volume of any Swiss-Webster mouse is ca 2 ml, additional extrapolation of the above mentioned values could show percentages of recovery close to 40% for urine and 2.5% for serum. In most of the organs, the wet weight concentrations of Tα 1 increased significantly during the first 40 min after injection in comparison to their baseline wet weight concentrations. These organs consisted of the following: thymus (33.1 ± 3.5 μ/g vs 18 μ/g baseline); lungs (7.7±1.1 μg/g vs 1.9 μg/g baseline); spleen (15.6±0.7 μg/g vs 5.6 μg/g); kidneys (6.2±1.1 μg/g vs 3.9 μg/g); ovaries (9.2±1.4 μg/g vs 0 μg/g); and peritoneal fat (4±1 μg/g vs 0 μg/g). No significant increases were observed in the liver (1.7±0.1 μg/g vs 1.4 μg/g) and heart (0.7±0.5 μg/g vs 0 μg/g). Increased concentrations of Tα 1 were not detected in the brain and skeletal muscle tissues. These pharmacokinetic studies of Tα 1 in mice indicate that rapid renal excretion of Tα 1 represents a major source of humoral loss following IP administration. Recent preliminary studies in humans confirm that the kidney rapidly releases high levels of Tα 1 in urine in a time frame consistent with that observed in mice.

Biodistribution of synthetic thymosin β4 in the serum, urine, and major organs of mice

Thymosin β 4 (T β 4) is a peptide of 43 amino acids that was first isolated from the thymus gland and subsequently found to be ubiquitous in nature. T β 4 functions mainly as an actin-sequestering molecule in non-muscle cells, where its primary role is to maintain the large pool of unpolymerized G-actin in the cell. Studies on the pharmacokinetics of T β 4 in human and other mammals have not been reported so far. In the present study, we have measured T β 4 concentrations in serum, urine, and 10 major organs of female Swiss-Webster mice following intraperitoneal administration of 400 μg synthetic T β 4. Using a modified enzymatic immu-noassay, our data show a significant increase of T β 4 in serum starting 2 min after injection and lasting for 40 min (average: 2.34±0.54μg/ml). High concentrations were found in urine (59.3 ± 7.54μg/ml) at three different time points after injection (20 min, 40 min, and 2 h). Of the 400 μg T β 4 administered to mice 83 % was recovered at the end of the study, 44.6% of which corresponded to urine, 1.4% to serum, and 37.5% to the organs. In 50% of the tested organs, the wet weight concentrations of T β 4 increased significantly from the first 40 min to 2 h after injection in comparison to their baseline wet weight concentrations. These organs were: the brain (72 μg/g vs 42 μg/g), heart (80 μg/g vs 37 μg/g), liver (15 μg/g vs 9 μg/g), kidneys (65 μg/g vs 28 μg/g), and peritoneal fat (47 μg/g vs 13 μg/g). Wet weight concentrations increased in the thymus (196 μg/g vs 147 μg/g) and muscle (45 μg/g vs 0 μg/g) after 6 h of injection. The spleen showed an increase in wet weight concentrations at the 2 min timepoint (267 μg/g vs 161 μg/g). Ovaries had a biphasic increase at 40 min(72 μg/g vs 62 μg/g) and 24 h (92 μg/g vs 62 μg/g) after T β 4 administration. In lungs, the highest wet weight increase after injection (149 μg/g at timepoint 6h) was not higher than its basal wet weight concentration (153 μg/g). These phar-macokinetic studies of T β 4 in mice have established that high levels of T β 4 are found in the blood following I.P. administration and the kidney rapidly removes the peptide from the circulation. The kinetics of this response should help define the proper scheduling of administration of T β 4 during clinical trials in disorders, such as the acute respiratory distress syndrome (ARDS), associated with actin toxicity.

Natural mouse IgG reacts with self antigens including molecules involved in the immune response.

IgG isolated on protein A-Sepharose from pools of normal sera from various mouse strains were examined by immunoblotting for reaction with self antigens. Homogenates of the major mouse organs, i.e. brain, skin, spleen, kidney, adrenals, thymus, heart, muscle and liver were used as the source of autoantigens. IgG stained at least 220 bands on the immunoblots. The antigens corresponding to these bands were tentatively identified by molecular mass estimation and referenced to computerized mouse protein data banks. IgG mainly recognized enzymes but it also stained intracellular structural constituents and surface molecules implicated in the functioning of the immune system. The validity of this identification was confirmed by analyzing purified antigens from mouse or other animal species by immunoblotting and enzyme immunoassays. Furthermore, extracts of 125I-surface-labeled cells were immunoprecipitated with IgG in the liquid phase or immobilized on beads. The proteins precipitated migrated to the same positions as those precipitated by specific monoclonal antibodies (mAb), such as class I alpha chain and beta 2-microglobulin, class II alpha and beta chains, CD3, CD4 and CD8 antigens. The results obtained with several enzyme immunoassay procedures using cell membrane extracts, specific mAb and normal IgG further supported the specific interaction of IgG with Ia, CD4 and CD8 molecules. Affinity chromatography indicated that at least 20% of normal mouse IgG possess polyreactive autoantibody function. Dissociation constants of these IgG were calculated for some autoantigens and found to be in the range of 2 x 10(-6)-7 x 10(-6) M. It is concluded that normal mouse IgG exhibit autoreactivities similar to those previously described for IgM.

Labeling and comparative biodistribution of the monoclonal antibody [formula omitted] in nude mice bearing human lung adenocarcinoma

In order to evaluate some of the key factors that may allow the optimization of radiolabeled monoclonal antibodies for use as diagnostic and therapeutic tools to detect and treat human neoplasia, we compared the biodistribution of the anti-lung-tumor monoclonal antibody KS 1 4 , labeled with four different radionuclides, in athymic (nu/nu) mice bearing human lung adenocarcinoma. Several radiolabeling methods were used: the first involved coupling a suitable bifunctional chelating agent, such as DTPA, to the KS 1 4 monoclonal antibody, followed by binding the radiometal, either 113mIn or 111In. Radioiodination was carried out by the chloramine-T method with 131I, and intrinsic labeling by generating the hybridoma in the presence of 75Se-methionine. An examination of tumors and major organs of mice injected with one of the above radiolabeled KS 1 4 MoAbs, and biodistribution at various time intervals up to 96 h post injection revealed that iodination and intrinsic labeling yield the highest tumor uptake. Because of the relatively high deiodination that occurs in vivo, and the high 75Se content in the circulation, the preferential uptake (tumor-to-blood ratio) of these radiopharmaceuticals lags behind the equivalent ratios for the In-labeled MoAb. In the latter group of animals, a second consecutive injection of the labeled MoAb resulted in elevated blood level of radioindium, as well as a corresponding decrease in tumor-to-blood ratios.