• The Korean Journal of Zoology
• /
• v.39 no.2
• /
• pp.215-222
• /
• 1996

In the previous studies, we have demonstrated that prorenin converting enzyme (PRECE) was identical to the epidermal grouch factor-binding protein (EGF-BP) type B, which was a member of the mouse glandular kallikrein family, To examine whether or not EGF-BP type A and C are involved in the processing of prorenin, we have cloned the CDNAS of the EGF-BP type h and C from a library of male ICR mouse submandibular gland (SMGI. And then CHO cells were transfected with the EGF-BP expression plasmids. and stable cell lines expressing a high level of the EGF-BPS precursor were obtained. The conditioned medium was then treated with trypsin, which has been knotvn to effectively convert the EGF-BP type A and C precursor to the active forms. 수ubsequentlv, the prorenin converting activity of the trypsin-treated or untreated medium was examined. PRECE converted exactly prorenin to renin, but the prorenin converting activities of EGF-BP type A and C were not detected. From these results, it seems that only type B of these EGF-BPs is involved in processing Ren 2 prorenin in mouse SMG.

• Mass Spectrometry Letters
• /
• v.12 no.3
• /
• pp.112-117
• /
• 2021

α-Amanitin and β-amanitin are highly toxic bicyclic octapeptides responsible for the poisoning of poisonous mushrooms such as Amanita, Galerina, and Lepiota by inhibiting RNA polymerase II, DNA transcription, and protein synthesis. A sensitive, simple, and selective liquid chromatography-high resolution mass spectrometric method using parallel reaction monitoring mode was developed and validated for the simultaneous determination of α- and β-amanitin in mouse plasma to evaluate the toxicokinetics of α- and β-amanitin in mice. Protein precipitation of 5 μL mouse plasma sample with methanol as sample clean-up procedure and use of negative electrospray ionization resulted in better sensitivity and less matrix effect. The calibration curves for α- and β-amanitin in mouse plasma were linear over the range of 0.5-500 ng/mL. The intra- and inter-day coefficient of variations and accuracies for α- and β-amanitin at four quality control concentrations were 3.1-14.6% and 92.5-115.0%, respectively. The present method was successfully applied to the toxicokinetic study of α- and β-amanitin after an oral administration of α- and β-amanitin at 1.5 mg/kg dose to male ICR mice.

• Proceedings of the Zoological Society Korea Conference
• /
• 1999.10b
• /
• pp.31-31
• /
• 1999

Characterization of mutant mice has been utilized as an animal model for the study of human inherited diseases. In addition to the pathogenesis stduy using the mutant mice, the mice have been used for the identification of the genes causing the phenotypes. Functional cloning and positional cloning are two approaches, depending on the phenotypes of the mutant mice. Though it takes a long time positional cloning has been well used to identify the gene of which function can not be presumed from the mouse phenotype. Recently by the advance of the molecular tools and the human genome project close to 10,000 genetic markers are developed to make the procedure faster. We obtained a new mutant mouse, sims, spontaneously arose and the affected mouse has a mild tremor and seizure was observed. Homozygote in either sex is sterile since uterus growth in female and seminal vesicle in male are not induced for the growth in puberty, implying the abnormal hormonal regulation during puberty. Supporting this, there is no detectable testosterone in the serum of the mutant male and the brain of the mutant is 30% heavier than littermate. To identify the location of the mutated gene, intraspecies cross to CAST/Ei was carried out and the 37 affected mice was analyzed for the linkage. The gene was mapped on chromosome 18, 20 cM from the centromere. More than 500 F2 progenies have been analyzed for the linkage and the locus becomes narrow within 3cM between Egrl and Fgf gene.f gene.

• Applied Microscopy
• /
• v.39 no.1
• /
• pp.17-25
• /
• 2009

This study investigated that exposure of male mice to estrogen receptor $\alpha$-selective agonist, 4,4',4"-(4-propyl-[1H]-pyrazole-1,3,5-triyl)tris-phenol (PPT) induce morphological changes of accessory genital glands. The male reproductive organs were fixed and processed for light microscopy. The PPT induced decreases of ventral prostates, seminal vesicles and preputial glands weights with experimental time. The glandular lumen of ventral prostate was atrophied compared with control group. Type of epithelial tissues in the prostate was changed from simple columnar epithelium to stratified cuboidal or squamous epithelium. Treated group with the agonist showed that increased connective tissue underlying epithelium in the prostate and seminal vesicle. Especially, the glandular lumen of the seminal vesicle was contracted when PPT-treated animals were compared with control group. Secretion cells of preputial gland were smaller than that of control group. On week 8, PPT treatment caused decrease of epithelial cell height lining the lumen of preputial gland. These results provide information useful in researching the physiological function of estrogen mediated by estrogen receptor $\alpha$ in male accessory genital gland.

• Animal cells and systems
• /
• v.10 no.4
• /
• pp.227-231
• /
• 2006

Micro-deletions at specific loci of the Y chromosome have been observed frequently in male infertility patients, suggesting that genes in these regions are involved in male germ cell development. DAZ is a representative male infertility gene at the AZFc locus of the Y chromosome. Since DAZ contains an RNA binding motif along with so-called a DAZ domain, it was proposed to participate in RNA metabolism during spermatogenesis. A mouse gene homologous to the human DAZ gene has been cloned and named Dazl (DAZlike). Dazl is autosomal and expressed in the testis and also at a low level in the ovary. Male mice homozygous for the Dazl null allele have small testes with a few spermatogonia and almost complete absence of germ cells beyond the spermatogonial stage, suggesting the requirement of Dazl for entry or progression through meiosis. However, its exact cellular functions have not been understood yet. In order to investigate cellular functions of Dazl, we decided to isolate candidate interacting protein genes of the mouse Dazl, using yeast two-hybrid screening. A number of candidate Dazlinteracting proteins have been isolated, such as Bprp, Acf, Hgs, Murr1, Nbak3 and Ranbp9, but dynein light chain 1 (Dlc1) was most predominant. A strong interaction of Dazl with Dlc1 suggests that Dazl might function as an mRNA adaptor to the dynein motor complex.

• Journal of Embryo Transfer
• /
• v.30 no.3
• /
• pp.213-218
• /
• 2015

In the study for a differentiation and development of spermatogonial cells, the researchers should commonly require a simple, fast and reasonable method that could evaluate the developmental stage of male germ cells without any damage and also relentlessly culture them so far as a cell stage aiming at experimental applications. For developing the efficient method to identify the stage of sperm cells, the morphological characteristics of sperm cells were investigated by staining the cells with blue fluorescent dye Hoechst 33258, and a criterion for male germ cell classification was elicited from results of the previous investigation, then the efficiency of the criterion was verified by applying it to assort the germ cells recovered from male mice in age from 6 to 35 days. As morphological characteristics, spermatogonia significantly differed from spermatocytes in size, appearance and fluorescent patches of nucleus, and spermatids could also be distinguished from spermatozoa by making a difference in the volume and shape of nucleus and the shape and fluorescence of tail. Aforesaid criterion was applicable for classifying in vitro cultured sperm cells by verifying its efficiency and propriety for assorting the stages of testicular germ cells. However, the fluorescent staining showed that germ cells in mouse testis should be dramatically differentiated and developed at 21 days and 35 days of age, which were known as times of sexual puberty and maturity in male mice, respectively. In conclusion, the results indicated that this simple criterion for sperm cell classification using fluorescence staining with Hoechst 33258 may be highly efficient and reasonable for spermatogenesis study.

• Journal of mucopolysaccharidosis and rare diseases
• /
• v.4 no.1
• /
• pp.26-36
• /
• 2018

Mucopolysaccharidosis IIIA is a heritable neurodegenerative disorder resulting from the dysfunction of the lysosomal hydrolase sulphamidase. This leads to the primary accumulation of the complex carbohydrate heparan sulphate in a wide range of tissues and CNS degeneration. Characterization of animal model is the beginning point of the therapeutic clinical trial. Mouse model has a limitation in that it is not a human and does not have all of the disease phenotypes. Therefore, delineate of the phenotypic characteristics of MPS IIIA mouse model prerequisite for the enzyme replace treatment for the diseases. We designed 6-month duration of phenotypic characterization of MPS IIIA mouse biochemically, behaviorally and histologically. We compared height and weight of MPS IIIA mouse with wild type from 4 weeks to 6 months in both male and female. At 6 months, we measured GAG storage in urine kidney, heart, liver, lung and spleen. The brain GAG storage is presented with Alcian blue staining, immunohistochemistry, and electron-microscopy. The neurologic phenotype is evaluated by brain MRI and behavioral study including open field test, fear conditioning, T-maze test and Y-maze test. Especially behavioral tests were done serially at 4month and 6month. This study will show the result of the MPS IIIA mouse model phenotypic characterization. The MPS IIIA mouse provides an excellent model for evaluating pathogenic mechanisms of disease and for testing treatment strategies, including enzyme or cell replacement and gene therapy.

• Parasites, Hosts and Diseases
• /
• v.44 no.2 s.138
• /
• pp.167-169
• /
• 2006

After infection of male mice with the plerocercoids (spargana) of Spirometra mansoni, serum levels of estrogen and testicular weight were analyzed by enzyme-linked immunosorbent assay (ELISA) and weighing machine, respectively. The serum level of estrogen increased progressively in infected mice compared with normal controls, whereas the testicular weight of infected mice decreased significantly (P < 0.05). These results suggest that certain substances from spargana change the steroid hormone metabolisms in the host by unknown pathways, and chronic infection may contribute to change of the function of steroid hormone target organ, i.e., testis, in male mice.

• Journal of Environmental Health Sciences
• /
• v.15 no.2
• /
• pp.75-83
• /
• 1989

To evaluate the effect of Kam Doo-extract (KDE) on organophosphorous (OP)intoxication in mouse, this research was conducted. KDE prescribed with the equal weights of both Padix Glycyrrizae and Simen Glycine was extracted in water at 100$^{\circ}$C for 2hr, and concentrated in a vacuum evaporator. Animmal used in this research was ICR-strained male mice (bodyweight: 20 ~ 25g), and induction material for OP intoxication was DDVP(Dichlovos). Toxicity parameters used to evaluate KDE-preventive effect on DDVP were cholinesterase activity, and protection index of KDE against LDso values of DDVP. As the results, KDE prevented the inhibition of cholinesteranse activity due to DDVP-treatment, and inhanced the protecion index. Consequently our experimental data show the KDE will be useful as an preventive agent in respect that KDE is safe and effective against OP intoxication in mouse.

• Korean Journal of Animal Reproduction
• /
• v.16 no.4
• /
• pp.335-340
• /
• 1993

The development of efficient method for the production of transgenic mice has been investigated in our laboratory. This study was conducted to develop the artificial insemination in the mouse. Spermatozoa were collected from the cauda epididymis of ICR males(age:12~15 weeks, Body weight : 30g) and artificially inseminated into the intrauterine via cervix of hormone-primed ICR females(age: 6~8 weeks, body weight: 25g) using the capillary tube, 200~300 $\mu$m in inner diameter. The effect of concentration of sperm(80$\times$104, 40$\times$104, 20$\times$104, 10$\times$104, 5$\times$104, 3$\times$104, 1$\times$104/20${mu}ell$. The artificial insemination was succeeded but fertilization rate was very low(5~15%) compared to the natural mating and 59 normal youngs born from 60 females. Therefore, our findings suggest that it is possible to produce the great number of mice from the same orgin of male by artificial inseminatin. However, the lower pregnancy rate has to be solved to used broadly the artificial insemination in mouse.