• Korean Journal of Veterinary Research
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• v.32 no.2
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• pp.157-164
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• 1992

Changes in the both inward current and conductance of membrane by the fertilization were observed using the one microelectrode voltage clamp(or switch clamp) technique. Unfertilized eggs and both 1- and 2-cell stage eggs after fertilization were donated from the superovulated mouse (ICR, more than 6 weeks old) treated with PMSG(pregnant mare serum gonadotropin, Sigma) and HCG(human chorionic gonadotropin, Sigma) and naturally mated ones, respectively in this experiment. Membrane potential was held at -90mV and the voltage step was applied from -80mV to 50mV with interval of 10mV or 20mV for 300ms. since both of amplitudes and time courses in the membrane currents were various according to the states of cells and clamping condition, results were presented by their $averages{\pm}SEM$(standard mean error)and ratios or percentages. Inward currents began to appear in response to the step depolarization from -60mV and reached its maximum at -50mV. However, since the potential was not clamped evenly during the voltage step, current-voltage(I-V) relationship might be positively shifted 10 or 20mV. From the steady-state currents plotted in the I-V curve, outward rectification was markedly observed. Peak inward currents$(i_{in})$ at -50mV were $-0.62{\pm}0.23nA$(n=4),$-0.52{\pm}0.25nA$(n=5) and $-0.37{\pm}0.25nA$(n=6), in the 1-cell stage, 2-cell stage fertilized eggs and in the unfertilized eggs, respectively. Pure inward current (difference between steady-state and peak, $i_{in. pure}$) were $-1.01{\pm}0.23nA$, $-0.69{\pm}0.43nA$ and $-0.68{\pm}0.29nA$, respectively in the 1-cell stage fertilized eggs, unfertilized eggs and 2-cell stage fertilized eggs. These results suggested that the outward current in fertilized eggs of 2-cell stage was more increased than those in the unfertilized eggs. Pure inward currents in the all stages of eggs showed a similar fashion in the I-V relationship from -50mV to 50mV and reversal potential at 50mV. Time constant of inactivation$({\tau})$ in the inward current was decreased as the membrane potential was depolarized in the unfertilized and 2-cell stage eggs but in the 1-cell stage eggs t was not likely to be affected significantly. Slope conductances were 14.2nS, 8.9n5 and 7.7nS in the 1-cell, 2-cell stage fertilized eggs and the unfertilized eggs, respectively. Membranes between two cells within a zona pellucida seem to be electrical-connected in the 2-cell stage eggs from the observation made in the analysis for the electronic spread and decay to the current stimuli. Both of inward current and membrane conductance were increased after fertilization in the mouse eggs. Inward current seems to be carried by the same ion or through the same channels up to the 2-cell stage and ion that carried inward current was thought to play important function after fertilization in the mouse eggs.

• Toxicological Research
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• v.17
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• pp.197-199
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• 2001

Enhancement of antigen-specific IgE is a hallmark of allergic hyperresponsiveness, therefore it is necessary to adopt or develop a highly sensitive and specific assay for determination of allergen-specific IgE levels in vivo. In this presentation, we introduce an ELISA (enzyme linked immunosorbent assay) system developed to measure the levels of chicken egg ovalbumin (OVA)-specific IgE in serum. The ELISA method uses a commercially available purified rat anti-mouse IgE as a capture Ab and biotinylated OVA as a detection reagent. Avidin-peroxidase with its substrate is used for color development resulting in optical density measurement at 405 nm. The ELISA system produces a highly sensitive dose-response relation-ship between optical density levels and the dilution titer of the OVA-IgE standard serum but no cross-reaction with unrelated IgE or IgG. It is believed that the system is an Efficient tool to delineate an adjuvant effect of environmental pollutants on development of asthmatic and atopic responses.

• Journal of Animal Science and Technology
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• v.57 no.4
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• pp.16.1-16.7
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• 2015

A well-characterized sperm specific protein of the Member of immunoglobulin superfamily, IZUMO1, has crucial role in fertilization by mediating sperm binding to the egg plasma membrane in the mouse. However little is known about IZUMO1 in bovine. Here, we describe the molecular cloning and expression analysis of bovine IZUMO1 (bIZUMO1). RT-PCR and Western blot analysis of the bovine tissues indicated that bIZUMO1 was specifically expressed in the testis and sperm, Furthermore, the result of our biotinylation assay from ejaculated bovine sperm strongly suggest the assumption that bIZUMO1 is localized on the cell surface. These data imply the potential role of bovine IZUMO1 in mammalian fertilization.

• Development and Reproduction
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• v.23 no.1
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• pp.1-9
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• 2019

The ability of oocytes to undergo normal fertilization and embryo development is acquired during oocyte maturation which is transition from the germinal vesicle stage (GV), germinal vesicle breakdown (GVBD) to metaphase of meiosis II (MII). Part of this process includes redistribution of inositol 1, 4, 5-triphosphate receptor (IP3R), a predominant $Ca^{2+}$ channel on the endoplasmic reticulum membrane. Type 1 IP3R (IP3R1) is expressed in mouse oocytes dominantly. At GV stage, IP3R1 are arranged as a network throughout the cytoplasm with minute accumulation around the nucleus. At MII stage, IP3R1 diffuses to the entire cytoplasm in a more reticular manner, and obvious clusters of IP3R1 are observed at the cortex of the egg. This structural reorganization provides acquisition of $[Ca^{2+}]_i$ oscillatory activity during fertilization. In this review, general properties of IP3R1 in somatic cells and mammalian oocyte are introduced.

• The Korean Journal of Zoology
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• v.13 no.3
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• pp.68-74
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• 1970

In the previous studies, the present author found that high proportion of the follicular oocytes from mouse and rabbit ovaries are able to resume their maturation division in the anterior chamber of the eye in which they have been incubated by auto- or homoplastic transplantation. Especially in the case of the homoplastic transplantation, it was known that no trouble has been detected in the process of resumption of the oval maturation in particular connection with the antigen-antibody reaction between donor and recipient. These findings provide a possibility that the follicular oocytes from various animals would be matured in the eye even after the xenoplastic transplantation. Under such an assumption, the present studies were performed to examine the behavior of the follicular oocytes in the eye chamber of the animals of different species. For the donor of the follicular oocytes, domestic rabbits, albino rats of Sprague-Dowley strain, and albino mice of A-strain bred in our laboratory were used. The oocytes obtained from the ovarian follicules were introduced to the anterior chamber of the eye of different species of animals, with an exception of rabbit in which only the female animals were used as a recipient. The procedures of collection of ova, introduction to the eye, harvest from the eye ball, fixation, and staining were the same as mentioned in the previous reports (Cho, 1967b; Cho and Kim, 1968). The conclusions obtained are summarized as below. 1. The rabbit follicular oocytes are able to mature in the eye chambers of both male mouse and rat, although the proportion of the maturation is lower than when they are incubated autoplastically in the eye. When the ova were incubated in the male mouse eye for 24 hours, 21 per cent of them showed chromosomes at metaphase I and II, whereas the rate was 32 per cent when they were incubated in the eye of the male rat. These are apparently low comparing to the rate of 52 per cent of autoplastic transplantation. 2. When rat follicular oocytes were transferred into the mouse eye chamber and recovered after 24 hours, 43 per cent of them produced the mataphase I and II chromosomes. This proportion was higher than the result of the homoplastic transplantation which yielded 23 per cent of the ova on maturation. 3. The most striking result was found in the experiment with mouse follicular oocytes. Seventy-six per cent of the oocytes resumed their maturation division within 24 hours after they were transferred into the male rat eye chamber, and this figure was significantly high compared to the result o 55 per cent obtained by the homoplastic transplantation. In the rat eye, the induction of the degenerative ova also was low (19%). On the other hand, the proportion of the oval maturation decreased to 45 per cent, while that of degeneration increased 33 per cent when they were incubated in the eye of the female rabbit. 4. It was apparent from the present experiments that the follicular oocytes can reveal their activation to maturation in the eye chamber which contains aqueous humor which is known to be composed of low protein content and of very little gamma-globulin which acts as an antibody(Oser, 1965), and that it shows higher osmolarity than blood serum(Levene, 1958). Taking these properities into consideration the humor may provide unfavourable environment to the cells and tissues incubated in. However, it could be noteworthy finding that only the follicular oocytes in the eye of the different species can grow in healthy condition although the maturation rates are varied with the animal species. The fact that the rabbit follicular oocytes show the lower proportion in maturation may be due to the greater amount of the yolk granules in the egg cytoplasm than those in the mouse and rat oocytes. That the mouse oocytes incubated in the eye of the rat mouse and rat oocytes. That the mouse oocytes incubated in the eye of the rat resumed their maturation process in greater proportion would e explained by the fact that the rat eye chamber particularly provides the better environment to the mouse oocytes than the eye chamber of mouse does.

• Korean Journal of Food Science and Technology
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• v.38 no.1
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• pp.93-98
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• 2006

Effects of anti-Helicobacter pylori IgY powder on H. pylori infection were evaluated 3 and 7 weeks after powder feeding by urease, PCR, and histological tests, and specific IgG assay of murine gastric tissue using mouse model. To produce anti-H. pylori IgY powder, laying hens were immunized with H. pylori prior to egg yolk harvest. C57BL/6 mice showing high response to H. pylori were infected with H. pylori and fed with the anti-H. pylori IgY powder. In urease and PCR tests, urease activity and gene count of anti-H. pylori IgY powder-fed group significantly decreased in comparison with control. Histological results indicated anti-H. pylori IgY powder effectively protected mice from H. pylori.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.377-383
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• 1997

The effects of embryo number and incubation volume on the development of mouse embryos were evaluated. The growth rate of two-cell mouse embryos to attached blastocyst stage and the growth rate of blastocysts to early somite stage were assessed after culture in different incubation volumes and embryo densities. Embryos were collected from ICR female mice superovulated with pregnant mare serum gonadotropin and human chorionic gonadotropin and mated by ICR males. In experiment 1, groups of one, five, ten, twenty 2-cell embryos were cultured in 10-, 50-, 500-, 1000-${\mu}l$ drops of BWW media under mineral oil at $37^{\circ}C$ in a humidified atmosphere of 5% $CO_{2}$ and 95% air. As the incubation volume decreased, significantly (p<0.05) higher rates of embryos reached morular and blastocyst stage on day 3 and 4 culture, respectively. In experiment 2, groups of one, five, ten, twenty blastocysts were cultured in 1- and 2-ml volumes of CMRL 1066 media under same condition as in experiment 1. However the reverse was the result. Decreasing the number of embryos incubated per volume from 1 to 20 significantly (p<0.05) increased the number of blastocysts reaching the late egg cylinder (LEC) and early somite (ES) stage on day 6 and 8 culture, respectively, regardless of incubation volume. Blastocysts cultured in 2ml had higher (p<0.05) development rates to LEC and ES stage on day 6 and 8 culture, respectively, than embryos cultured in 1ml. Our results suggest that the effects of embryo number and incubation volume on the development of mouse embryos are stage specific and the shifting point was between hatching and EEC stage.

• Parasites, Hosts and Diseases
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• v.55 no.1
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• pp.21-29
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• 2017

Schistosoma haematobium is a biocarcinogen of human urinary bladder (UB). The present study investigated developing UB cancer mouse model by injecting S. haematobium eggs into the bladder wall and introduction of chemical carcinogens. Histopathological findings showed mild hyperplasia to epithelial vacuolar change, and high grade dysplasia. Squamous metaplasia was observed in the S. haematobium eggs+NDMA group at week 12 but not in other groups. Immunohistochemistry revealed significantly high expression of Ki-67 in urothelial epithelial cells of the S. haematobium eggs+BBN group at week 20. The qRT-PCR showed high expression of p53 gene in S. haematobium eggs group at week 4 and S. haematobium eggs+BBN group at week 20. E-cadherin and vimentin showed contrasting expression in S. haematobium eggs+BBN group. Such inverse expression of E-cadherin and vimentin may indicate epithelial mesenchymal transition in the UB tissue. In conclusion, S. haematobium eggs and nitrosamines may transform UB cells into squamous metaplasia and dysplasia in correlation with increased expression of Ki-67. Marked decrease in E-cadherin and increase in p53 and vimentin expressions may support the transformation. The present study introduces a promising modified animal model for UB cancer study using S. haematobium eggs.

• Archives of Pharmacal Research
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• v.17 no.4
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• pp.209-212
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• 1994

Mophological normal of unfertilized oocytes, which was collected 12-14 hours after human Chorionic Gonadotropin(jCG) injection, was not influenced by chronically adiministration of cocaine for 2 weeks in mice. Proportion of normal unfertilized oocytes in non-cocaine treated group (control), `0 mg/kg and 20 mg/kg cocaine treated group based on body weight with subcutaneous(s.c.) daily injection of cocaine for 2 weeks were 92.9%, 85.6% and 90.9%, respectively. There is no significant difference between control and cocaine treated groups. Two to 8 cell stage embryos collected 24-48 hours post hCG in control group were 66.7%, whereas, 10 mg/kg and 20 mg/kg groups treated with cocaine was 12.5% and 27.3% respectively. Although control and treated groups are significantly different (p<0.05) the developmental score of 2 to 8 cell stage embryos collected at 24-48 hours post HCG, there is no difference between 10 mg/kg and 20 mg/kg treated with cocaine groups. These results indicated that the normal embryos of the roups of cocaine administration were significantly amested when compared with that of control group. The proportion of 2 to 8 cell stage embryo reaching the blastocyst stage, which were cultured 48-52 hours with 5% $Co_2$ in air at $37^{\circ}C$, were 93.9% in control group and, 70.4% and 71.9% in each 10 mg/kg and to blastocyst in vitro culture was significantly limited embryos obtained from cocanized mice compared with those of control mice. These results suggest that episode of cocaine intoxication can cause impaiment of early embrygenesis in the mouse.

• Journal of the korean academy of Pediatric Dentistry
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• v.28 no.4
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• pp.709-727
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• 2001

The pattern of programmed cell death(PCD) has been examined during the early developmental period of development in mouse embryos, from embryonic day 4.5(E4.5) to E11.5 Embryos from Balb/c breedings were harvested at various embryonic stages between E4.5 and El1.5. Cell death was analysed by in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL) staining in tissue sections and whole embryos. At the blastocyst stage(E4.5), a very few apoptotic cells were found in the inner cell mass of the blastocyst. In the early egg cylinder stage(35.0-5.5), a few apoptotic cells were detected in the embryonic ectoderm, the embryonic endoerm and the proamniotic cavity. In the advanced egg cylinder stage(E5.5-6.5), TUNEL-posifive cells were observed in the extra-embryonic ectoderm and extra-embryonic endoderm as well as in the embryonic ectoderm, embryonic visceral endoderm and proamniotic cavity. In the streak stage(E6.75-7.75), many TUNEL-positive cells were found in the ectoplacental cone. In contrast, only very few apoptotic cells were found in the chorion and extra-embryonic endoderm in extra-embryonic regions. In intra-embryonic region, a few apoptotic cells were randomly found in the embryonic ectoderm, mesoderm and visceral endoderm. At the early somitogenesis stage(E8.0-8.5), most apoptotic cells were observed in the most cranial portion of neural fold (neural ectoderm and adjacent ectoderm). At the mid somitogenesis stage(39.0-9.5), the otic placode first showed TUNEL-positive at this stage. Small number of TUNEL-positive cells were also first seen around optic placode and branchial arches. Three streams of TUNEL-positive cells were clearly seen in the cranial region at 59.5-9.75. At E10.5, apoptotic cells were localized in the developing eye, the junctional portion of medial nasal, lateral nasal and maxillary processes, the lateral portion of branchial arches, the junction of bilateral mandibular processes, and apical ectodermal ridges of limb buds. At E11.5, apoptotic cells were noticeably decreased in most area, except the developing limbs and several somites in the tail region. In this study, the global temporospatial pattern of PCD throughout early development of mouse embryos was discussed. It may provide the basis for further studies on its role in the morphogenesis of the embryo.