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Capacitation and acrosome reaction differences of bovine, mouse and porcine spermatozoa in responsiveness to estrogenic compounds

  • Ryu, Do-Yeal;Kim, Ye-Ji;Lee, June-Sub;Rahman, Md. Saidur;Kwon, Woo-Sung;Yoon, Sung-Jae;Pang, Myung-Geol
    • Journal of Animal Science and Technology
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    • v.56 no.7
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    • pp.26.1-26.10
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    • 2014
  • Background: Endocrine disruptors are exogenous substance, interfere with the endocrine system, and disrupt hormonal functions. However, the effect of endocrine disruptors in different species has not yet been elucidated. Therefore, we investigated the possible effects of $17{\beta}$-estradiol (E2), progesterone (P4), genistein (GEN) and 4-tert-octylphenol (OP), on capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. In this in vitro trial, spermatozoa were incubated with $0.001-100{\mu}M$ of each chemical either 15 or 30 min and then assessed capacitation status using chlortetracycline staining. Results: E2 significantly increased capacitation and the acrosome reaction after 30 min, while the acrosome reaction after 15 min incubation in mouse spermatozoa. Simultaneously, capacitation and the acrosome reaction were induced after 15 and 30 min incubation in porcine spermatozoa, respectively. Capacitation was increased in porcine spermatozoa after 15 min incubation at the lowest concentration, while the acrosome reaction was increased in mouse spermatozoa after 30 min (P < 0.05). E2 significantly increased the acrosome reaction in porcine spermatozoa, but only at the highest concentration examined (P < 0.05). P4 significantly increased the acrosome reaction in bovine and mouse spermatozoa treated for 15 min (P < 0.05). The same treatment significantly increased capacitation in porcine spermatozoa (P < 0.05). P4 significantly increased capacitation in mouse spermatozoa treated for 30 min (P < 0.05). GEN significantly increased the acrosome reaction in porcine spermatozoa treated for 15 and 30 min and in mouse spermatozoa treated for 30 min (P < 0.05). OP significantly increased the acrosome reaction in mouse spermatozoa after 15 min (P < 0.05). Besides, when spermatozoa were incubated for 30 min, capacitation and the acrosome reaction were higher than 15 min incubation in E2 or GEN. Furthermore, the responsiveness of bovine, mouse and porcine spermatozoa to each chemical differed. Conclusions: In conclusion, all chemicals studied effectively increased capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. Also we found that both E2 and P4 were more potent than environmental estrogens in altering sperm function. Porcine and mouse spermatozoa were more responsive than bovine spermatozoa.

Effect of Garlic Powder of Mouse on the Stamina Improvement (마늘분말이 Mouse의 체력증강에 미치는 영향)

  • 박무현
    • Korean Journal of Plant Resources
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    • v.8 no.3
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    • pp.319-324
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    • 1995
  • The effect of garlic powder were investigated on health of stamina through mouse test. The swimming time of mouse was measured to determine the effect of garlic powder on stamina. When 2,000 and 200mg/kg of garlic were treated to mouse, swimming times were $124.8\pm61.1$ and $100.4\pm61min$, respectively. Those time were significantly longer than untreated control mouse which showed $67.1\pm15.5min$ of swimming time.

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A Study on the Short Break Time on VDT Work using EMG (근전도를 이용한 VDT 작업시 짧은 휴식시간에 관한 연구)

  • Kim, Yu-Chang;Lee, Jun-Pal
    • Journal of the Ergonomics Society of Korea
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    • v.26 no.4
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    • pp.41-47
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    • 2007
  • This paper suggested the best work conditions including short break time and the number of mouse clicks on the computer work for the prevention of MSDs on VDT work. Fatigue measures included EMG based parameters. The short break time conditions are grouped into 7, 15, and 30 seconds after every work for 10 min and the number of mouse clicks are divided into 10, 20, and 30 clicks/min. The result of the ANOVA of the shift value of %MVC(Maximum Voluntary Contraction) showed the following: 1) There was a considerable difference as regards to the break time except the number of mouse clicks on the upper trapezius muscle(p$<$0.05). The best conditions were shown in 15 sec after every 10 min and 30 clicks/min. 2) There were considerable differences as regards to the number of mouse clicks except the break time on the extensor digitorum muscle and extensor carpi ulnaris muscle(p$<$0.05). The best conditions were shown in 7 sec after every 10min and 10 clicks/min.

The Effects of Short Break Time and Mouse Clicks on the VDT Work by using Subjective Discomfort (VDT 작업시 짧은 휴식시간과 마우스 클릭이 주관적 불편도에 미치는 영향에 관한 연구)

  • Kim, Yu-Chang;Lee, Jun-Pal
    • Journal of the Korean Society of Safety
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    • v.23 no.2
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    • pp.30-36
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    • 2008
  • This paper suggests the best work conditions, including short break time and number of mouse clicks on a computer, to prevent MSDs(Musculoskeletal Disorders) on VDT(Visual Display Terminal) work. Discomfort measures are calculated according to the Borg's CR-10 Scale. The short break time conditions are grouped into 7, 15, and 30 seconds after every 10-minute work period and the number of mouse clicks are divided into 10 clicks/min, 20clicks/min, and 30clicks/min. The result of the ANOVA on the shift value of subjective discomfort shows the following: 1) Regarding the break time and the number of mouse clicks, there are statistical differences between the measured values for the neck and the wrist(p<0.05). 2) Regarding the number of mouse clicks, there are statistical differences between the measured values for the shoulder and the forearm(p<0.1). 3) Regarding the break time and the number of mouse clicks, there are no statistical differences between the measures values for the eyes, upper arms and back(p<0.1).

Effects of Somatic Cell Conditioned Medium on the Chymotrypsin Resistance of Mouse Oocytes (체세포배양액이 생쥐 난자의 Chymotrypsin에 대한 내성에 미치는 영향)

  • Kim, Sung-Rye;Chung, Hye-Won;Kim, Seong-Im;Kim, Hae-Kwon
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.207-216
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    • 1998
  • Certain types of somatic cells, as well as follicular cumulus cells associating with mammalian oocytes, are known to produce beneficial effects on in vitro fertilization and pre implantation development of mammalian eggs when they are present in oocyte culture medium. To investigate the nature of the effects of somatic cells, the resistance of mouse oocytes against chymotrypsin treatment was examined after culture within various cell conditioned media. When mouse oocytes matured for 17-18 hr in the presence of cumulus cells were treated with 1 % chymotrypsin, half of them remained still alive even after 240 min $(t_{50}>240.0)$. In contrast half of mouse oocytes cultured without cumulus cells underwent degeneration within 65.0 min $(t_{50}=65.0{\pm}13.2min)$ of the same treatment. To see if the effects were duc to the secretory products of cumulus cells, mouse cumulus cells were cultured for 20 hr in medium containing 0.4% BSA and the supernatant of culture medium (conditioned medium) was taken. After maturation in the cumulus cell conditioned medium, mouse oocytes exhibited $t_{50}=190.0{\pm}10.8$ min upon chymotrypsin treatment whereas half of oocytes cultured without conditioned medium degenerated within 25.5 min. Human granulosa cell conditioned medium gave similar effects such that oocytes matured in conditioned medium exhibited $t_{50}=183.3{\pm}19.1$ min while $t_50$ of control group oocytes was $60.0{\pm}6.8$ min, Oocytes matured in vero cell conditioned medium exhibited $t_{50}=196.7{\pm}8.8$ min. On the other hand, amniotic cell conditioned medium resulted in the chymotrypsin resistance of $t_{50}=80.0{\pm}8.4$ min which was not statistically different from the control value of $t_{50}=48.0{\pm}13.2$ min. Based upon these results, it is suggested that certain somatic cell types including cumulus cells might change the biochemical properties of mouse oocyte membrane during meiotic maturation as revealed by the enhanced resistance against chymotrypsin treatment. Such effects of somatic cells appear to be mediated via the secretory products rather than direct communication between somatic cells and oocytes.

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In Vitro Development of Mouse Parthenogenetic Embryos: Effect of Temperature before Oocyte Activation

  • Roh Sangho;Won Cheolhee;Min Byung-Moo
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.117-120
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    • 2005
  • This study was conducted to establish the optimal temperature condition before oocyte activation in B6m F1 mouse. In experiment 1, two embryo culture media (CZB vs KSOM) were evaluated for the development of activated mouse oocytes. Parthenogenetic embryos cultured in KSOM showed better blastocyst development than ones cultured in CZB $(56.2\%\;vs\;81.0\%\;p<0.01)$. Two-hour of pre-incubation before activation significantly reduced the number of hatched blastocysts in KSOM $(22.0\%\;versus\;8.8\%\;p<0.05)$. In experiment 2, recovered oocytes were pre-incubated at different temperature conditions before activation. The experimental groups were divided by 5 as follows. Group A: pre-incubation for 120 min at $37^{\circ}C$, Group B: pre-incubation at $37^{\circ}C$ for 90 min then at $25^{\circ}C$ for 30 min, Group C: pre-incubation at $37^{\circ}C$ for 60 min then at $25^{\circ}C$for 60 min, Group D: pre-incubation at $37^{\circ}C$ for 30 min then at $25^{\circ}C$ for 90 min, and Group E: pre-incubation at $25^{\circ}C$ for 120 min before activation. Group A $(67.6\%)$ and B $(66.7\%)$ showed better development to the blastocyst stage than other groups $(Group\;C:\;50.0\%\;Group \;D:\;49.2\%\;Group\;E:\;33.3\%,\;p<0.05)$. The present study indicates that the temperature before activation affects the development of B6D2 F1 mouse parthenogenetic oocytes and exposure to room temperature should be limited to 30-min when the oocytes are left in HEPES-buffered medium for micromanipulation.

Effects of Heating on Hydroxyl Radical-Generated Toxicity in Mouse Forebrain Tissue Culture

  • Lee, Jeong-Chae;Lim, Kye-Taek
    • Toxicological Research
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    • v.14 no.3
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    • pp.301-306
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    • 1998
  • This experiment was carrid out to know the effects of heating and serum on hydroxyl radicals in embryonic mouse forebrain (cerebrum) culture. The heating to mouse embryonic cerebrum cells in culture was done in a water bath at 43${\circ}C$ for 60min. After that, two supernatants were prepared at 20 hrs and 48 hrs respectively after heat treatment to the brain cells. To find out the heating effects on neuron cells, mouse cerebrum cells (13 embryonic day) were cultured in hydroxyl radical generation system composed of 20mU/ml glucose oxidase (GO system), using condition of normal culture media (MEM, 5% serum, 5% $CO_2$or supernatant prepared after heating at 43${\circ}C$ for 60 min in a water bath. Supernatant prepared at 20 hrs after heat treatment had a greater protective effects against hydroxyl radical than supernatant prepared at 48 hrs after heat treatment . Otherwise, the protective effect of serum against hydroxyl radicals in the cultured brain cells is higher than that in the heat treatment. These results indicated that serum in culture media reduced cytotoxicity of hydroxyl radicals in mouse forebrain culture, also that heat treatment showed the protective effects against hydroxyl radicals generated with 20mU/ml GO system in mouse forebrain culture.

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Effects of Aggregation Methods of Mouse Blastomeres on Aggregation Rate (생쥐 분리할구의 융합방법이 융합율 향상에 미치는 영향)

  • 최선호;정영채;김창근;정영호;윤종택;송학웅
    • Journal of Embryo Transfer
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    • v.9 no.1
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    • pp.111-116
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    • 1994
  • This study was carried out to investigate the aggregation rate of isolated mouse 2-, 4- and 8-cell stage blastomeres in phytohemagglutinin(PHA) solution. The morphologically normal embryos were collected from the oviduct of superovulated female mouse by flushing with M2 and the zona pellucida of embryos were removed with 0.5% pronase. The blastomeres were isolated by pipetting after plunging into Ca++-Mg++free PBS for 20 min. The result showed that aggregation rate in 0.5% (84.9~93.1%) was higher than that in 1.0% PHA(76.0~82.1%). Optimal aggregation time was 60min (83.9~100.0%) when compared with 30min (78.8~87.5%). Developmental to blastocyst in recombinated blastomeres was higher under conditions of 0.5% PHA solution and 60-min aggregation than that under other conditions.

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Time Courses of pCREB Expression after Dopaminergic Stimulation by Apomorphine in Mouse Brain

  • Jang, Choon-Gon;Lee, Seok-Yong;Lee, Han-Kyu;Suh, Hong-Won;Song, Dong-Keun
    • Archives of Pharmacal Research
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    • v.25 no.3
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    • pp.370-374
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    • 2002
  • Administration of dopamine agonist, apomorphine (2 mg/kg, s.c.), produces cage climbing behavior in mice that exhibit typical dopaminergic stimulation. The present study investigated the pCREB expression level in several brain regions following apomorphine treatment in order to determine whether the increased the dopaminergic activation produced by apomorphine accompanies the changes in pCREB immunoreactivity. A mouse brain was removed at 0min, 10 min, 30 min, 1 h, 2 h, 7 h, and 24 h after apomorphine treatment. The brain tissue was fixed by an intracardiac perfusion with ice-cold 4% paraformaldehyde in PBS. Immunohistochemical study was conducted using the ABC-DAB method. The data showed that the immunoreactivity of pCREB increased in the striatum, nucleus-accumbens, piriform cortex and the dentate gyrus of the hippocampus of a mouse brain 30 min after the apomorphine treatment. Increased immunoreactivity began to diminish 2 h after the apomorphine treatment in all the brain regions measured. The time course for the pCREB immunoreactivity was similar to the behavioral response induced by the apomorphine treatment. These results suggest that activation of the dopamine receptor is accompanied by an increase in pCREB expression in the mouse brain.

A Simultaneous NIRS-EEG Study of Seizure in the Mouse Brain

  • Lee, Seung-Duk;Lee, Min-Ah;Koh, Dalk-Won;Kim, Beop-Min;Choi, Jee-Hyun
    • Proceedings of the Optical Society of Korea Conference
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    • 2008.07a
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    • pp.159-160
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    • 2008
  • We measured hemodynamic responses of seizure in the mouse brain using frequencydomain near infrared spectroscopy (NIRS) and electroencephalogram (EEG). We adapted microfabricated optical holder for consistent contact of the optical fiber to the mouse brain. Our results show that the cerebral oxygenation and hemodynamics of mice can be stably monitored with EEG in the mouse brain.

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