• 한국발생생물학회지:발생과생식
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• 제17권4호
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• pp.461-467
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• 2013

Nesfatin-1/NUCB2, which is associated with the control of appetite and energy metabolism, was reported for the first time to be expressed in the hypothalamus. However, recent studies have shown that nesfatin-1/NUCB2 was expressed not only in the hypothalamus, but also in various tissues including digestive and reproductive organs. We also demonstrated that nesfatin-1/NUCB2 was expressed in the reproductive organs, pituitary gland, heart, lung, and gastrointestinal tract of the adult mouse. However, little is known about nesfatin-1/NUCB2 expression in fetal and neonatal mice. Therefore, we examined here the distribution of nesfatin-1/NUCB2 in various organs of fetal and neonatal mice and compared them with the distribution in adult mice. As a result of immunohistochemical staining, nesfatin-1/NUCB2 protein was expressed relatively higher in the lung, kidney, heart, and liver compared to other organs in the fetus. Western blot results also showed that nesfatin-1/NUCB2 protein was detected in the lung, kidney, heart, and stomach. Next, we compared the expression levels of nesfatin-1/NUCB2 mRNA in the fetus and neonate with the expression levels in both male and female adult mice. The expression levels in heart, lung, stomach, and kidney were higher compared with other organs in fetal and neonatal mice and in both male and female adult mice. Interestingly, the expression of nesfatin-1/NUCB2 mRNA in the kidney was dramatically increased in male and female adult mice compared to fetal and neonatal mice. These results indicate that nesfatin-1/NUCB2 may regulate the development and physiological function of mouse organs. In the future, we need more study on the function of nesfatin-1/NUCB2, which is highly expressed in the heart, lung, and kidney during mouse development.

• Clinical and Experimental Reproductive Medicine
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• 제35권1호
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• pp.39-48
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• 2008

목 적: 본 연구에서는 신생 생쥐 고환으로부터 다분화능 생식줄기세포주 (MGSCs)를 확립하고, 배아체 형성을 통한 삼배엽성 세포로의 분화 가능성을 확인하고자 하였다. 연구방법: 고환에서 유래한 MGSCs를 확립하기 위하여 생후 $2{\sim}3$일된 생쥐 고환 조직으로부터 세포들을 분리하여 1% FBS를 첨가한 생쥐 배아줄기세포주 배양조건에서 배양하였다. MGSCs 콜로니가 형성된 후에는 배양액의 FBS의 농도를 15%로 높였다. 이러한 과정으로 확립된 MGSCs의 미분화 및 분화 특성을 배아줄기세포주와 비교, 분석하였다. 결 과: 신생 생쥐 고환 조직에서 수획한 세포들로 실시한 9번의 배양실험에서 2개의 MGSCs 세포주를 확립하였다. MGSCs 세포주와 생쥐 배아줄기세포 모두에서 미분화 표지인자인 Thy-1, Oct-4, Nanog, Sox2의 발현과 alkaline phosphatase 활성을 관찰할 수 있었으며, MGSCs의 미세구조 또한 생쥐 배아줄기세포와 유사하였다. MGSCs에서 형성된 배아체에서 삼배엽성 표지유전자의 발현을 확인하였다. 결 론: 본 연구의 결과는 배아줄기세포의 윤리적인 문제점을 극복할 수 있는 고환 유래의 다분화능 MGSCs가 생물공학과 재생의학에서 효율적으로 이용될 수 있는 가능성을 보여준 것으로 생각된다.

• 한국환경보건학회지
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• 제48권2호
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• pp.106-115
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• 2022

Background: Current research suggests that humans are exposed to microplastics through consumption of foods and beverages, the airway route, and a variety of other means. Objectives: We evaluated oxidative stress and inflammation from polyethylene microplastics (PE-MPs) in the neonatal liver through intragastric administration or intratracheal instillation in pregnant mice. Methods: PE-MPs were administered from gestational day 9 to postnatal day 7. The intragastric administration group (0.01 mg/mouse/day or 0.1 mg/mouse/day) and intratracheal instillation group (6 ㎍/mouse/day or 60 ㎍/mouse/day) of PE-MPs were administered. After sacrifice, the oxidative stress and inflammation of the neonatal livers were measured. Results: As a result of the oxidative stress caused by PE-MPs in the neonatal livers, glutathione peroxidase decreased in a concentration-dependent manner in the intragastric administration group compared to the control group and intratracheal instillation decreased in high concentration PE-MPs. The catalase level increased at high concentrations of intragastric administration and intratracheal instillation. To confirm the level of inflammation caused by PE-MPs, monocyte chemoattractant protein-1 and tumor necrosis factoralpha were increased compared to the control group except for intratracheal intilation-high concentration PEMPs. The C-reactive protein level was decreased by intragastric administration compared to the control group and intratracheal instillation was increased compared to the control group. Conclusions: Despite the difficulty in comparing the toxic intensity between intragastric administration and intratracheal instillation of PE-MPs, our study revealed that oxidative stress and inflammation were induced in the neonatal liver. However, it is necessary to evaluate the toxic effects of microplastics on various organs as well. Overall, the present study indicates that the evaluation of toxic effects of long-term microplastic exposure, potential of microplastic toxicity on next-generation offspring and toxicity mechanism in human should be considered for further investigations.

• Clinical and Experimental Pediatrics
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• 제62권12호
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• pp.444-449
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• 2019

Background: Sixty percent of infants with severe neonatal hypoxic-ischemic encephalopathy die, while most survivors have permanent disabilities. Treatment for neonatal hypoxic-ischemic encephalopathy is limited to therapeutic hypothermia, but it does not offer complete protection. Here, we investigated whether hypoxia-inducible factor (HIF) promotes cell survival and suggested neuroprotective strategies. Purpose: HIF-1α deficient mice have increased brain injury after neonatal hypoxia-ischemia (HI), and the role of HIF-2α in HI is not well characterized. Copper-zinc superoxide dismutase (SOD)1 overexpression is not beneficial in neonatal HI. The expression of HIF-1α and HIF-2α was measured in SOD1 overexpressing mice and compared to wild-type littermates to see if alteration in expression explains this lack of benefit. Methods: On postnatal day 9, C57Bl/6 mice were subjected to HI, and protein expression was measured by western blotting in the ipsilateral cortex of wild-type and SOD1 overexpressing mice to quantify HIF-1α and HIF-2α. Spectrin expression was also measured to characterize the mechanism of cell death. Results: HIF-1α protein expression did not significantly change after HI injury in the SOD1 overexpressing or wild-type mouse cortex. However, HIF-2α protein expression increased 30 minutes after HI injury in the wild-type and SOD1 overexpressing mouse cortex and decreased to baseline value at 24 hours after HI injury. Spectrin 145/150 expression did not significantly change after HI injury in the SOD1 overexpressing or wild-type mouse cortex. However, spectrin 120 expression increased in both wild-type and SOD1 overexpressing mouse at 4 hours after HI, which decreased by 24 hours, indicating a greater role of apoptotic cell death. Conclusion: HIF-1α and HIF-2α may promote cell survival in neonatal HI in a cell-specific and regional fashion. Our findings suggest that early HIF-2α upregulation precedes apoptotic cell death and limits necrotic cell death. However, the influence of SOD was not clarified; it remains an intriguing factor in neonatal HI.

• Clinical and Experimental Reproductive Medicine
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• 제26권2호
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• pp.219-223
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• 1999

Ovarian development of the vitrified neonatal ovaries after orthotopical transplantation into the ovariectomized adult recipient mouse were observed. Ovaries were collected from the neonatal females on day of birth and grouped for fresh, vitrification for 1-minute, and 3-minute. Vitrified and thawed neonatal ovaries were orthotopically transplanted into ovarian bursa of the adult mice from which endogenous ovaries have removed just prior to the transplantation (1 minute: n=25; 3 minutes n=23). Fresh ovarian tissue transplanted (n=25) mice were included as control groups. Returning of the estrus cycles and the survival and development of the transplanted ovaries were evaluated. Intact ovaries from neonatal, and four weeks old mice were used for comparison of the ovarian development as in vivo-developed control. From 2 weeks after transplantation, 64%, 36%, and 75% of the transplanted mice showed return of the estrus cycles in fresh, 1-minute, and 3-minute groups, respectively. Four weeks after transplantation, all mice were sacrificed and ovarian tissues were recovered for histological analysis. 57.1%, 33.3%, and 64.7% mice in fresh, 1-minute, and 3-minute groups, respectively, had survived ovaries with follicles at various stages of growth from primordial to preovulatory follicles. Corpus lutea were also observed. Results of the present study suggest that 1) normal folliculogenesis has initiated in vivo after vitrification, and 2) the vitrification may be used as a preservation method for ovarian tissues for establishment of ovarian tissue bank.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2003년도 제3회 발생공학 국제심포지움 및 학술대회
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• pp.63-63
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• 2003

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2003년도 제45차 추계학술대회
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• pp.144-145
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• 2003

• BMB Reports
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• 제39권4호
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• pp.426-431
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• 2006

Embryonic stem cells (ESCs), representing a population of undifferentiated pluripotent cells with both self-renewal and multilineage differentiation characteristics, are capable of spontaneous differentiation into cardiomyocytes. The present study sought to define the kinetic characterization of lactate dehydrogenase (LDH) and creatine kinase (CK) of ESC- and neonatal-derived cardiomyocytes. Spontaneously differentiated cardiomyocytes from embryoid bodies (EBs) derived from mouse ESC line (Royan B1) and neonatal cardiomyocytes were dispersed in a buffer solution. Enzymes were extracted by sonication and centrifugation for kinetic evaluation of LDH and CK with spectrophotometric methods. While a comparison between the kinetic properties of the LDH and CK of both groups revealed not only different Michaelis constants and optimum temperatures for LDH but also different Michaelis constants and optimum pH for CK, the pH profile of LDH and optimum temperature of CK were similar. In defining some kinetic properties of cardiac metabolic enzymes of ESC-derived cardiomyocytes, our results are expected to further facilitate the use of ESCs as an experimental model.

• 대한한의학회지
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• 제21권2호
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• pp.79-86
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• 2000

Objectives and Methods : In order to elucidate toxic mechanism of myocardial damage and protective effect of myrrha water extract against cytotoxic effect of xanthine oxidase/hypoxanthine(XO/HX), cardioprotective effect of myrrha water extract was examined by MTT assay, LDH (Lactate Dehydrogenase) activity and heart beating rate after cultured myocardial cells derived from neonatal mouse were treated with various concentration of XO/HX, a free radical. Results : XO/HX induced a decrease of cell viability, an increase in the amount of LDH, and a decrease of heart beating rate on cultured myocardial cells in a dose-dependent manner. In cardioprotective effect of myrrha water extract, it showed a decrease in the amount of LDH and an increase of heart beating rate on cultured myocardial cells damaged by XO/HX. Conclusions : From the above results, it is suggested that XO/HX showed toxic effect in cultured myocardial cells derived from neonatal mouse and that myrrha water extract is very effective in the prevention of XO/HX-induced cardiotoxicity.

• The Korean Journal of Pain
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• 제14권2호
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• pp.136-141
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• 2001

Background: Sodium salicylate (SS) is a nonsteroidal anti-inflammatory drug (NSAID) for the treatment of neuralgia or pain from rheumatoid arthritis. When abused or used in excess, SS can induce cytotoxicity. The present study examined whether SS has a neurotoxic effect. Methods: Cell viability was examined by MTT [3-(4,5-dimethylthiazol-2,5-dipheny ltetrazolium bromide] assay and Sulforhodamine (SRB) assay after cultivating dorsal root ganglion (DRG) neurons derived from neonatal mouse. These cells were treated with various concentrations of SS for 24 hours. In addition, the amount of protein synthesis against SS was measured in these cultures. Results: Cell viability (20, $40{\mu}g/ml$ SS) significantly decreased in a dose-dependent manner. Additionally, SS inhibited protein synthesis after the exposure of cultured mouse DRG neurons to $30{\mu}g/ml$ of SS for 24 hours. Conclusions: The present study suggests that SS is toxic in cultured DRG neurons derived from neonatal mouse by decreasing cell viability and the amount of protein synthesis.