• Title/Summary/Keyword: induced maturation

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The Functional Role of Maturation Promoting Factor in the Two-cell Embryos (생쥐 2-세포기 배아에서 성숙유도물질의 기능적 역할)

  • 강해묵;이대기
    • The Korean Journal of Zoology
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    • v.36 no.2
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    • pp.277-284
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    • 1993
  • A possible involvement of maturation promoting factor (nfPF) in the two-cell block phenomenon was studied by fusion experiments. Germinal vesicle (GlF) ooeyte was fused with a blastomore from late or blocked 2-cell mouse embryos. and germinal vesicle breakdoum (GVBD) of fused GV oocvtes in the presence of dbcAMP (100$\mu$g/ml) was scored as an index of MPF aniviD. GnD was induced approximately 30% by fusion of a blastomere derived from late 2-cell embryos, but not from blocked 2-cell embryos. The rate of GVBD was changed when GV oocyte was fused with a blastomere from late 2-cell embryos which were treated with u-amanitin, puromvcin or colcemid before and after hsion: Treatment of late 2-cell embryos with puromycin (50 Is/mll but not with u-amanitin (100 Is/ml) clearly inhibited GVBD, indicating that do novo protein synthesis maw be required for the appearance of MPF activity in late 2-cell embryos. Treatment of late 2-cell embryos w기h colcemid (0.1 Is/mll doubled GVBD, presumably due to the maintenance of metaphase or mitotic phase. SDS-PAGE and twoiimensional electrophoresis revealed that there was no difference in protein synthetic pattern in late and blocked 2-cell embryos, but three phosphoproteins with 27, 35 and 46 M)a, presumsblv M-phase components were phosphorylated in late 2-cell embryos but not in blocked 2-cell embryos. It seems then that MPF activity is closely related to phosphorylstion of M-phase components in late 2-cell embryos.

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Protocadherin-7 contributes to maintenance of bone homeostasis through regulation of osteoclast multinucleation

  • Kim, Hyunsoo;Takegahara, Noriko;Walsh, Matthew C.;Ueda, Jun;Fujihara, Yoshitaka;Ikawa, Masahito;Choi, Yongwon
    • BMB Reports
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    • v.53 no.9
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    • pp.472-477
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    • 2020
  • Osteoclasts are hematopoietic-derived cells that resorb bone. They are required to maintain proper bone homeostasis and skeletal strength. Although osteoclast differentiation depends on receptor activator of NF-κB ligand (RANKL) stimulation, additional molecules further contribute to osteoclast maturation. Here, we demonstrate that protocadherin-7 (Pcdh7) regulates formation of multinucleated osteoclasts and contributes to maintenance of bone homeostasis. We found that Pcdh7 expression is induced by RANKL stimulation, and that RNAi-mediated knockdown of Pcdh7 resulted in impaired formation of osteoclasts. We generated Pcdh7-deficient mice and found increased bone mass due to decreased bone resorption but without any defect in bone formation. Using an in vitro culture system, it was revealed that formation of multinucleated osteoclasts is impaired in Pcdh7-deficient cultures, while no apparent defects were observed in differentiation and function of Pcdh7-deficient osteoblasts. Taken together, these results reveal an osteoclast cell-intrinsic role for Pcdh7 in maintaining bone homeostasis.

Induced Ovulation and Histological Changes of the Oocytes according to HCG and Trout Pituitary Extract in the Korean Loach, Misgurnus anguilicaudatus (한국산 미꾸라지에 있어서 HCG나 송어의 뇌하수체에 의한 배란유기와 난모세포의 조직학적 변화)

  • 윤종만;이상목;박홍양
    • Korean Journal of Animal Reproduction
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    • v.11 no.3
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    • pp.170-180
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    • 1987
  • This study was undertaken to determine the effects of HCG and trout pitutary on sexual maturation and ovulation in the Korean loach, Misgurnus anguillicaudatus. Sexual maturation was observed in the fish treated with 250+125 IU HCG within 2-4 days following injections. Total ovary weight and GSI were significantly greater in the hormone-treated fish than in the control fish. Furthermore, in the hormone-treated fish body weight increased by about 4-9 percent in 2 days. The water content of the ovulated eggs of the fish treated with 250+125 IU HCG was also greater than that in the control. Changes in both macroscopic & microscopic appearnce of Korean loach ovaries are describled, as well as changes in the gonadosomatic index. Oocytes pass through seven cytologically changes in the relative stages. By examining these stages, it was found that oogenesis occures in two broad phases. The previtellogenic phasebegins as a new oogonia continues to arise following spawning. These rapidly develop into early perinucleolus oocytes, which in turn develop into resting stage oocytes. Most oocytes remain in this stage when they develop into late perinucleolus oocytes. The vitellogenic phase begins as these late perinucleolus oocytes become transformed into early maturing oocytes through the accumulation of yolk. The cytoplasm completely fills with yolk as oocytes reach the late maturing stage. Shortly before spawning the final hyaline stage of developing is reached. Changes in the microscopic appearances of the ovaries were well correlated with changes in both gonadosomatic index and macroscopic appearance.

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Effect of Phytoecdysteroid on Pure Breed Performance of Silkworm Bombyx mori L.

  • Trivedy, Kanika;Dhar, Anindita;Kumar, S.Nirmal;Nair, K.Sashindran;Ramesh, M.;Gopal, Nisha
    • International Journal of Industrial Entomology and Biomaterials
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    • v.7 no.1
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    • pp.29-36
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    • 2003
  • Phytoecdysteroids with moulting hormone (MH) activity induce different responses in silkworms when used on different day of final instar, which can be manipulated for maximum benefit like early and uniform spinning behaviour, reducing crop loss and to increase cocoon yield. The results showed that application of this hormone on seed crop viz., CSR2, CSR4 and BL44 and BL67 in early stage of 5$^{th}$ instar i.e., at 72 hrs and 96 hrs though induced early and uniform spinning behaviour, there was an adverse effect by 9-36% on the economic characters like cocoon yield, cocoon weight, cocoon shell weight and also on fecundity etc. Application of this hormone in late stage of $5^th$ instar i.e., at the onset of spinning showed non-significant variations in some of the characters like cocoon weight, cocoon shell weight, cocoon shell ratio and fecundity. The physiological implications of phytoecdysteroid in hastening the maturation events and synchronization of spinning activities in different breeds are discussed.

Immunomodulatory Effects of Eckol, a Pure Compound of Ecklonia Cava, on Dendritic Cells

  • Kim, Mi-Hyoung;Joo, Hong-Gu
    • IMMUNE NETWORK
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    • v.6 no.4
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    • pp.199-203
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    • 2006
  • Background: Eckol purified from Ecklonia cava, a brown alga has been known to have cytoprotective effects on some cell lines against oxidants and ionizing radiation. However, there is no study about the effects of eckol on immune cells. Methods: Bone marrow (BM)-derived dendritic cells (DCs) were used to demonstrate the immunomodulatory effects of eckol on DCs, such as viability, the expression of surface markers, allogeneic stimulating capacity using MTI, flow cytometric, $^3H$-thymidine incorporation assay. Results: Eckol did protect DCs against cytokine withdrawal-induced apoptosis in a concentration dependent manner based on MTT assay. And also, it increased the expression of MHC class II and CD86 (B7.2) molecules, maturation markers, on the surface of viable DCs gated in FACS analysis. Furthermore, eckol-treated DCs stimulated the proliferation of allogeneic $CD4^+$ T lymphocytes compared to imDCs in $^3H$-thymidine incorporation assay. $CD4^+$ T lymphocytes activated with eckol-treated DCs produced the larger amount of IFN-${\gamma}$ and IL-4 than those cells with imDCs. Conclusion: Taken together, we demonstrate in this study that eckol, a pure compound of Ecklonia cava, may modulate the immune responses through the phenotypic and functional changes of DCs.

Activation of MAP Kinase during Maturation in Porcine Ooctyes (돼지 미성숙란의 체외배양시 MAP Kinase의 활성)

  • 장규태;박미령;윤창현
    • Korean Journal of Animal Reproduction
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    • v.22 no.3
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    • pp.265-276
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    • 1998
  • In an attempt to evaluate the function of MAP kinase of porcine oocytes and to develop a method of assessment for kinase activity, we used MBP as a substrate to detect the MAP kinase activity of porcine oocytes matured in in vitro. The MAP kinase which had lower activity during the first 20 hours of culture started to show an increased amount of activity at 25 hours at which a collapse in nuclear membrane was induced. Significant (P<0.05) a, pp.ared at 30 hours of being cultured. The gel phosphorylation method, MBP which has been known to be a substrate for kinase such as cdc2 kinase, was phosphorylated at two positions corresponding to ERK 1 (44kDa) and ERK2 (42 kDa) which are known as mammalian MAP kinase. The existence of MARKK and MAP kinase were identified with western blotting at 0 hour culture of immature GV oocytes. The amount of those proteins did not increase during 40 hours of culture, which suggest that the increase of MAP kinase activity was caused by phosphorylaton rather than due to change in protein amount. MAPKK and MAP kinase were shown to be dephosporylated with deactivated at M 1 stage by inhibition of protein synthesis with cycloheximide added at the strat following the cultrue. We have reulsts that indicate the existedence of MAP kinase cascade which was activated simultaneously with start of porcine oocyte maturation (GVBD).

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In Vitro Growth of Preantral Follicle and Maturation of Intrafollicular Oocyte from Aged Mice

  • Yoon, Jung-Ah;Choi, Jung-Kyu
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.1
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    • pp.35-39
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    • 2019
  • This study aimed to recover the ovarian function through in vitro culture of preantral follicles from aged mice. First, we isolated the preantral follicles from ovaries of sixty-seven-week old B6D2F1 mice with decreased fecundity to know how many follicles were present in them, which was 6 preantral follicles including 2 primary, 2 early secondary and late secondary follicles from 8 aged mice. It was confirmed that a few follicles (~2) were present in aged mice through histological analysis compared to adult mice as control. The 9 days of in vitro culture of preantal follicles showed in vitro growth and induced maturation after treatment with hCG (2.5 IU/mL) and EGF (5 ng/mL). Cumulus cells in the cumulus-oocyte complexes (COCs) were removed using hyaluronidase and oocytes at the germinal vesicle (GV) and GV breakdown (GVBD) were obtained from preantral follicle culture of aged mice in vitro. In conclusion, these observations demonstrated that there still were a few preantral follicles in the ovaries of 67 week-old mice, which we were able to culture in vitro and oocytes were obtained from them. This study proposed an in vitro culture system using preantral follicle as a therapeutic strategy for fertility preservation in humans for assisted reproductive medicine.

Insulin enhances neurite extension and myelination of diabetic neuropathy neurons

  • Pham, Vuong M.;Thakor, Nitish
    • The Korean Journal of Pain
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    • v.35 no.2
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    • pp.160-172
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    • 2022
  • Background: The authors established an in vitro model of diabetic neuropathy based on the culture system of primary neurons and Schwann cells (SCs) to mimic similar symptoms observed in in vivo models of this complication, such as impaired neurite extension and impaired myelination. The model was then utilized to investigate the effects of insulin on enhancing neurite extension and myelination of diabetic neurons. Methods: SCs and primary neurons were cultured under conditions mimicking hyperglycemia prepared by adding glucose to the basal culture medium. In a single culture, the proliferation and maturation of SCs and the neurite extension of neurons were evaluated. In a co-culture, the percentage of myelination of diabetic neurons was investigated. Insulin at different concentrations was supplemented to culture media to examine its effects on neurite extension and myelination. Results: The cells showed similar symptoms observed in in vivo models of this complication. In a single culture, hyperglycemia attenuated the proliferation and maturation of SCs, induced apoptosis, and impaired neurite extension of both sensory and motor neurons. In a co-culture of SCs and neurons, the percentage of myelinated neurites in the hyperglycemia-treated group was significantly lower than that in the control group. This impaired neurite extension and myelination was reversed by the introduction of insulin to the hyperglycemic culture media. Conclusions: Insulin may be a potential candidate for improving diabetic neuropathy. Insulin can function as a neurotrophic factor to support both neurons and SCs. Further research is needed to discover the potential of insulin in improving diabetic neuropathy.

Transgene chgH-rfp expression at developmental stages and reproductive status in marine medaka (Oryzias dancena)

  • Cho, Young Sun;Nam, Yoon Kwon
    • Fisheries and Aquatic Sciences
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    • v.19 no.9
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    • pp.41.1-41.7
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    • 2016
  • Background: The transgenic approach using estrogen-responsive regulator in fish has been given much attention as a potential means to detect and/or address estrogen-related aquatic pollutions. In order to address the development stage- and reproduction status-dependent expression patterns of the chgH-rfp transgene (red fluorescent protein transgene driven by choriogenin H promoter) in marine medaka Oryzias dancena, naturally occurring red fluorescent protein (RFP) signals under non-exposed conditions as well as the transgenically induced RFP signals under estrogen-exposed conditions were assayed. Results: Female transgenics begun to show naturally occurring RFP signals from the age of 7 weeks post hatching (WPH) without experimental estrogen exposure. Afterward, these RFP signals in female transgenics became robust with the progress of ovarian maturation. On the other hand, male transgenics did not show any naturally occurring RFP signal under non-exposed conditions irrespective of developmental stages and maturation statue. Upon exposures using estradiol-$17{\beta}$ (E2) and $17{\alpha}$-ethinylestradiol (EE2), RFP signals were significantly induced specifically in the livers of transgenic males. Conclusions: Male chgH-rfp transgenics were able to keep the "off" state of RFP expression during their entire life cycle unless exposed to exogenous estrogens. Owing to their tight regulation capability of estrogen-responsive transgene, transgenesis of chgH-rfp in male marine medaka could offer a useful model system for future ecotoxicogenomic studies regarding estrogenicity-related issues in aquatic and marine environments.

The Effects of Panax ginseng on TCDD-induced Testicular Atrophy in Guinea Pigs

  • Kim, Wun-Jae;Hwang, Seok-Yeon;Lee, Hyung-Lae;Song, Geun-Song;Kim, Si-Kwan
    • Proceedings of the Ginseng society Conference
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    • 1998.06a
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    • pp.300-311
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    • 1998
  • 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), one of the most notorious toxic environmental pollutants, induces various toxic effects in many organs including testes and is regarded as an endocrine disruptor. Korean ginseng, on the other hand, has been well known for its preventive effects on lox- ins, diabetes melltus and hyperlipidemia. We investigated, histopathologically, the effect of Korean Red ginseng water extract (KR-WE) on guinea pig testes damaged by TCDD. Ninety guinea pigs were divided into 6 groups: normal control (NC) group received vehicle and saline; TCDD,1191kg b.w., was administered intraperitoneally to the single dose TCDD-treated (77) group; 100 mghg b.w.16 and 200mg1kg b.w./d KR-WE were injected intraperitoneally to the preventive groups (PIOO and P2OO, respectively) for 28 days from 1 week before TCDD injection, and to the therapeutic groups (CIOO and C2OO, respectively) for 14 days since 1 week after TCDD administration. Increment of body weight was retarded to a larger extent by TCDD. Moreover, body weight of the 77 group decreased significantly 7 days after TCDD exposure, while that of preventive groups kept increasing. Decrease in body weight was not observed in KR-WE-treated groups. Weight decrease in testes caused by TCDD was remarkably protected by KR-WE. Testicles in 77 group displayed decreased tubular size and maturation arrest at the primary or secondary spermatocyte stage. On the other hand, maturation arrest in germ cells by TCDD was improved in KR-WE treated groups. Almost complete protection of the testes was observed in PIOO and P2OO groups. In addition, the therapeutic effect was noticed in C 100 and C2OO groups. These results provided strong evidence that Korean Red ginseng might be a useful agent for the prevention and treatment of testicular damage induced by environmental pollutants.

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