Mechanisms that regulate the number of cells that constitute the body have remained largely elusive. We approached this issue in the ascidian, Halocynthia roretzi, which develops into tadpole larva with small number of cells. Embryonic cells divide 11 times on average from fertilization to hatching. The number of cell division rounds varies between tissue types. For example, notochord cells divide 9 times and give rise to large postmitotic cells in the tadpole. The number of cell division rounds in the partial embryos that were derived from tissue-precursor blastomeres isolated at the 64-cell stage also varied between tissues, and coincided with their counterparts in the intact whole embryos to some extent, suggesting tissueautonomous regulation of cell division. Manipulation of cell fates in notochord, nerve cord, muscle, and mesenchyme lineage cells by inhibition or ectopic activation of the inductive FGF signal changed the number of cell division according to the altered fate. Knockdown and missexpression of Brachyury (Bra), an FGF-induced notochord-specific key transcription factor for notochord differentiation, indicated that Bra is responsible not only for notochord differentiation but also regulates the number of cell division rounds in the notochord lineage cells, suggesting that Bra activates a putative machinery to stop cell division at the specific stage. Results of precocious expression of Bra suggested that the machinery refers the developmental clock that is likely shared in other blastomeres than notochord, and functions to terminate cell division at three rounds after the 64-cell stage. Bra does nothing about the progression of developmental clock itself.

The transcription factors, DMRT1 and FOXL2, play a role in fish sex differentiation of the bipotential precursor into the male and female pathway, respectively. In order to provide the molecular background for understanding hormonal regulation in sexual determination and differentiation in the red-spotted grouper, Epinephelus akaara, one of commercially important epinephelines, and is often used to study protogynous sex change. First, we amplified the partial sequence of two genes (DMRT1 and FOXL2) from the gonad of red-spotted grouper. Also, we surveyed the tissue-specific and sex-specific expression pattern of each genes by RT-PCR. The effects of 17α-methyltestosterone (MT) in the sexually immature gonad of red-spotted grouper were investigated by Real-time quantitative RT-PCR. Fish were treated with MT-dipping method from around 70 days after hatching (DAH) for two month. DMRT1 and FOXL2 cDNA flagments consist of 489 and 836 base pairs (bp) and encodes a protein of 162 and 278 amino acids, respectively. RT-PCR revealed that DMRT1 mRNA was expressed higher level in the testis. Foxl2 was expressed extensively in the neural and peripheral tissues with its highest level in the ovary, indicating a potential role for Foxl2 in the brain-pituitary-gonad axis. Real-time quantitative RT-PCR analyses showed that DMRT1 mRNA expression was upregulated in the MT-treated fish. These results suggest that the sex inversion of red-spotted grouper by MT might be due to the suppression of FOXL2 gene expression, and resulting in the induction of the 11-KT secretion.

Controllable transgenic expression systems in transgenic animal model are valuable to the development of therapeutic approaches in human medical fields. The aim of this study was to 1) produce a transgenic cloned dog using inducible tetracycline vector system, and 2) investigate whether the transgenic cloned dog could be induced the transgene expression using doxycycline (Doxy). Canine fetal fibroblasts were infected with retroviral vectors designed to express the enhanced green fluorescent protein (eGFP) gene under the control of tetracycline-inducible promoter. For somatic cell nuclear transfer (SCNT), nucleus of an in vivo matured oocyte was removed and an eGFP expressed cell cultured with 1 ㎍/㎖ of Doxy was injected. After electrical fusion and chemical activation, the reconstructed embryos were transferred to a recipient and pregnancy diagnosis was performed by ultrasonography. Experiment I evaluated the mean fluorescence intensity (MFI) of infected cells while the cells were cultured in the presence of 1 ㎍/㎖ of Doxy for 5 days, and then in the absence of Doxy for 7 days using fluorescence-activated cell sorter. Experiment II was designed to produce an eGFP controllable transgenic cloned dog via SCNT. For verification of transgenic dog, experiment III was performed Southern Blot analysis and observation in vivo regulation of eGFP expression in the cloned dog treated with 100 ㎎/㎏ of Doxy every 2 days for 2 weeks under ultraviolet light. In experiment IV, western blot was used to detect eGFP increase and decrease in skin tissues of transgenic dog under the presence or absence of Doxy. In the results of experiment I, the MFI for infected cells was rapidly increased to approximately 42.3 times after 3 day-treatment compared to pre-treatment and quickly decreased 3 days after ceasing the treatment. In experiment II, a total of 203 embryos were transferred to nine recipients and three pregnant delivered three pups (Tet-on eGFP 0, Tet-on eGFP 1, and Tet-on eGFP 2) by C-sec and Tet-on eGFP 2 among them is still alive. All cloned pups were genetically identical to the donor cell. Tet-on eGFP 2 showed an apparent in vivo eGFP expression on her body after Doxy administration in experiment III. The result of Sothern blotting showed that the transgene insertion was detected from the three cloned dogs and all organs of Tet-on eGFP 1. Experiment IV indicated that a robust eGFP expression in skin tissue of Tet-on eGFP 2 rapidly increased after Doxy treatment and gradually decreased to basal level on 9 weeks after ceasing the treatment. In conclusion, we report here for the first time an inducible transgenic system in canine species and it can stably induce the transgene expression at intended time. This study has demonstrated the capacity to generate transgenic model dog which could regulate the transgene and it would contribute to human medical research fields.

The objective of this study was to investigate the effects of oxygen tension during in vitro maturation of porcine oocytes on the nuclear maturation and differences in gene expression. Cumulus-oocyte complexes (COCs) were collected from ovaries obtained at a local slaughterhouse, matured for 44 hours in TCM199 supplemented with porcine follicular fluid (pFF) under 5% or 20% oxygen concentration. In results, oxygen tension had no significant effects on nuclear maturation. Relative poly(A) mRNA abundance of MnSOD, CCNB1, LDHA, G6PD, BCL, GPX1, IGFR2, GLUT1, BAX, GREM, PTGS2 was analysed in cumulus cells. GLUT1, G6PD, LDHA were up-regulated in the cumulus cells matured in low oxygen, suggesting a higher glucose uptake and an increase in anaerobic glycolysis, whereas CCNB, MnSOD were up-regulated in the cumulus cells matured in high oxygen, which suggest a higher activity of mitosis-promoting factor and antioxidant response. In conclusion, cumulus cells increase in glucose metabolism via anaerobic glycolysis under low oxygen concentration and show significant change in antioxidant against oxidant damage or apoptotic response under high oxygen concentration. For such an effect of cumulus cells, oocytes could be matured normally regardless of various oxygen concentration.

Obesity is often associated with an impairment of the hypothalamic-pituitary-gonadal axis and also unites with reproductive defects and reduced fertility. The leptin-deficient ob/ob mouse is characterized by a morbid obesity and infertility. In the present study, we examined when ob/ob mouse lost their fertility activity according to aged dependent were investigated. The major organ systems in the body, the ovaries of females are the first to exhibit impaired function with advancing age. Here, ob/ob and ob lean various (4, 6, 9, 12 and 24 wks) aged mice were used. Histology (H & E staining) of ovary, vaginal smear and Reverse transcription polymerase chain reaction (RT-PCR) analysis were carried out. Hematoxylin/eosinstained sections show normal histology in ob lean type mice characterized by the presence of multiple primary and secondary follicles and well developed corpora lutea (CL). Ovaries from ob/ob mice, showed 4 and 6 wks mice similar to ob lean type but according to aged dependent ob/ob mice few primary and secondary follicles was seen. Many large empty follicles were present and no proper developed CL, especially 24 wks mice. Vaginal smear showed abnormal estrous cycle in ob/ob mice. Ovaries of ob/ob mice, mRNA results showed that ovulated gene TIMP1, Pgsh2 and Lhcgr expression levels are decreased according to aged dependent compared to ob lean type mice. These data suggest that aged dependent specifically when 24 wks of ob/ob mice lost their reproductive function.

In the 1910s and 1920s, T. S. Eliot had frequently made interesting remarks on politics and commented on political issues through literary journalism. In his writings, he expressed a strong dislike for liberal democracy, a main current of British as well as universal political movements then. After taking up the editorship of The Criterion in 1922, Eliot advanced his opinions persistently against liberal democracy on political and literary issues through the quarterly review.This paper examines Eliot’s political ideas reflected in his various writings in The Criterion right after his conversion to the National Church of England in 1927. This examination finds that the political ideas of Eliot and other contributors in the review have close affinities with those expounded by the French political thinker, Charles Maurras, who was a champion of French monarchism, Classicism, Catholicism, and who condemned liberal democracy after the French Revolution. Although Eliot retained some of his family politics, much of his reactionary ideas against liberal democracy germinates from the thoughts of Maurras. For Eliot, liberal democracy in politics was not conducive to the excellence in the of arts as well as modern culture. However, after the condemnation of Maurras by the Vatican, he turns to Christian politics, which must be in service of the Church.

강섬유를 혼입한 콘크리트(Steel Fiber Reinforced Concrete, SFRC) 보는 강섬유의 우수한 인장강도로 인하여 일반 철근콘크리트 보에 비하여 높은 전단강도를 가진다. 이 연구에서는 강섬유 혼입율에 따른 SFRC 보의 전단거동을 규명하기 위하여 실험을 수행하였으며, 특히, 압축영역에서의 비균열 콘크리트 단면의 전단저항 분담율을 분석하였다. 또한, 이 연구의 실험결과 및 기존에 보고된 87개의 실험 데이터를 수집하여 SFRC보의 전단예측식들에 대한 정확도를 평가하였다. 강섬유의 혼입율이 증가할수록 전단강도는 증가하는 경향성을 나타내었다. 그러나, 강섬유 혼입율이 0.5%인 실험체는 사인장 균열 이후 갑작스럽게 파괴되었고, 강섬유의 혼입율이 2.0%인 실험체에서는 전단보강효율이 감소하는 것으로 관찰되어 최대 전단보강효율을 가질 수 있는 혼입율은 1~2% 사이에 있을 것으로 추정된다. 또한, 압축영역에서의 비균열 콘크리트 단면의 전단저항 분담율은 약 21% 이상으로 관찰되었으며, 이 연구에서 평가된 SFRC보의 전단강도에 대한 기존 제안식 중에서 오영훈 등이 제안한 식이 비교적 정확하게 전단강도를 예측하였다.