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Hur, Sung-Pyo,Lim, Bong-Soo,Hwang, In-Joon,Kim, Se-Jae,Ryu, Yong-Woon,Hur, Sang-Woo,Song, Young-Bo,Jeong, Hyung-Bok,Baek, Hae-Ja,Takemura, Akihiro,Lee, Young-Don The Korean Society for Integrative Biology 2012 Animal cells and systems Vol.16 No.2
We investigated the effects of fadrozol, an aromatase inhibitor (AI), and $17{\alpha}$-methyltestosterone (MT) on the induction of sex change in juvenile longtooth grouper $Epinephelus$ $bruneus$, via histological observation of gonads. Changes in the mRNA expression of GtH subunits (FSH-${\beta}$ and LH-${\beta}$) in the pituitary, and estradiol-$17{\beta}$ (E2) and 11-ketotestosterone (11-KT) levels in the blood were also surveyed after AI and MT treatment. Juvenile longtooth groupers ($113{\pm}17g\;body\;weight$; $16.2{\pm}1.2cm\;body\;length$) received intramuscular injections of AI at 3 (3-AI) and 5 (5-AI) mg/kg BWdoses and MT at a 5 mg/kg BW (5-MT) dose. At week 7 post-injection, 3-AI and 5-MT oocytes were degenerated, and gonads of the 5-AI group initiated spermatogenesis. At week 21 post-injection, 3-AI- and 5-MT-treated gonads contained spermatogonia and spermatocytes, while 5-AI treatment induced advanced stages of spermatogenesis. The serum E2 level showed no significant differences throughout the experimental period, whereas that of 11-KT was significantly elevated in the 5-AI group at weeks 7 and 21 post-injection. A significant increase in the expression of FSH-${\beta}$ mRNA was evident in the 5-AI group at week 21 post-injection. In contrast, LH-${\beta}$ mRNA expression did not significantly differ among groups during the experimental period. These results imply that sex change has two stages in the longtooth grouper. In the first stage, oocytes are degenerated by the stimulation by 11-KT, and in the second stage spermatogenesis occurs, owing to the co-effects of 11-KT and FSH-${\beta}$.
Sang-Woo Hur,Chi-Hoon Lee,Hea-Ja Baek,Choong-Hwan Noh,Sang-Hyun Han,Seung-Bo Oh,Ji-Sung Moon,Young-Don Lee 한국발생생물학회 2014 한국발생생물학회 학술발표대회 Vol.2014 No.9
As a preliminary investigation into the effect of environmental factors control for gonadal development, we examined the involvement of photoperiod and water temperature in ovarian development of Epinephelus. akaara. For the induction of sexual maturation, E. akaara reared in recirculating aquaculture system (RAS). During November 2013, the photoperiod and water temperature was adjusted to 12L:12D and 18℃, respectively. In the photo-thermal treatment group, every 3 weeks daylight was increased as follows a 13L:11D and 14L:10D, and control group was maintained under natural condition. After 9 weeks, water temperature was increased 23℃ in photo-thermal treatment group. The sampled fish every 3 weeks revealed increase in gonadosomatic index (GSI; 5.18±1.38), oocyte diameter and vitellogenic oocytes (423.9±36.1 ㎛) were observed in gonads 12 weeks under photo-thermal treatment group. However, ovarian development was maintained immature stage in control group. In this environmental factors manipulation trial, seventy one of the 95 females (578.4 ± 25.4 g in mean body weight, 31.0 ± 0.5 cm mean total length) treated with HCG injection (doses 500 IU/kg BW) were induced ovulation by artificial stripping. The total volume of ovulated eggs were 3,470 ml and the total volume of fertilized eggs was 3,295 ml. The fertilization rate and hatching rate were 95% and 98%, respectively. These results suggest that the photoperiod as well as water temperature are major environmental factors in triggering the gonadal development of E. akaara.
( Sung Woo Kim ),( Won Hee Hur ),( Kwang Soo Lyoo ),( Jung Eun Choi ),( Sung Woo Hong ),( Young Kee Lee ),( Seung Kew Yoon ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1
Background: NAFLD (nonalcoholic fatty liver disease) has a wide spectrum of liver pathogenesis from simple hepatic steatosis to cirrhosis. NASH (Nonalcoholic Steatohepatitis) is characterized by hepatocyte injury and inflammatory cell infiltration, which is linked with peripheral insulin resistance and increased triglyceride in the liver. Pharmacological properties of oleuropein are anti-oxidant, anti-inflammatory, and anti-ageing. The purposes of this study was to establish an NASH mouse model fed with HFD and to demonstrate the pharmacological effect of oleuropein using the appropriative animal model. Methods: C57BL/6 mice were divided into 3 groups; a normal diet group (ND), a 60% high fat diet group (HFD), 0.05% oleuropein-supplemented high fat diet group (HWO). HWO were fed 0.05% oleuropein-supplemented high fat diet after 6 months. The effect of oleuropein on these models was studied using biochemical, histological and molecular markers. Expression of mRNA level (related adipogenesis, inflammation, and fibrosis) was analyzed by real-time PCR between HFD and HWO. Results: The body weight, total cholesterol, TG, FFA, AST, and ALT values of HFD for 6, 9, and 12 months were higher than that of ND. The NAFLD activity score (NAS) of HFD were increased to inflammation stage at 6 month, compared to ND. HWO was significantly increased to NAS and fibrosis grade, but steatosis grade was not changed. The mRNA levels of LXR, TNF-α and collagen was decreased with treatment period in HWO, but ap2 and α-SMA was not changed. Conclusions: We produced a high fat diet induced NASH model that displayed histopathological features of NAFLD to NASH. The therapeutic effect of oleuropein was prevented with progression of NAFLD to NASH. Therefore, it is speculated that oleuropein may be pharmacologically useful to prevent the progression of steatohepatitis and fibrosis and could be a promising agent of medicine for human NASH. This study was supported by a grant of the Korean Health Technology R&D Project, Ministry of Health & Welfare, Republic of Korea. (A090282).
( Sung Woo Hong ),( Won Hee Hur ),( Jung Eun Choi ),( Young Ki Lee ),( Seung Kew Yoon ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1
Background: Recent studies have described that cancer stem cell plays a key role in radioresistance. The cell surface marker CD133 has been known recently as a cancer stem cell marker expressed in HCC. Recently, ADAM17 was reported to contribute to metastasis. The aim of this study is to investigate the molecular and cellular mechanisms of liver cancer stem cell and ADAM17 in metastasis after irradiation in HCC. Methods: Huh-7CD133+ and Huh-7CD133- sorted cells were exposed to g-irradiation. We investigated the key gene/pathway responsible for metastasis in post-irradiated liver cancer stem cells, CD133+/- cells using cDNA microarray. Metastatic activity was analyzed by cell migration assay. Also the expressions of the MMP-2 and MMP-9 were measured by gelatin zymography. We confirmed ADAM17 gene expression on time dependent by real-time PCR, western blot in sorted cells. ADAM17 is suppressed effectively using ADAM17 lenti virus shRNA in Huh-7 cell line. CD133+/- cells that were suppressed ADAM17 gene expression were analyzed migration activity. Results: After cDNA microarray analysis, eighty nine metastasis related genes were upregulated. Especially we found that the ADAM17 gene was more increased in CD133+ cells than CD133- cells treated with g-irradiation. In addition CD133+ cells from radiation exposure were consistently expressed higher levels of vascular endothelial growth factor by Multiplex cytokine analysis. After irradiation, CD133+ cells migrated more actively, and showed an increased invasion rate compared to CD133- cell. Gelatin zymography showed that MMP-2, -9 expressions are significantly higher in CD133+ cells. Furthermore, shADAM17CD133+ cells reduced migration activity after g-irradiation exposure. Conclusions: These results suggest that CD133+ cells have more metastatic capacity than CD133- cells after treating irradiation. These data further suggest that molecular therapeutic inhibition of cell migration and invasion through ADAM17 suppression may represent a new approach for improving the therapeutic efficacy of radiotherapy for HCC. This work was funded by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (No. 2011-0027071).