2019 Fiscal Year Research-status Report
魚類の給餌を最適化する新たなアプローチ：オートファジーによる生体防御機構の解明
| Project/Area Number |
18K14520
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| Research Institution | Ehime University |
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Principal Investigator |
Mohapatra Sipra 愛媛大学, Kyushu University, Researcher (80715441)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Keywords | Autophagy / Hexokinase / estrogen / Liver / Fish / estrogen receptor |
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| Outline of Annual Research Achievements |
I have characterized the HKII, III and IV profiles in Red Sea bream liver and confirmed their roles in liver autophagy regulation. Fasting and E. trade challenge experiment using both medaka and Red Sea bream, showed that several autophagy gene were significantly altered. I also isolated a 5KB promoter sequence of HKII from both fish species and found that, 17b-estradiol significantly increased the HKII promoter activity in vitro. Later, using two Estrogen receptor Knockout medaka line and CHIP analysis it was confirmed that HKII is a direct target of estrogen receptor. These data cumulatively suggests that, HKII is a prime candidate of sex biased liver autophagy in fish and can be a potential marker for ecofriendly aquaculture in future.
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
Earlier, I planned to isolate HKII, HKIII and HKIV from Red Sea bream and analyze their tissue distribution and cellular localization. I also planned to establish primary hepatocyte culture method, perform serum and glucose starvation under various hexokinase over expression and knockodown condition in vitro, conduct promoter analysis invitro using mammalian cell line. comparative invivo starvation and infection experiment using medaka and Red Sea bream were also planned. I have successfully finished all the planned experiment at least three times and analyzed the data.
Additionally I performed indepth analysis on sex biased autophagy and estrogen regulation and published a peer reviewed research article. I have also generated HKII knockout medaka which will be helpful to analyze the downstream cascade of HKII regulation in fish,
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| Strategy for Future Research Activity |
I will decipher the downstream genes of HKII pathway using chromatin immunoprecipitation and other relevant analysis both invitro and invivo. I will also look into the hepatocyte mitochondria and get an idea about the mitophagy and HKII association. If necessary, I will investigate several other autophagy related candidates to confirm the prospects of autophagy in aquaculture management.
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| Causes of Carryover |
During the initial period (since November 2018- May 2019, I was on maternity and child care leave, which affected my timely execution of Planned research. Hence I got delayed by few months. I will use the money in this fiscal year
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