2020 Fiscal Year Research-status Report
Study of molecular events during scarless body part regeneration in newts
| Project/Area Number |
20K22969
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| Research Institution | University of Tsukuba |
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Principal Investigator |
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| Project Period (FY) |
2020-09-11 – 2022-03-31
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| Keywords | newt / regeneration / scarless / physiology / tissue |
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| Outline of Annual Research Achievements |
To study tissue patterning in the regenerating newt the following transgenic animals were generated. 1 Transgenic Newts expressing the exogenous gene (fluorescent protein) in erythrocytes. To drive exogenous gene expression the mouse GATA1 erythroid enhancer was used. Expression was detected in circulating embryonic erythrocytes. 2 Transgenic Newts expressing the exogenous gene (fluorescent protein) for Shh sonic headhodge cell tracking. Both transgenic animals are now being growth into adult stages.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
We used a previous transgenic protocol to develop the new Gata1 erythroid and Shh cell tracking transgenic system. Transgene plasmids were assemble during the summer 2020, and were available for transgenic microinjection by November 2020, in time for fertilized eggs collection of the newt. All resources for generation of transgenic animals for this were available, but some purchased items (reagents) were delayed due to impact of COVID on shipping.
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| Strategy for Future Research Activity |
GATA1 erythroid tracking animals will be validated for expression after metamorphosis. Also tissue and blood samples must be anazlyed with immunohistochemistry. After validation newtic1 shRNAi will used with this system to knockdown newtic1 in eyrthrocytes during limb regeneration. Shh tracking animals will be validated for expression after metamorphosis. Also tissue and blood samples must be anazlyed with immunohistochemistry using anti-shh antibody.
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| Causes of Carryover |
The reason for 44円 to be used for next fiscal year was due to research delay to order plastic cups for rearing GATA1 newt larvae. However research plan will contain because newt larvae are still developing and additional plastics cups are needed. Therefore, 44円 can be used for same research purpose.
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