2014 Fiscal Year Research-status Report
Regulatory evolution of the turtle shell
| Project/Area Number |
26840121
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| Research Institution | The Institute of Physical and Chemical Research |
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Principal Investigator |
PASCUAL JUAN 独立行政法人理化学研究所, 倉谷形態進化研究室, 研究員 (30594098)
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Keywords | turtle shell / evolution / development / Evo-Devo |
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| Outline of Annual Research Achievements |
As for the first year, I was able to dissect embryos of turtle, chicken and mouse and obtained enough amount of embryonic samples of the carapacial ridge (carapace anlage), limbs and body walls of the turtle, as well as dorsal flanks, limbs and body walls of mouse and chicken embryos as well. Using specific antibodies for different histone modifications, I could obtained the chromatin regions associated with either promoters, active or repressive sequences. The immunoprecipitated DNA was sequenced using high-throughput platforms and the results analysed with different bioinformatics pipelines that have allow me to identify a average of around 5,300 promoters and 39,000 enhancers per turtle tissue. Of those, 494 enhancers are specific to the carapacial anlage.
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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
The research plan proposed stated that the during the first fiscal year I would identify those regulatory elements specific to the carapacial ridge, and not present in mouse and chicken. Although working with non-model organisms, like turtles, usually supposed a research handicap, I have been able to overcome this disadvantage. However, inter-species comparisons are not trivial, are still ongoing. This are estimated to be done by the end of the first quarter of FY2015.
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| Strategy for Future Research Activity |
The plan for this fiscal year is twofold: on the one hand, I will finish the research plan stated in the project application, i.e, reporter assays in amniote embryos. This is the most challenging part of the project, and therefore of uncertain success. On the other hand, and based on the gained experience, I will expand the project to obtain those regulatory elements associated with the transcription factor Lef-1, which is already known to be important for the development of the carapacial anlage. This will allow me drawing a more complete picture of the gene regulatory network active during the carapace formation.
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| Causes of Carryover |
In order to do the bioinformatics analyses smothly, I planned to acquire a powerful computer. However, I have been able to supply this deficit by other means, like the use of high-memory servers available from our laboratory. This has made the project to have superavit at the end of the first fiscal year, and will allow me to expand the project beyond.
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| Expenditure Plan for Carryover Budget |
I plan to expand the sequencing by using other specific antibodies against Lef-1 and beta-catenin, important elements of the Wnt canonical signalling pathway. Both of them are active during the carapace development, and the combination of the histone marks datasets with these newly planned ones will be key to depict a more clear scenario of how the carapace gene regulatory network was shaped during evolution.
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Research Products
(3 results)
