| 研究実績の概要 |
I have examined whether the adhesion properties of neurons regulate columnar unit formation as suggested by Differential Adhesion Hypothesis by using a fly brain as a model. 1)I have visualized the subsets of columnar neuron (R7, R8, L1-5, T1, Mi1, Tm1, Tm2, Dm4, Pm4-like neurons, etc.) as well as the expression and localization of Ncad during column formation by using fly strains expressing GFP under the control of single neuron type-specific Gal4 and GFP fused Ncad respectively. 2)To examine Ncad and Fmi loss of function effect I have generated a visual center specific mutant cell clones of Ncad and Fmi by somatic recombination technique. Specific recombination constructs, generated in our laboratory, were used in this experiment. I have examined the column shape, size and position of R-axons within the column at larval and pupal stages using Ncad antibody staining as well as specific Gal4 drivers that could visualize the column shape by GFP expression. Brain samples were examined by using a laser scanning confocal microscopy system. I have also performed the loss and gain of function of Ncad and Fmi in a subset of columnar neurons expressing UAS-NcadRNAi/fmiRNAi or UAS-Ncad/fmi constructs of different strength under the control of single type neuron-specific Gal4 drivers. The morphology of columns and phenotypic changes in neuron projection were analyzed in larvae, papa and adult flies by confocal microscope.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
Currently I have finished my scheduled experiments as described above for the previous academic year and writing a manuscript to publish the results. My preliminary results confirm my working hypothesis and show that differential adhesion among different types of neuron within the columns is necessary for the proper column establishment. Obtained results were presented at the 40th Annual Meeting of Japan Neuroscience Society, July 20-23, 2017, Tokyo, Japan, and the 49th Annual Drosophila Conference, April 11-15, 2018, Philadelphia, USA. My manuscript has been sent to the language editing service and is expected to be submitted by the end of this month.
Currently I'm continuing experimental work and examining the interaction of different columnar neurons within the column in wild type flies and in loss of or gain of function conditions for Ncad in single type of columnar neurons.
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| 今後の研究の推進方策 |
First of all, I will submit the manuscript to get it published.
During this academic year, I plan to examine the inter-neuron interaction between columnar neurons that corresponds to one column using single type neuron-specific Gal4 and LexA constructs,generated in our laboratory and recombined with UAS-GFP and LexAop-RFP construct, that allows me to visualize two different subsets of neurons within the column at the same time. I will perform Ncad and Fmi knock down or gain of function experiments in the subsets of neurons of one type and observe its influence on the projection pattern and localization within columns of different types of neurons. At the beginning, I plan to focus on four types of columnar neurons R7, R8, Mi1 and L1-5 and then expand my research on other types of columnar neurons. At the same time, I plan to perform the screening of the role of other Cadherin family members like Ncad2 in the column formation. RNAi knock down experiments will be performed to manipulate the level of other cadherins in a single type of columnar neurons.
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