2021 Fiscal Year Research-status Report
Investigation of the role of radial glia in regulating LysoPtdGlc required for nervous system development
| Project/Area Number |
20K06884
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| Research Institution | Kyoto University |
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Principal Investigator |
GUY ADAM・TSUDA 京都大学, 生命科学研究科, 准教授 (30634779)
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| Project Period (FY) |
2020-04-01 – 2023-03-31
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| Keywords | Lipid biology / Neurochemistry / Spinal cord / Lysophospholipid |
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| Outline of Annual Research Achievements |
Despite most neural cells expressing a complex mixture of different isoforms of secreted phospholipase A2 (sPLA2) enzyme, I previously discovered that radial glia in developing mouse spinal cord specifically express one isoform: sPLA2 V (gene name pla2g5). In FY2021, I found that mice genetically lacking sPLA2 V developed an abnormally large TrkA domain in spinal cord, that was statistically significant. Therefore I concluded that radial glial sPLA2 V plays a role in axon guidance and nociceptive axon tract formation. According to my hypothesis that this should be specific to nociceptive axons, I carried out antibody staining of TrkC (a marker specific to proprioceptive axons, the second major sensory input from the periphery) in the spinal cords of sPLA2 V knockout mice and wildtype siblings. TrkC domain did not show a statistically significant difference in size compared to wildtype, strongly suggesting that the phenotype observed in nociceptive axons in sPLA2 V KO mice was specific.
This FY I established a collaboration with Juntendo University, to perform QTRAP LC-MS/MS tandem mass spectrometry in order to measure the concentration of endogenous LysoPtdGlc in chick and mouse embryonic spinal cord tissue. We were able to successfully detect LysoPtdGlc in E8 chick and E14 mouse wildtype embryonic spinal cord, but have yet to quantify it (using internal standards of known molarity).
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
My ability to conduct research has been disrupted by:
Change of institution (I left Riken in April 2021 to move to a faculty position at Kyoto University).
Effects of coronavirus pandemic precautions, including overseas graduate student unable to enter Japan due to Japanese government travel restrictions.
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| Strategy for Future Research Activity |
In FY2022 I will complete the measurement of concentration of LysoPtdGlc in the developing spinal cords of mice genetically lacking sPLA2 V (pla2g5 knockout mice) compared to their wildtype siblings. According to my hypothesis, radial glial sPLA2 V is required for the production of LysoPtdGlc, which is the axon guidance cue that mediates the correct patterning of TrkA-expressing nociceptive afferents during spinal cord development. I expect that knockout mice will have a much lower concentration of LysoPtdGlc, when compared to wildtype.
I have submitted an abstract, which has been accepted, for presentation as a psoter at the 2022 JNSS conference.
I am currently preparing a manuscript to publish these results and I aim to submit to an international peer-reviewed journal during FY2022 once the mass spectrometry data are ready.
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| Causes of Carryover |
I was unable to fully carry out research activities due to coronavirus restrictions on entry to Japan by overseas students. Also I participated in a scientific conference as virtual presenter online only, which required no travel expenses. Approximately 160,000 of budget was not spent. In FY2022 I will spend this full amount on consumables, and depending on pandemic conditions, on travel to attend scientific conferences to present my research results.
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Research Products
(2 results)
