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研究発表を行った学会；
・第４７回日本発生生物学会年会
　２０１４年５月２７日〜３０日（名古屋）
タイトル；Akhirin is involved in the neural stem cell regulation in the mouse spinal cord.
発表者；Athary Felemban　氏
　　　（熊本大学　大学院生命科学研究部　神経分化学分野）
Abstract；
In our lab we identified a secreted molecule called Akhirin (AKH), which consists of one LCCL (Limulus factor C, Coch-5b2, and Lgl1) domain and two von Willebrand factor domains. AKH displays high structural homology to vitrin and cochlin and has heterophilic cell adhesion activity. To understand the functional analysis of AKH we performed in situ hybridization to check the expression of AKH at different developmental stages of chick and mouse embryos. AKH was strongly expressed around the central canal and floor plate in chick spinal cord at stage 28. In mouse case, a strong signal was observed also around the central canal of the spinal cord at E13. Interestingly, the signal was restricted around the central canal up until the adult stage where the location was known to contain neural stem cells. To determine whether AKH is involved in an important role for prolifereation and differentiation of a mouse spinal cord in knockout mice; BrdU was injected into time-pregnant mice at E10 and E16. We observed that the number of cell proliferations had significant decreased in AKH -/- compared with WT mice. Furthermore, in the developing transgenic Nestin-GFP embryos, WT mice Nestin-GFP were expressed stronger than Nestin-GFP in AKH -/- mice around the central canal of the spinal cord at P10. To examine whether AKH has a role in neural stem cells regulation in mouse spinal cord, neurosphere culture was performed from littermate WT and AKH -/- mice using the spinal cord sections at p6. After five days in vitro culture, we can observe the neurosphere in WT and AKH -/-, interestingly; the diameter of WT neurosphere was significant bigger then AKH -/- neurosphere. Immunohistochemistry using different markers on secondary neurospheres, such as: Vimentin, Cyclin D2, and Ki67. We observed the layer formation in WT neurosphere, but we did not detected in AKH -/-. Finally, we checked the expression pattern of AKH in the intact and injured spinal cord, and we found that an AKH signal was up regulated in injured adult spinal cord. Our conclusion is that Akhirin is involved in the neural stem cell of a mouse spinal.