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Biology subjects

Ihara, N.

Publications and source records attributed to Ihara, N..

2 recordsLinked to original sources

Auxin-mediated rapid degradation of selective proteins in hippocampal neurons

Genetic manipulation of protein levels is a promising approach to identify the function of a specific protein in living organisms. Previous studies demonstrated that the auxin-inducible degron (AID) strategy provides rapid and reversible degradation of various proteins in fungi and mammalian mitotic cells. In this study, we employed this technology to postmitotic neurons to address whether the AID system could be applied to the nervous system. Using adeno-associated viruses, we simultaneously introduced EGFP fused with an AID tag, and an F-box family protein, TIR1 from Oryza sativa (OsTIR1) into hippocampal neurons. In dissociated hippocampal neurons, EGFP fluorescence signals rapidly decreased when adding a plant hormone, auxin. Further, auxin-induced EGFP degradation was also observed in hippocampal acute slices. Taken together, these results open the door for neuroscientists to manipulate protein expression levels by the AID-system in a temporally-controlled manner.

neuroscience↗

Fetal therapy model of myelomeningocele with three-dimensional skin using amniotic fluid cell-derived induced pluripotent stem cells

Myelomeningocele (MMC) is a congenital disease without genetic abnormalities. Neurological symptoms are irreversibly impaired after birth. No effective treatment has been reported to date. Only surgical repairs have reported so far. In this study, we performed antenatal treatment of MMC with an artificial skin using induced pluripotent stem cells (iPSCs) generated from a patient with Down syndrome (AF-T21-iPSCs) and twin-twin transfusion syndrome (AF-TTTS-iPSCs) to a rat model. We manufactured three-dimensional skin with epidermis generated from keratinocytes derived from AF-T21-iPSCs and AF-TTTS-iPSCs and dermis of human fibroblasts and collagen type I. For generation of epidermis, we developed a novel protocol using Y-27632 and epidermal growth factor. The artificial skin was successfully covered over MMC defect sites during pregnancy, implying a possible antenatal surgical treatment with iPSC technology.

cell biology↗