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マウス味細胞の分化・増殖調節機構におけるインスリンとヒアルロン酸の働き

渡邉, 雄 WATANABE, Yu ワタナベ, ユウ 九州大学

2021.03.24

概要

味蕾に含まれる個々の味細胞は絶え間なくターンオーバーを繰り返しているが、味蕾としての恒常性は常に維持されている。このターンオーバーは、様々なケミカルメディエーターの影響下にあることが予想されるが、その調節メカニズムには不明な点が多い。本研究では、全身のエネルギー代謝において極めて重要な役割を担うホルモンであるインスリンと、細胞外基質の主要な構成成分であり多様な生理活性を持つヒアルロン酸(以下HA)に着目し、その末梢味覚器における働きを探索した。

はじめに、マウス末梢味覚器におけるインスリン関連分子の発現を探索した結果、インスリン受容体は味蕾に広く発現していることわかった。味蕾前駆/幹細胞3次元培養系である味蕾オルガノイドを用いた実験の結果、培地中にインスリンを添加すると、インスリン濃度依存的にオルガノイドコロニーに含まれる味細胞の数、各種味細胞マーカーmRNAの発現量が有意に減少することが明らかとなった。さらに、インスリンシグナリングの下流に存在し、細胞分裂や生存の調節に重要な働きを持つことが知られるmTOR(mechanistic target of rapamycin)は味細胞、またLGR5(leucine-rich repeat-containing G-protein coupled receptor5)陽性の味蕾前駆/幹細胞に発現していた。味蕾オルガノイドで、mTOR経路を薬理学的に阻害すると、コロニー中の味細胞の増加、および各種味細胞マーカーmRNAの発現量増加が見られた。以上の結果は、インスリン-mTOR経路が味細胞の分化・増殖の調節に関わる可能性を示唆する。

次に、HAの末梢味覚器における役割を探索した。マウスの味蕾にはHA受容体であるCd44とHmmr(RHAMM(receptor for hyaluronan-mediated motility))および、HA合成酵素のHas2、Has3(Hyaluronansynthase2、3)とHA分解酵素のHyal1–4(Hyaluronidase1–4)mRNAの発現が見られた。免疫組織化学染色では、CD44が有郭乳頭領域に広範囲に発現しており、HA自体も味蕾内、味蕾周囲に分布していることがわかった。さらに、味蕾オルガノイド培地中にHA合成酵素阻害剤4-methylumbelliferon(4MU)を添加すると、4MU濃度依存的に味蕾オルガノイドのサイズ、および味蕾前駆/幹細胞マーカーであるLgr5、間葉系細胞マーカーであるVim(Vimentin)、HA受容体であるCd44のmRNA発現が有意に減少した。最後に、4MUを40日間継続してマウスに経口投与することにより、オルガノイドを用いた実験同様、各種味細胞マーカーのほか、Lgr5、VimentinのmRNA発現が有意に減少した。さらに、この4MU投与マウスでは味蕾周囲の細胞増殖が劇的に抑制されていることが明らかとなった。以上の結果から、HAは味蕾および周囲組織で合成されており、味細胞に発現するHAの受容体を介して、味蕾前駆/幹細胞の分化・増殖を調節することで味覚器のホメオスタシス維持に関与する可能性が示唆された。

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