基質金屬蛋白酶在瓢體蟲Aeolosoma viride前端再生扮演的角色

The Roles of Matrix Metalloproteinases in Anterior Regeneration in Aeolosoma viride

曾子倫(Tzu-Lun Tseng)

指導教授 ： 陳俊宏

國立臺灣大學/生命科學院/生命科學系/碩士(2016年)

https://doi.org/10.6342/NTU201601565

摘要

蛋白酶參與在生物體內許多重要的生理機制，像是消化、凝血及組織重組等。而在眾多蛋白酶中，金屬蛋白酶是一群具有最多元酵素功能的蛋白酶，其中基質金屬蛋白酶(Matrix metalloproteinase, MMP)、ADAM及ADAMST為金屬蛋白酶中主要的三個類群，他們都需要金屬離子的協助才能具有正常的酵素活性。而MMP是目前具有最多研究的金屬蛋白酶，他們是主要調控細胞外間質(Extracellular matrix)的分解及構成，已被發現參與在發育及再生過程中的細胞增生及移動。雖然在斑馬魚及渦蟲中MMP已被發現參與再生的過程，但MMP在再生中的機制仍不甚清楚。由於淡水生的環節動物瓢體蟲 (Aeolosoma viride) 具有很強的再生能力，並且使用和脊椎動物類似的機制來進行再生，因此在此篇研究中被用來進行MMP與再生的研究。在此論文的研究結果中，選殖出瓢體蟲中三種基質金屬蛋白酶，其中兩種是鑲嵌在膜上的MMP (MT-MMP)而另外一種是分泌型的MMP。其中兩種MMP在再生早期會高量表現於新生成的組織中，顯示該兩種MMP可能參與在再生的早期過程。進一步去檢測在再生過程中MMP的活性變化，發現到MMP在再生24小時的時候會在新再生組織中具有很高的酵素活性。再以MMP活性的抑制劑GM6001去抑制在再生中MMP的活性，來了解MMP在再生過程中的功能，發現到只有在再生0-12小時內抑制酵素活性會顯著地降低再生的成功率。依據這些結果都確實MMP參與了瓢體蟲的早期頭部再生。

關鍵字

瓢體蟲 ； 再生 ； 基質金屬蛋白酶 ； GM6001

並列摘要

Proteases play important roles in many biological processes. Among these proteases, metalloproteases represent the largest catalytic classes of proteases in many organisms. The three major classes of metalloproteinases are ADAMs (a disintegrin and metalloproteinase domain), ADAMTS (ADAMs with thrombospondin domain), and MMP (matrix metalloproteinase). MMPs play key roles in turnover of extracellular matrix (ECM) and serve as important regulators of cell-ECM interaction during development and regeneration. Although MMPs were found involved in the regeneration in planarian and zebrafish, there is a knowledge gap between planarian and vertebrates. Therefore, the annelid was chosen to fill the gap due to the similarity of regenerative mechanism to vertebrate. In this study, a fresh water annelid with high regenerative ability, Aeolosoma viride, was used to solve this question. In the NGS transcriptome data of A. viride, three MMPs, named as Avi-MMP14, Avi-MMP21 and Avi-MMP17, and one MMP-like gene, Avi-MMP-like gene, were found. Gene expression of Avi-mmp21 and Avi-mmp17 significantly increased at the early stage of anterior regeneration. And, gene expression of Avi-mmp-like gene significantly increased at the late stage of anterior regeneration. However, Avi-mmp14 showed no differences in expression during anterior regeneration. On the other hand, SDS-PAGE gelatin zymography showed the highest protease activity was detected at 24 hours post amputation (hpa) blastema. Furthermore, after treated with GM6001, a MMPs inhibitor, at 0-12 hpa, regeneration was inhibited. These results suggested that MMPs have some effects on early stage of anterior regeneration, and Avi-MMP17 and Avi-MMP21 might be the key factors in this processes. The relationship between MMPs and the regulation of MMPs needs further research to be resolved.

並列關鍵字

Aeolosoma viride ； regeneration ； matrix metalloproteinases ； GM6001

參考文獻

Agata, K., Saito, Y., & Nakajima, E. (2007). Unifying principles of regeneration I: Epimorphosis versus morphallaxis. Dev Growth Differ, 49(2), 73-78.

Agata, K., & Umesono, Y. (2008). Brain regeneration from pluripotent stem cells in planarian. Philos Trans R Soc Lond B Biol Sci, 363(1500), 2071-2078.

Andries, L., Van Hove, I., Moons, L., & De Groef, L. (2016). Matrix Metalloproteinases during Axonal Regeneration, a Multifactorial Role from Start to Finish. Mol Neurobiol.

Apte SS. (2004). A disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motifs: the ADAMTS family. Int J Biochem Cell Biol. 36(6), 981-5.

Armstrong, D. G., & Jude, E. B. (2002). The Role of Matrix Metalloproteinases in Wound Healing. Journal of the American Podiatric Medical Association, 92(1), 12-18.