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J. Hefei Univ. Tech. (Nat. Sci.)

合肥工业大学学报(自然科学版)

JOURNAL OF HEFEI UNIVERSITY OF TECHNOLOGY (NATURAL SCIENCE)

Competent Authorities: Ministry of Education of the People's Republic of China

Sponsor by: Hefei University of Technology

ISSN 1000-1646, CN 34-1001/N

Using cpl-1(qx304); MAH215 transgenic Caenorhabditis elegans to investigate the role of CPL-1 in aging-related autophagy

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利用 cpl-1(qx304); MAH215 转基因线虫研究 CPL-1 在自噬中的作用

Journal Information

J. Hefei Univ. Tech. (Nat. Sci.), Dec.2025, Vol.48 No.12: 1652-1656, 1677

DOI: 10.3969/j.issn.1003-5060.2025.12.011

Authors

ZHOU Ying, LIU Jian, LIN Yan

(School of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, China)

Abstract and Keywords

Abstract: Caenorhabditis elegans is a widely used aging model, in which lysosome-mediated autophagy is gradually reduced with aging. However, the role of the lysosomal cathepsin L-like protease (CPL-1), a critical lysosomal protease, in the regulation of aging remains unclear. In order to explore the role of CPL-1 in aging process, the cpl-1 deleted worm cpl-1(qx304) was used to cross with the transgenic nematode MAH215(sq I s II [lgg-1p::mCherry::gfp::lgg-1+rol-6]), a dual-labeled autophagy substrate (LGG-1) with red and green fluorescence (mCherry::GFP::LGG-1). Through the fluorescence expression determination and DNA sequence analysis, it was confirmed that the transgenic worm cpl-1(qx304); MAH215 was successfully constructed with the expression of mCherry::GFP::LGG-1 and deficiency of cpl-1. Furthermore, the expression of mCherry::GFP::LGG-1 was explored, and it was found that the yellow fluorescence (red and green fluorescence co-localization) expression in wild-type MAH215 worms was significantly increased during aging process, and further increased in cpl-1 deleted worms. The research revealed that the autophagy substrate LGG-1 was

Keywords: Caenorhabditis elegans; autophagy; aging; cathepsin L-like protease(CPL-1); cross

作者信息

周颖，刘健，林燕

（合肥工业大学食品与生物工程学院，安徽合肥230601）

摘要和关键词

摘要: 秀丽隐杆线虫是一种被广泛使用的衰老模型,其溶酶体介导的自噬水平随衰老进程逐渐下降;溶酶体组织蛋白酶L样蛋白(cathepsin L-like protease, CPL-1)作为溶酶体中的一种关键组织蛋白酶,其在衰老调控中的作用尚不明确。为了研究CPL-1在衰老过程中的作用,文章利用cpl-1缺失线虫cpl-1(qx304)与自噬底物LGG-1双荧光(绿色和红色)标记的转基因线虫MAH215(sqI sⅡ[lgg-1p;:mCherry;::gfp;::lgg-1+rol-6])进行杂交,通过荧光监测线虫表型,再对其进行DNA测序分析,筛选出成功表达mCherry:GFP::LGG-1的cpl-1缺失线虫cpl-1(qx304);MAH215。通过荧光表达监测,发现随着衰老的进程,野生型MAH215线虫中LGG-1的黄色荧光(红色和绿色荧光共定位)表达显著增加,而cpl-1缺失进一步增加LGG-1的黄色荧光水平。研究结果表明:在衰老过程中,自噬水平降低,自噬底物LGG-1逐步聚集;而cpl-1缺失则进一步增加LGG-1的聚集,加速了衰老的进程。因此,通过构建的cpl-1(qx304);MAH215转基因线虫确定了CPL-1在衰老相关自噬调控中的重要作用。

关键词: 秀丽隐杆线虫；自噬；衰老；组织蛋白酶L样蛋白(CPL-1)；杂交

Funding

国家自然科学基金资助项目（32070757）；中央高校基本科研业务费专项资金资助项目（JZ2022HGQA0236）

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