Ishan Mohamed, Skipper Isabell R, Rozario Tania
Department of Genetics and Center for Tropical & Emerging Global Diseases, University of Georgia, Athens, GA, United States.
Department of Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, GA, United States.
Biol Methods Protoc. 2025 Feb 13;10(1):bpaf011. doi: 10.1093/biomethods/bpaf011. eCollection 2025.
is a parasitic tapeworm that utilizes rats as hosts and offers advantages over human parasitic tapeworms and free-living flatworms as a model system to study the biology and pathology of helminth infections. is minimally infectious to humans, easy to maintain in the lab, demonstrates impressive growth, regeneration, and reproductive capabilities, and is amenable to loss-of-function manipulations. As an emerging model, tool development is critical to increasing the utility of this system. This study introduces a novel protocol for that combines fluorescent in situ hybridization (FISH) and 2'-Deoxy-2'-fluoro-5-ethynyluridine (F--EdU) uptake and staining. Our protocol allows for the spatial detection of gene expression and simultaneous identification of proliferating cells. Dual labeling of F--EdU and stem cell markers revealed a distinct expression pattern in different anatomical regions, especially in the head and neck. We demonstrate optimal labeling without permeabilization, streamlining the protocol. We also demonstrate generalizability using FISH for other tissue markers. The protocol was applied to perform bulk lineage tracing, revealing that stem cells can differentiate into neuronal and tegumental cells within 3 days. Our protocol provides an important tool in the arsenal for investigating gene expression and cell proliferation in , contributing valuable insights into the biology of parasitic tapeworms and potentially opening new avenues for the study of human parasitic tapeworms.
是一种寄生绦虫，以大鼠为宿主，作为研究蠕虫感染生物学和病理学的模型系统，它比人类寄生绦虫和自由生活的扁虫具有优势。它对人类的感染性极小，易于在实验室中饲养，具有令人印象深刻的生长、再生和繁殖能力，并且适合进行功能丧失操作。作为一种新兴模型，工具开发对于提高该系统的实用性至关重要。本研究介绍了一种针对[具体名称未给出]的新方案，该方案结合了荧光原位杂交（FISH）和2'-脱氧-2'-氟-5-乙炔基尿苷（F-EdU）摄取及染色。我们的方案允许对基因表达进行空间检测并同时识别增殖细胞。F-EdU与干细胞标记物的双重标记揭示了在不同解剖区域，特别是头部和颈部的独特表达模式。我们展示了无需通透处理的最佳标记效果，简化了方案。我们还通过对其他组织标记物使用FISH证明了其通用性。该方案被应用于进行大量谱系追踪，揭示干细胞可在3天内分化为神经元和皮层细胞。我们的方案为研究[具体名称未给出]中的基因表达和细胞增殖提供了一种重要工具，为寄生绦虫生物学提供了有价值的见解，并可能为人类寄生绦虫的研究开辟新途径。
