Meena Netra Pal, Kimmel Alan R
Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, MD, 20892, USA.
Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Building 50, Room 3351, 50 South Drive, Bethesda, MD, 20892, USA.
Methods Mol Biol. 2016;1407:141-51. doi: 10.1007/978-1-4939-3480-5_11.
Dictyostelium discoideum has proven an excellent model for the study of eukaryotic chemotaxis. During growth in its native environment, Dictyostelium phagocytose bacteria and fungi for primary nutrient capture. Growing Dictyostelium can detect these nutrient sources through chemotaxis toward the metabolic by-product folate. Although Dictyostelium grow as individual cells, nutrient depletion induces a multicellular development program and a separate chemotactic response pathway. During development, Dictyostelium synthesize and secrete cAMP, which serves as a chemoattractant to mobilize and coordinate cells for multicellular formation and development. Separate classes of GPCRs and Gα proteins mediate chemotactic signaling to the chemically distinct ligands. We discuss common and separate component responses of Dictyostelium to folate and cAMP during growth and development, and the advantages and disadvantages for each. As examples, we present biochemical assays to characterize the chemoattractant-induced kinase activations of mTORC2 and the ERKs.
盘基网柄菌已被证明是研究真核生物趋化性的优秀模型。在其原生环境中生长时，盘基网柄菌通过吞噬细菌和真菌来获取主要营养物质。生长中的盘基网柄菌可通过向代谢副产物叶酸进行趋化作用来检测这些营养源。尽管盘基网柄菌以单个细胞形式生长，但营养物质耗尽会诱导多细胞发育程序和一条独立的趋化反应途径。在发育过程中，盘基网柄菌合成并分泌cAMP，cAMP作为一种化学引诱剂来动员和协调细胞进行多细胞形成与发育。不同类别的GPCR和Gα蛋白介导对化学性质不同的配体的趋化信号传导。我们讨论了盘基网柄菌在生长和发育过程中对叶酸和cAMP的共同及不同组分反应，以及各自的优缺点。作为示例，我们展示了用于表征化学引诱剂诱导的mTORC２和ERK激酶激活的生化分析方法。
