Department of Food Science and Nutrition, Faculty of Food Engineering, University of Campinas, Campinas, Brazil.
Division of Biology and Biological Engineering, California Institute of Technology, 1200 E. California Boulevard, Pasadena, CA, USA.
STAR Protoc. 2022 Jun 18;3(3):101486. doi: 10.1016/j.xpro.2022.101486. eCollection 2022 Sep 16.
Gut microbiota influences neurodevelopment, behavior and contributes to neurodegenerative disorders. One possible mechanism is the direct modulation of calcium (Ca) signaling and protein homeostasis in enteroendocrine cells (EECs), a component of the gut epithelium. Here, we present a protocol to isolate fractions of conditioned media (CM) from the anaerobic bacteria and the utilization of this CM to monitor Ca fluctuation in EECs by imaging. This protocol can be adapted and applied to various bacterial cultures and cell types. For complete details on the use and execution of this protocol, please refer to Amorim Neto et al. (2022).
肠道微生物群影响神经发育、行为，并导致神经退行性疾病。一种可能的机制是直接调节肠内分泌细胞 (EECs) 中的钙 (Ca) 信号和蛋白质动态平衡，EECs 是肠道上皮的一个组成部分。在此，我们提供了一个从厌氧细菌中分离条件培养基 (CM) 级分的方案，并利用该 CM 通过成像来监测 EECs 中的 Ca 波动。该方案可以适应并应用于各种细菌培养物和细胞类型。有关此方案的使用和执行的完整详细信息，请参阅 Amorim Neto 等人。(2022)。
