This review focuses on the cellular mechanisms underlying the secretion of incretin hormones, particularly glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP), which play crucial roles in glucose homeostasis and appetite regulation. Enteroendocrine cells (EECs) located along the gastrointestinal (GI) tract secrete these hormones in response to nutrient stimuli. The review discusses the various signaling pathways involved in hormone secretion, including ion channels, nutrient transporters, and G-protein-coupled receptors (GPCRs). It highlights the complexity of EECs, where multiple hormones can be secreted from a single cell type, and the regional distribution of hormones along the GI tract. The review also explores the use of different models, such as cell lines, primary epithelial cultures, and organoids, to study EEC function. Key signaling pathways, including calcium channels, cyclic adenosine monophosphate (cAMP), and GPCR activation, are discussed in detail. The review further examines the postprandial sensing mechanisms of EECs, focusing on GPCRs and transporters that respond to nutrients like carbohydrates, fats, and proteins. It concludes by discussing the potential of targeting EECs for the treatment of type 2 diabetes and obesity, emphasizing the need for further research to understand the complex interactions within EECs and to develop effective therapeutic strategies.This review focuses on the cellular mechanisms underlying the secretion of incretin hormones, particularly glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP), which play crucial roles in glucose homeostasis and appetite regulation. Enteroendocrine cells (EECs) located along the gastrointestinal (GI) tract secrete these hormones in response to nutrient stimuli. The review discusses the various signaling pathways involved in hormone secretion, including ion channels, nutrient transporters, and G-protein-coupled receptors (GPCRs). It highlights the complexity of EECs, where multiple hormones can be secreted from a single cell type, and the regional distribution of hormones along the GI tract. The review also explores the use of different models, such as cell lines, primary epithelial cultures, and organoids, to study EEC function. Key signaling pathways, including calcium channels, cyclic adenosine monophosphate (cAMP), and GPCR activation, are discussed in detail. The review further examines the postprandial sensing mechanisms of EECs, focusing on GPCRs and transporters that respond to nutrients like carbohydrates, fats, and proteins. It concludes by discussing the potential of targeting EECs for the treatment of type 2 diabetes and obesity, emphasizing the need for further research to understand the complex interactions within EECs and to develop effective therapeutic strategies.