The mechanical properties of the brain change dynamically throughout development, aging, and disease progression, influencing cellular behavior and tissue function. Glial cells, including astrocytes, microglia, and oligodendrocytes, experience distinct mechanical environments within the brain and respond to these cues in diverse ways. Understanding how mechanical factors influence glial cell behavior and the underlying mechanotransduction mechanisms provide valuable insights into glial function in both physiological and pathological contexts. In this chapter, we discuss how mechanical cues, especially substrate stiffness, regulate the behavior of astrocytes, microglia, and oligodendrocytes, affecting their activities and functions. We also summarize the mechanotransduction signaling pathways involved in these processes, including mechanosensitive ion channels such as Piezo1 and transient receptor potential (TRP) channels, as well as other mechanisms involving integrins, Rho GTPases, cytoskeletal dynamics, and YAP/TAZ. By integrating these findings, we provide a comprehensive overview of how glial cells sense and respond to mechanical stimuli, highlighting the broader implications for neuroscience and the development of therapeutic strategies targeting neurological diseases.