<p>Elevated glucose metabolism triggers two primary processes that lead to β-cell depolarization and insulin secretion: the closure of ATP-sensitive K⁺ channels via ATP-dependent mechanisms and the activation of mechanosensitive channels (MSCs) due to cell swelling. However, the identity of these MSCs remains unclear. In this study, we found that TMEM63B is a stretch-activated cation channel (SAC) crucial for regulating insulin secretion in response to elevated glucose levels. TMEM63B is abundantly expressed in β-cells, and its deletion impairs insulin secretion triggered by high glucose. High glucose levels typically increase Ca²⁺ influx and firing frequency in β-cells, a response largely eliminated when TMEM63B is deleted. Mechanistically, glucose metabolism induces cell swelling and activates TMEM63B, which, in turn, leads to β-cell depolarization and insulin secretion. In conclusion, our findings demonstrate that TMEM63B is an SAC essential for regulating insulin secretion in response to elevated glucose levels.</p>