Beta cell secreted GABA sets appropriate insulin secretion by modulating islet calcium oscillations

Gamma-aminobutyric acid (GABA) is produced in pancreatic beta cells and is implicated in modulating islet function, yet its precise physiological role remains uncertain. This study aimed to determine the function of endogenous beta cell-derived GABA on insulin secretion and islet calcium dynamics by developing a conditional beta cell-specific knockout of GABA-synthesizing enzymes (GAD65 and GAD67). Conditional knockout mice ( Gad βKO ) lacking both GAD65 and GAD67 specifically in pancreatic beta cells were generated. Glucose-stimulated insulin secretion was measured in isolated islets and in vivo and islet Ca 2+ oscillations were recorded using calcium imaging. The effects of GABA and its receptor agonists/antagonists were tested under various glucose conditions. Additional analyses were performed in high-fat diet-fed mice and human islets from donors with and without type 2 diabetes (T2D). Gad βKO islets were devoid of GABA and showed excessive insulin secretion in response to glucose without anatomical changes in islet composition. These islets had defective Ca 2+ oscillations, with prolonged active phases and reduced amplitudes. GABA application suppressed Ca 2+ oscillations, an effect mediated by GABA A and GABA B receptors. High-fat diet-fed and T2D human islets were unresponsive to GABA and exhibited impaired Ca 2+ oscillations. This is the first study using a beta cell-specific GAD65/GAD67 knockout model. Endogenous beta cell-derived GABA is critical for modulating insulin secretion by maintaining proper Ca 2+ oscillation dynamics. GABA signaling likely operates as a delayed negative feedback mechanism that reinforces oscillatory homeostasis in islets. The loss of GABA responsiveness, as seen in metabolic stress or T2D, may contribute to islet dysfunction. This work establishes GABA as a key regulator of islet rhythm and glucose responsiveness. • First functional analysis of islets lacking endogenous GABA • Establishment of a conditional, beta cell-specific GAD knockout model • Clarification that GABA is dispensable for beta cell mass, but essential for dynamic function • Identification of GABA as a modulator of islet calcium oscillations