HPC-1/syntaxin1A (STX1A) is considered to regulate exocytosis in neurones and endocrine cells. Previously, we reported that STX1A null mutant (STX1A KO) mice unexpectedly showed normal glutamatergic and GABAergic fast synaptic transmission but exhibited disturbances in monoaminergic transmission, such as serotonin, 5-hydroxytryptamine (5-HT), which may induce attenuation of latent inhibition. These results suggest that STX1A may contribute to dense-core vesicle exocytosis in vivo. Thus, we hypothesised that the lack of STX1A might affect the secretion of several hormones, as also mediated by dense-core vesicles exocytosis. In the present study, we focused on the hypothalamic-pituitary-adrenal (HPA) axis, which is a neuroendocrine system that regulates responses to stress stimuli and is considered to be associated with neuropsychiatric disorders. Specifically, we examined whether the HPA axis is impaired in STX1A KO mice. Interestingly, plasma concentrations of both corticosterone (CORT) and adrenocorticotrophin hormone (ACTH) during the resting condition decreased in STX1A KO mice compared to WT mice. Additionally, elevated plasma CORT, ACTH and corticotrophin-releasing hormone (CRH) which were usually observed after acute restraint stress, were also reduced in STX1A KO mice. We also observed the suppression of 5-HT-induced CRH release in STX1A KO mice in vitro. Furthermore, an in vivo microdialysis study revealed that the elevation of extracellular 5-HT in the hypothalamus, which was induced by the selective serotonin reuptake inhibitor, fluoxetine, was significantly reduced in STX1A KO mice compared to WT mice. 5-HT elevation in the hypothalamus, which was induced by acute restraint stress, was also reduced in STX1A KO mice. Finally, STX1A KO mice showed abnormal behavioural responses after mild restraint stress. These results indicate that the lack of STX1A could induce dysfunction of the HPA axis, and the deficit may result in abnormal behavioural properties, such as unusual responses to stress stimuli.