Cannabinoid receptors (CBRs) belong to the G protein-coupled receptor superfamily and are widely expressed within the body. Although the expression of CBRs was predicted in nonexitable acinar cells, the molecular identification of different receptor subtypes in acinar cells has not been performed and the physiological consequences of their activation in salivary glands remain to be fully established. To elucidate these issues in the acinar cells of submandibular salivary glands, we used different molecular biological and fluorescent imaging techniques. Experiments were performed on acinar cells freshly isolated from the submandibular salivary glands of deeply isoflurane-anesthetized Wistar rats deeply anesthetized with isoflurane (2 %). The animals were used in accordance with protocols that were approved by the Animal Care and Use Committee at Bogomoletz Institute of Physiology, Ivan Franko National University, and The University of Dundee. By utilizing a set of molecular biology approaches we have demonstrated the prominent expression of classical CB1Rs and CB2Rs in acinar cells of the rat submandibular gland. In addition, we also identified expression of the putative novel cannabinoid receptor GPR55 in these cells. Using Ca2+ imaging technique, we found that the activation of different CBRs subtypes triggers [Ca2+]cyt signalling in acinar cells by distinct pathways, involving Ca2+ release from the endoplasmic reticulum (ER) and store-operated Ca2+ entry. Altogether, our findings directly demonstrate a functional expression of different CBRs subtypes, especially CB1Rs, CB2Rs, and GPR55, in acinar cells from the submandibular salivary glands. The presence of CBR subtypes in acinar cells suggests that they may play a novel role in regulating the activity of the salivary gland.