The expression of the gap-junction proteins connexin 32 and 43 in the developing rat submandibular gland was determined using reverse transcription-polymerase chain reaction and immunohistochemistry. Sprague-Dawley rats from the 17th gestational day through to the 28th postnatal day were used. Connexin 43 gene expression was detected on the 17th gestational day. Connexin 32 gene expression was also detected on the 17th gestational day but less intensely. Immunolabelling for connexin 43 was also found in the developing submandibular gland from the 17th gestational day. Prenatally the most immunoreactive areas for connexin 43 were located in the periphery of terminal tubules, but some staining was discernible between the cells in terminal buds. In the postnatal period, reactive areas were located at both the periphery of acinus-like structures and around the intercalated duct. This is consistent with the known association of connexin 43 with myoepithelial cells. Connexin 32 immunostaining was first detected in the developing submandibular glands on the 18th day of prenatal development. Positive staining was present on the lateral side of the proacinar and mature acinar cells. The number of immunoreactive areas per cell increased during early development followed by a significant decrease perinatally. During postnatal development the density of areas again showed a pattern of increase. These results suggest that connexin 43 is associated with growth and differentiation in the pre- and perinatal periods and also with the contractile function of myoepithelial cells in the postnatal period of the developing submandibular gland. It is also implied that the number of connexin 32-positive areas may correspond to an increase or decrease in the number of proacinar and mature acinar cells.