Author: 〇李 傲男1、高 靖1、藤井 慎介2、清島 保2、自見 英治郎1,3
Affiliation: 1九大 院歯 口腔細胞工学、2九大 院歯 口腔病理学、3九大 院歯 OBTセンター
Abstract:
The salivary glands of rodents are not fully developed at birth, and cell differentiation involved in its function occurs after birth. p130Cas (Crk-associated substrate protein) is an adapter protein that forms a complex with various proteins in cells by growth factors and integrin signals. We have previously reported that p130Cas is not involved in osteoclast formation, but is essential for the formation of ruffled border involved in the secretion of acids and proteolytic enzyme. Since we found that p130Cas was expressed in ductal epithelial cells including the granular duct of the submandibular gland by immunohistochemical technique, we considered that it may play an important role in salivary secretion. In this study, we generated epithelial tissue-specific p130Cas-deficient (p130Cas cKO) mice using Keratin14-Cre mice and investigated the physiological role of p130Cas in salivary gland development. The submandibular glands of 5-week-old wild-type (WT) and p130Cas cKO mice were prepared, and their size and weight were measured. Mice were anesthetized and stimulated with pilocarpine and then saliva was collected to measure the saliva secretion and to analyze saliva components. For histological analysis, salivary glands were fixed and then stained with H&E or with antibodies against p130Cas, E-cadherin, and GM130. To label proliferating cells, EdU was intraperitoneally injected to 5-week-old WT and p130Cas cKO mice. After 8 hours, salivary glands were fixed and detected with click-iT plus kit.The size and weight of submandibular glands of p130Cas cKO mice were lower than those of WT mice. The saliva secretion and the amount of amylase in saliva were decreased in p130Cas cKO. Histological analysis showed immature development of the granular convoluted tubule of the submandibular glands in p130Cas cKO mice. Furthermore, it was revealed that the secretory granules contained in the granular convoluted ductal epithelial cells were significantly reduced in the p130Cas cKO mice. Deficiency of p130Cas caused the cytoplasm localization of E-cadherin which should be expressed on the cell membrane of granular convoluted ductal epithelial cells and disturbed the intracellular localization of the cis-Golgi matrix protein GM130. The incorporation of EdU into submandibular gland was decreased in p130Cas cKO mice compared to WT mice. Thus, p130Cas plays an important role in the normal development of the granular convoluted tubule of salivary gland and in the saliva secretion.
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