Study Of Calcitonin In Rat Submaxillary Gland And In Vitro Localization Cells

Objective:Submandibular gland with endocrine function can secrete a variety of biologically active substances,becase the thyroid and submandibular glands are from the same gastrointestinal pancreatic endoderm differentiation and formation,their structure and function may be closely related.Recent studies have reported that submandibular glands can secrete calcitonin (CT),which suggested submandibular gland secretion of calcitonin regulate calcium and phosphorus metabolism in the body.Through this study,neonatal rat submandibular gland cells is cultured with the natural method of in vitro , and immunohistochemistry staining cultured cells,situ hybridization confirmed the presence of calcitonin,using enzyme-linked immunosorbent assay Experimental determination of calcitonin content.Verify that rat submandibular gland cells secrete calcitonin, and further confirmed that the endocrine function of submandibular gland.Methods: (1) Were taken the submandibular gland of rat,making paraffin tissue sections for immunohistochemistry,confirmed the submandibular gland cells secrete calcitonin. (2) Establishing rat submandibular gland cells in primary and passaged in vitro culture system, using immunocytochemistry,anti-cytokeratin and anti-CT staining cultured cells,watch characteristics of cells and distribution of calcitonin in the cell. (3) Digoxigenin labeled DNA of calcitonin in situ hybridization method verify the submandibular gland cells secrete calcitonin and gene mapping. (4) Enzyme-linked immunosorbent assay (ELISA) determine calcitonin content.Results:(1) On rat submandibular gland tissue slices observed:Rat submandibular gland granular convoluted tubule (GCT),intercalated duct,striated duct andinterlobular duct epithelial cells were respectively CT positive in the immune response,the reaction product distribute in the cytoplasm.(2) The cultured cells were flat growth triangle,square,round,rope-shaped and so on,like paving stone-like paste on the sidewall.cytokeratin is dyed and markers of epithelial cells. (3)From staining of cells and digoxigenin labeled DNA probe,observed the calcitonin secretion is in the cytoplasm, the nucleus-free expression. (4) On the basis of the above experiment using enzyme-linked immunosorbent assay (ELISA) measured levels of calcitonin.Conclusions: (1) From tissue slices of rat submandibular gland,clear secretion of calcitonin and their morphological structure and composition.(2) The establishment of a rat submandibular gland cells in primary and passaged in vitro culture system, immunocytochemistry and in situ hybridization confirmed that calcitonin secretion in submandibular gland cells. (3) ELISA test determined the content of submandibular gland secretion of calcitonin.