RESUMO
Salivary glands are essential for the maintenance of oral health by providing lubrication and antimicrobial protection to the mucosal and tooth surfaces. Saliva is modified and delivered to the oral cavity by a complex multifunctional ductal system. During development, these ducts form as solid tubes, which undergo cavitation to create lumens. Apoptosis has been suggested to play a role in this cavitation process along with changes in cell polarity. Here, we show that apoptosis occurs from the very earliest stages of mouse salivary gland development, much earlier than previously reported. Apoptotic cells were observed in the center of the first epithelial stalk at early-stage embryonic day 12.5 (E12.5) according to both TUNEL staining and cleaved caspase 3 immunofluorescence. The presumptive lumen space was highlighted by the colocalization of a predictive lumen marker, cytokeratin 7. At E14.5, as lumens start to form throughout the glands, apoptotic expression decreased while cytokeratin 7 remained positive. In vitro inhibition of all caspases in E12.5 and E13.5 salivary glands resulted in wider ducts, as compared with the controls, and a defect in lumen formation. In contrast, no such defect in lumen formation was observed at E14.5. Our data indicate that apoptosis is involved during early stages of gland formation (E12.5 onward) and appears important for shaping the forming ducts.
Assuntos
Apoptose/fisiologia , Morfogênese/fisiologia , Ductos Salivares/embriologia , Clorometilcetonas de Aminoácidos/farmacologia , Animais , Caspase 3/análise , Caspase 3/efeitos dos fármacos , Inibidores de Caspase/farmacologia , Polaridade Celular/fisiologia , Desenvolvimento Embrionário/fisiologia , Epitélio/embriologia , Marcação In Situ das Extremidades Cortadas , Queratina-7/análise , Camundongos , Técnicas de Cultura de Órgãos , Ductos Salivares/efeitos dos fármacos , Glândula Submandibular/embriologiaRESUMO
Immunoexpression of the extracellular matrix (ECM) proteins laminin, fibronectin, tenascin and types I, III and IV collagen was analyzed in the major and minor salivary glands of seven human fetuses at different gestational ages. The results showed the presence and localization of laminin, collagen IV and fibronectin around glandular structures at all stages of development. Tenascin was only detectable around excretory ducts. In the earliest stages of development, type I and type III collagen were presented as fine fibers delineating the glandular structures and delimiting the extension of the future lobule. As glandular development proceeded, the lobule was gradually filled with collagens and glandular tissue.
Assuntos
Proteínas da Matriz Extracelular/análise , Glândulas Salivares/embriologia , Anticorpos , Colágeno Tipo I/análise , Colágeno Tipo III/análise , Colágeno Tipo IV/análise , Feto , Fibronectinas/análise , Idade Gestacional , Humanos , Imuno-Histoquímica , Laminina/análise , Ductos Salivares/embriologia , Glândulas Salivares Menores/embriologia , Tenascina/análiseRESUMO
The presence of an epithelium at different stages of proliferation and differentiation raises interesting questions concerning the histogenesis, cell turnover and differentiation of normal salivary glands. In order to expand knowledge of these aspects, we investigated the expression of cytokeratins (CKs) 7,8,10,13,14,16,18 and 19, vimentin (VIM), and smooth muscle actin (SMA) in developing human minor salivary glands using monoclonal antibodies. Labial, buccal, palatine, and lingual salivary glands and those from the floor of the mouth were obtained from human fetuses (forensic postmortem) ranging in age from gestational weeks 10 to 29. Serial sections, 3 microm thick, were immunostained using a strepto-avidin-biotin technique. Reactivity for all antibodies was negative in the salivary gland epithelium during the developmental stages of bud formation, cord growth, and branching of cord. During canalization and cytodifferentiation, the glandular epithelial cells showed a positive reaction to some CKs and SMA. Cytokeratins 7, 8, 18, and 19 showed strong labeling in luminal duct cells that exhibited some degree of morphological differentiation. Myoepithelial cellc were recognized by antibodies to SMA. Cytoskeletal protein expression changes according to the cell type, degree of differentiation, and stage of morphological development of the glandular structure. These changes occur independently of the localization of the gland.