Mechanism of acid clearing in ferret esophageal submucosal gland under gastroesophageal physiological homeostasis
LI Bo1, HUANG He2, WANG Lei1, ZHOU Haining1
1. Department of Gastrointestinal Surgery, General Hospital of Ningxia Medical University, Yinchuan 750000, China; 2. Department of Gastrointestinal Surgery, the First Affiliated Hospital of Shanxi Medical University, Taiyuan 030000, China
Abstract：Objective To investigate the mechanism of acid clearing in ferret esophageal submucosal gland under gastroesophageal physiological homeostasis.Methods The esophagus of adult ferrets aged 6-8 months and juvenile ferrets aged 1, 3, 5, 1 and 2 weeks after birth were selected and made into frozen and paraffin tissues in the Department of Pathology of General Hospital of Ningxia Medical University. Hematoxylin and eosin staining (HE staining) and periodized SCHFF staining (PAS) were used to analyze the morphology of esophageal submucosal glands (SMG) in adult ferrets. SMG binding of ferret esophagus was detected by LECTIN ; the expressions of MUC5B and MU5AC were detected by immunohistochemistry and immunofluorescence. The expression of cystic fibrosis transmembrane conductance regulator (CFTR) in adult ferret esophageal SMG was detected by tissue immunofluorescence in situ hybridization. Immunofluorescence was used to detect the expression of lymphocyte binding enhancement factor (LEF1) in SMG of neonatal ferrets on day 1, 3, 5, 7, and 14, and to analyze the expression of LEF1 in esophageal epithelial cells of juvenile ferrets at different developmental stages.Results HE staining showed that the SMG of the submucosal glands of the adult ferret esophagus was the connective tissue beneath the mucosal muscle layer of the esophagus, and PAS staining showed that the SMG had a secretory function; different cell types in the SMG of the adult ferret esophagus specifically bound different lectins, and the mucosal cells specifically bound Dolichos biflorus (DBA), ECL, GSL Ⅰ, GSL Ⅱ, SOYBEAN, tacalin biotylated, ULEX, Triticum vulgare Wheat germ agglutinin (WGA); specific binding of lectins GSL I and GSL Ⅱ in ductal cell membranes and positive expression of lectins ECL, PNA and WGA in epithelial cell membranes; lectins Mucin staining showed positive staining for MUC5B MU5AC in the cytoplasm of adult ferret esophageal glandular mucus cells and in the lumen of the gland. in situ hybridisation showed that CFTR was positively expressed in the SMG of the adult ferret esophagus; as the gland developed, LEF1 was expressed in the developing gland from 3 days to 1 week after birth, but disappeared when the gland differentiated and matured.Conclusions In ferret SMG, CFTR expression can regulate HCO3- secretion in steady state, and SMG can secrete mucus containing mucins MUC5B and MU5AC, which is helpful for esophageal acid clearance and mucosal protection. The changes of LEF1 expression in SMG under developmental conditions suggest that SMG may regulate the maintenance of homeostasis through WNT signaling LEF1.
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