LOCALIZATION AND IDENTIFICATION OF ENDOCRIN CELLS IN THE ALIMENTARY CANAL OF LATEOLABRAX JAPONICUS

Localization of immunohistochemistry action of hormone in the alimentary canal of Dsteichehyes has been reported by many scientists.11 kinds of odd endocrine cells were differentiated in digestive system from many stomachless teleast and stomached teleost fish.The studies showed that distribution of immunity endocrine cells in fish alimentary canal was very complexities.In this study, by using immunohistochemistry Elivison method as well as 5 Kinds of anti-mammal serum such as 5-hydroxytryptamine(5-HT), somatostatin(Som),gastrin(Gas), glucagon(Glu) antisera, pancreatic polypetide(PP), the endocrin cells in the alimentary canal of L. japonicus were studied with immunohistochemical localization technique.Four L.japonicus were collected from the supermarket Fuzhou in weighting about 600g, which was vivisection from abdomenand sampled from different parts of alimentary canal including the pylorus stomach, apical and middle parts of the gut folds,esophagus, cardiac stomach, gastric pouch and hind intestine,the apical and middle parts of the gut folds.The samples were fixed by neutral formalin for 24 hours, then general dehydrated, paraffin wax embed, 6um section and sticking on glass platen, 40℃ drying for 24h.Coloration step of immunohistochemistry: general slicing and dehydration with paraffin wax, watering 3 times by means of PBS. 1): 50ul H2O2was given and incubation 10 min under roomtemperature; 2):watering 3 times by PBS. 3):50μl no immunity animal blood serum was given and incubation 10 min under room temperature; no washing because of unspecific immunity background; 4): each section was given 50μl first antibody under 4 ℃ and staying overnight; 5):watering 3 times by means of PBS.6): 5ul of second antibody was given in each section for 10-30min under room temperature; 7):watering 3 times by means of PBS. 8):DAB coloration for 5-10min under room temperature; 9):washing by tap water, coloration again by Hematoxylin, dehydration and enveloping by neutral balata. In this experiment, continuing slice up and take picture on correspond position, first antibody serumwas replaced by PBS,other steps synchronizing like above.5-hydroxytryptamine (5-HT) cell: The result showed that 5-HT cells are distributed in the esophagus, cardiac stomach, gastric pouch, pylorus stomach,which were not found in every part of intestinal canal.We can see 5-HT cells are distributed in simple columnar epithelial cell of esophagus mucous membrane, which looked like triangle and pyramid long shuttle shape, dissociated end run out to esophagus and 5-HT cells arenot distributed in the areaof flat epithelium cells. 5-HT cells of stomach distributed between mucous membrane and glandular epithelium epithelial cell also looked like pyramid long shuttle and triangle shape,and cell project run out to epithelium epithelial cell of stomach cavity and acinus cavity, some cells looked like oblong shape but no cell project can be seen.SST cell:The experiment revealed that many SST cells are distributed in cardiac stomach and pylorus stomach, a few in the esophagus, and the cells could not found in the intestine We can see SST cells are distributed in simple columnar epithelial cell of esophagus mucous membrane, which looked like triangle and pyramid long shuttle shape,dissociated end run out to esophagus and SST cells are not distributed in the area of esophagus mucous membrane flat epithelium cells.SST cells of stomach only few ness located between membrane epithelial cells, the shape of cells looked like taper, dissociated end run out to esophagus.Most of SST cells distributed in the shallow of gastric gland epithelial cell, some cells looked like ellipse shape, dissociated end run out to acinus cavity.The other dissociated end run out to the basement membrane of acinus cavity, wheresome cells located and adjacent to gland epithelial cell,the cells looked like round and triangle shape.GaS cell: In this study we also can see many Gas cells distributed in pylorus stomach, the apical and middle parts of the gut folds,which were not found in every part of the gut folds.GaS cell in pylorus stomach distributed between stomach membrane epithelial cells.The cells looked like stick and anomaly shape, dissociated end run out to acimus cavity and cling to acinus membranesometime. GaS cell between columnar epithelial cell of intestine looked like long stick and egg shape, and a few dissociated end run out to intestine cavity.Glu cell:The result also showed that Glu cell only distributed in the apical and middle parts of the gut folds,which were not found in every part of the gut folds. Glu cells located between columnar epithelial cell and cupular cell of intestine, the cell looked like ellipse shape, and dissociated end run out to intestine cavity.