Measurement of rat gastric mucus secretion using ex-vivo gastric chamber system

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For the assessment of gastric mucus secreted from rat gastric mucosa into gastric lumen, we examined an ex-vivo gastric chamber system which had been developed by Takeuchi et al. To apply this system for the measurement of mucin secreted into the effluent solution, we had to develop a highly sensitive method to determine gastric mucin content. Finally we adopted an HPLC method to determine the content of hexosamine which constitutes the major carbohydarate component of mucin molecule. By using this method coupled with the removal treatment of low molecular weight hexosamine-containing materials including non-mucinous glycoproteins, it could be achieved to measure the time course changes of mucin secreted into the effluent solution. This method was applied to estimate the effect of carbachol administration on the mucus secretion in the rat gastric mucosa, and obtained a result that this drug to be an accelerator of mucus secretion into the gastric lumen.In the course of this study, we tried to obtain some monoclonal antibodies(MAbs) which are specific for the gastric mucin, and utilize them to the immunohistochemical method to determine the mucin content. We established several MAbs which are specific for the carbohydrate moieties of mucins present in the specific region of rat gastric mucosa. At present, we are trying to establish a method to assess the content of mucin originated from a specific site of gastric mucosa such as surface mucous cells, corpus gland mucous cells, antral gland mucous cells.