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Aim: To investigate the ultrastructural localization of atrial natriuretic peptide(ANP)-synthesizing cells and the relationship between ANP-synthesizing cells and microvessels in rat gastric mucosa. Methods: Immunohistochemistry techniques and postembedding immunoelectron microscopy techniques were used to validate the findings regarding the expression of ANP-synthesizing cells and the ultrastructural localization of ANP-synthesizing cells in the gastric mucosa. Histochemistry techniques and the tannic acid-ferric chloride method (TA-Fe staining method) were used to reveal microvessel density and the distribution of ANPsynthesizing cells in different regions of the stomach. Results: Cells expressing ANP were localized and ANP-synthesizing cells were identified as enterochromaffin (EC) cells in the gastric mucosa. ANP-synthesizing cells existed in different regions of the stomach. The percentage ANP-synthesizing cells in the mucosa was greatest in the fundus (46.7%±5.3%), intermediate in the antrum (40.1%±4.5%), and least in the body (21.6%±3.6%). There was a positive relationship between the percentage of ANP-synthesizing cells and the density of microvessels in the antral mucosa, but not in the fundus or body mucosa. Conclusion: ANP is synthesized by EC cells in rat gastric mucosa, and ANP-synthesizing cells are most dense in the gastric fundus. ANP may act not only as a regional autocrine and/or paracrine regulator, but also as an endocrine regulatory peptide in the gastrointestinal tract.