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Ultrastructure of Endocrine-Like Cells in Lamina Propria of Human Gastric Mucosa

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Ultrastructure of Endocrine-Like Cells in Lamina Propria of Human Gastric Mucosa Gut: first published as 10.1136/gut.22.7.534 on 1 July 1981. Downloaded from Gut, 1981, 22, 534-541 Ultrastructure of endocrine-like cells in lamina propria of human gastric mucosa* J STACHURA, W J KRAUSE, AND K J IVEYt From the Veterans Administration Medical Center, Long Beach, California; University of California Irvine, Irvine, California; Department of Anatomy, University of Missouri School of Medicine, Columbia, Missouri; and the Institute ofPathology, Copernicus Medical Academy, Krakow, Polantd SUMMARY Endocrine cells of gastric and gut mucosa are commonly thought to be present only within mucosal glands. In a previous report, we described argyrophilic cells in the lamina propria in 40 % of surgical gastric specimens, using light microscopy. All these patients had chronic gastritis. Argyrophilia, however, is a non-specific reaction which could occur in other than endocrine cells. The present study was undertaken to describe the ultrastructure of argyrophil cells in the lamina propria. In five patients with chronic gastritis, endoscopic biopsies were taken from the fundic, intermediate, and pyloric areas of the stomach. Single and/or clustered argyophil cells were seen by light microscopy in the lamina propria of the intermediate and pyloric are.as. On electron- microscopy, these cells had the following characteristics of endocrine-like cells: they were characterised by numerous electron dense granules in the cytoplasm, 100-300 nm in diameter; the cytoplasm contained poorly-developed rough endoplasmic reticulum and well-developed smooth endoplasmic reticulum with occasional vesicles. Immunostaining gave negative results for various gastrointestinal hormones. These ultrastructural characteristics of lamina propria cells are similar to endocrine cells of the APUD series. We conclude that endocrine-like cells occur in the lamina http://gut.bmj.com/ propria of the human stomach in the presence of chronic gastritis. Endocrine cells of gastrointestinal mucosa are Methods usually found within mucosal glands.' Light microscopic examination has suggested the Biopsies of five subjects (four female, one male, presence of endocrine cells in the lamina aged 25-47 years) were investigated by light and propria on October 2, 2021 by guest. Protected copyright. of intermediate and pyloric mucosa of patho- electron microscopy. Biopsies were chosen from logical human gastric mucosa.2 Argyrophilic 60 patients endoscoped during a three-month cells, singly or in groups, were found in the lamina period for investigation of gastric or duodenal propria of 22 of 56 surgical gastric specimens of ulcer disease or dyspepsia. A duplicate biopsy was patients operated on for gastric or duodenal taken from pyloric (about 2 cm from pylorus), ulcer or gastric cancer. All subjects with argyro- intermediate (7-9 cm from pylorus along greater philic cells in the lamina propria had chronic curva,ture), and body (mid-greater curvature) gastritis.2 The argyrophilic reaction, however, is areas in each patient. One biopsy was routinely non-specific, and thus it could only be hypothe- examined histologically, the other was collected in sised that these argyrophilic cells were endocrine Epon blocks. Biopsies from five of these 60 cells.2 This study presents ultrastructural subjects showing argyrophil positive lamina evidence for endocrine-like morphological char- proprial cells in the pyloric and/or intermediate acteristics of these argyrophilic cells in the area were chosen for ultrastructural investiga- gastric lamina propria of man. tions. *This work was supported in part by the CRC NIH Grant No. Tissue for histological examination was fixed in RR 00287-12 and Biomedical Grant No. RR 07053, and by the 10% neutral formalin and embedded in paraffin. Medical Research Service of the Veterans Administration. Paraffin sections were stained with haematoxylin tAddress for all correspondence and reprint requests: Dr K J Ivey, Gastroenterology Section (IIIG), Veterans Administration Medical and eosin (H and E). Periodic acid-Schiff (PAS) Center, 5901 East Seventh Street, Long Beach, California 90822, USA. reaction, with Grimelius method for argyrophilia,3 Received for publication 9 January 1981 and the Masson-Fontana4 argentaffin reaction 534 Gut: first published as 10.1136/gut.22.7.534 on 1 July 1981. Downloaded from Ultrastructure ofendocrine-like cells in lamina propria ofhuman gastric mucosa 535 Table Immunohistochemical investigations of endocrine-like cells in lamina propria by PAP methods5 Antiserum to Dilution Source used Gastrin I and II 1:500 Calbiochem Glucagon 1:1000 Calbiochem Somatostatin 1:300 A Arimura. Tulane Univ. School of Med, New Orleans, Louisiana, USA VIP 1:2500 S 1 Said, Depts of Internal Med. and Pharmacol., Univ. Texas, Dallas, USA Secretin 1:500 Calbiochem Pancreatic polypeptide 1:500 Lilly Res Labs, Indianapolis, (bovine) Indiana, USA Substance P 1:500 R H Ho, Dept of Anatomy, Ohio State Univ, Columbus, USA were performed. Tissue for ultrastructural examination was fixed in 2% glutaraldehyde in phosphate buffer, pH 7.4 at 4°C for two hours. Tissues were then washed in phosphate buffer, pH 74, and post- 1P... W fixed in 2% osmium tetroxide for two hours, dehydrated in a graded series of ethyl alcohol, Fig. 1 Groups ofargyrophil cells (large arrow) and embedded in Epon 812 resin. thin localised in the lamina propria. Single argyrophil cells Silver-grey also are observed within glandular epithelium (small sections were stained with uranyl acetate and arrow). x 260. lead citrate and examined on a Philips 300 elec- tron microscope operated at 80 KV. Immunostaining for gut hormones was per- like cells (Figs 3 and 4). They were characterised http://gut.bmj.com/ formed according to Sternberger's PAP method5 by round and oval electron dense granules 100- using formalin-fixed paraffin sections. Antisera 200 nm in diameter in some cells (type I, Fig. 5) used, dilution and source are shown in the Table. and 150-300 nm in other cells (type II, Fig. 6). The nuclei of these cells were oval and slightly Results irregular in shape, with well-developed, smooth endoplasmic reticulum, with occasional vacuole LIGHT MICROSCOPY formation. Mitochondria were not very numer- on October 2, 2021 by guest. Protected copyright. Chronic gastritis was present in the biopsies of ous. Lysosomal bodies containing electron the pyloric and intermediate areas in all five dense material were also noticed. Cell boundaries subjects showing argyrophil positive lamina pro- usually were poorly defined, as they were inter- prial cells. Focal intestinal metaplasia was pre- mingled with collagen fibres. Occasionally, the sent in two of these subjects. Focal superfici'al endocrine-like cells formed small clusters of two gastritis occurred in the fundic area. to five cells (Figs 7 and 8). Some cells were Based on the Grimelius argyrophil reaction, found close to the basal lamina of the gastric we found numerous positive staining endocrine glands; others, however, were situated deep in cells in the gastric pyloric and fundic glands as the connective tissue of the lamina propria, far has been well described by others. In addition, away from the glandular cells. We observed single and/or groups of argyrophil cells were nerve fibres close to some endocrine-like cells. found in the lamina propria as well, in the Other endocrine-like cells were situated adja- pyloric and intermediate zones (Fig. 1). The posi- cent to small blood vessels (Fig. 8). tion of these cells in the lamina propria of the A moderate inflammatory infiltrate with mucosa is shown in Fig. 2. The Masson-Fontana numerous plasma cells was observed within the argentaffin reaction was negative. lamina propria, as were granulated mast cells. Parietal and mucous cells present in the gastric ELECTRONMICROSCOPY glands were morphologically normal. Endocrine Ultrastructural examination showed these argyro- cells in the normal distribution within gastric phil cells in the lamina propria to be endocrine- glands were primarily enterochromaffin-like Gut: first published as 10.1136/gut.22.7.534 on 1 July 1981. Downloaded from 536 Stachura, Krause, and Ivey *wk http://gut.bmj.com/ s \ff*tThas w \E Fig. 2 Histological picture of the pyloric mucosa with bracketed area occupied by lamina proprial argyrophil cells. Hand E, x 100. cells, enterochromaffin cells, and D, cells (classi- may be found in the lamina propria."-10 on October 2, 2021 by guest. Protected copyright. fied according to Lausanne 1977 classification'). Ultrastructural investigations of rat gastric The endocrine-like cells found in the lamina lamina propria have demonstrated directly propria did not resemble any of the endocrine innervated, granulated, endocrine-like cells.6-8 10 cells normally found in the stomach. Some of the Granulated cells in the duodenal lamina propria cells studied (type I) were similar to D, cells by of the human fetus were also reported.9 In these the size of granules. Immunostaining for VIP was studies, granulated cells were closely connected negative, however, in our formalin-fixed, with unmyelinated nerve fibres. The authors paraffin-embedded material. postulated that these 'neuroendocrine' cells may have a possible neuroendocrine function. To our IMMUNOSTAINI NG knowledge, there has been no published ultra- Immunohistochemical reactions for gastrin, structural study of endocrine-like cells in human glucagon, somatostatin, pancreatic polypeptide, adult gastric lamina propria. secretin, substance P, and VIP were negative. Our present study clearly
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