Argentaffin Research Articles

Sites of prealbumin production in the human fetus using the indirect immunoperoxidase technique.

An indirect immunoperoxidase technique was used to investigate the site of prealbumin synthesis in the human fetus from between 8 and 43 weeks. Prealbumin staining was noted in the A cells of the fetal pancreas, both in the primary and secondary generations of the islets of Langerhans, from as early as 12 weeks gestation. Electron microscopy localized the staining to the cytoplasmic secretory granules. Prealbumin was also present in the gastrointestinal mucosa in cells which had a distribution similar to that of argentaffin cells. Positive staining was noted in the lining epithelium of the renal proximal convoluted tubules, but ultrastructural studies indicated that this was probably related to reabsorption, and not to synthesis.

Endocrine cells in extrahepatic bile duct carcinoma.

A total of 44 extrahepatic bile duct carcinomas comprising 13 well-differentiated adenocarcinomas, 25 moderately differentiated adenocarcinomas, and 6 poorly differentiated adenocarcinomas were examined histologically and immunohistochemically for somatostatin, gastrin, and glicentin. Argyrophil cells, argentaffin cells, and somatostatin- and gastrin-immunoreactive cells within the tumor were detected in 46.2%, 15.4%, 23.1%, and 15.4% of well-differentiated adenocarcinomas, and in 16.0%, 8.0%, 12.0%, and 4.0% of moderately differentiated adenocarcinomas, respectively. No tumor tissues of poorly differentiated adenocarcinomas contained endocrine cells. A statistically significant difference in the frequency of argyrophil cells was observed between well and poorly differentiated adenocarcinoma. The incidence of argyrophil cells and somatostatin-immunoreactive cells in nonneoplastic mucosa adjacent to well-differentiated adenocarcinoma was higher than in that adjacent to poorly differentiated adenocarcinoma. Glicentin-immunoreactive cells could not be demonstrated either in tumor tissue or in nonneoplastic mucosa of the extrahepatic bile duct. With reference to the histogenesis of extrahepatic bile duct carcinoma, it was assumed from these results that the development of well-differentiated adenocarcinoma might be closely related to the occurrence of endocrine cells and that poorly differentiated adenocarcinoma might develop from ordinary mucosa.

Human colonic substance P-producing cells are a separate population from the serotonin-producing enterochromaffin cells.

Human colonic mucosa was immunostained with antibodies against substance P to identify the endocrine cells containing this peptide in the mucosal glands. Dual immunohistochemical and histochemical studies were also carried out to determine whether these cells are enterochromaffin cells and contain serotonin as claimed in the literature. The results obtained indicate that the normal human colonic substance P-producing cells are not argentaffin cells, nor do they contain serotonin. In addition, they are also negative both for several silver and for other techniques commonly used to identify digestive endocrine cells. They are positive, however, with the argyrophilic technique of Churukian-Schenk. It is concluded, therefore, that the substance P-producing cells of the human colonic mucosa are not a subpopulation of the enterochromaffin cells, but constitute a distinct and independent cell type.

Immunohistochemical localization of the immunodominant differentiation antigen lacto-N-fucopentaose III in normal adult and fetal tissues.

Monoclonal antibodies were produced by immunizing rats with human small cell lung carcinoma (SCLC) cell lines. Monoclonal antibodies 600D11 and 624A12 were found to be directed against the ceramide pentasaccharide that contains the lacto-N-fucopentaose III (LNFP III) sequence of sugars, an isomer of the Lewis A blood group antigen. LNFP III is an immunodominant antigen whose reactivity is maintained in formalin-fixed paraffin-embedded sections (PS). LNFP III has been recognized in a number of human tumors including: SCLC; adenocarcinomas of the breast, gastrointestinal tract, genitourinary tract, and lung; renal cell carcinoma; neuroblastoma; and myelogenous leukemia. We now report the normal adult and fetal tissue distribution of the LNFP III antigen by immunoperoxidase staining on PS utilizing 600D11 and 624A12. Binding was demonstrated in bronchial epithelium and bronchial glands; squamous epithelium of the esophagus; gastric crypts, duodenal enterocytes and Brunners glands; argentaffin cells; jejunal and colonic goblet cells; pancreatic acinar cells; salivary glands; endocervical and exocervical cells; skin epidermis; myelinated motor fibers; cells of the adrenal medulla and anterior pituitary gland; polymorphonuclear leukocytes (PMNs); tissue macrophages and renal proximal tubules and loops of Henle. Staining was localized to cell membranes and within the cytoplasm, with greatest intensity at the apical and basal portions of the cells. These staining patterns were noted in adult and neonatal tissues, and initial expression could be traced to approximately the second trimester of fetal development. Knowledge of the normal tissue distribution of this immunodominant antigenic determinant may offer insight into its structural and functional role in benign and malignant tissues.

ENTEROCHROMAFFIN (ARGENTAFFIN) CELLS OF THE RAT GASTROINTESTINAL TRACT

Argentaffin cells from different parts of the gastro-intestinal tract were examined electron microscopically by the semi-thin/ultra-thin sectioning technique. All these cells contained round or pleomorphic secretory granules, which were mainly concentrated in the infranuclear zone, except in the antrum, where they occurred with a similar frequency in all the peri-nuclear areas. The highest frequency of granules was found in the colon. The largest granules were seen in the duodenum and the smallest in the large intestine. Argentaffin cells with processes oriented towards the glandular and villous surface were most frequent in the small intestine. No secretory granules were seen in the most apical part of these processes, where in an earlier study serotonin immunoreactivity was observed. This may indicate that serotonin can be stores in a non-granular form. No close topographic relationship was found between the argentaffin cells and other endocrine cell types or the nervous system.

Immunohistochemical localization of brain-gut hormones in gastric carcinoma with relation to argyrophil cells.

A total of 87 surgical cases of gastric carcinoma including 3 carcinoid tumors were investigated with the methods of silver reaction and immunoperoxidase stain for 8 different brain-gut hormones. Argyrophil (AP) cells were demonstrated in 38 cases (44%), argentaffin (AF) cells in 18 (21%) and endocrine cells in 13 (14%). The occurrence of endocrine cells had no relation with histological types. Glicentin cells were demonstrated in 10 cases, somatostatin in 7, motilin in 3, beta-endorphin in 2 and gastrin in one. Endocrine cells appeared generally in small numbers except one carcinoid tumor which had numerous somatostatin cells. No single cell positive for more than two kinds of hormones could be demonstrated. Two undifferentiated carcinomas looking like carcinoid tumors had argyrophil cells and endocrine cells of either somatostatin or beta-endorphin. These results suggest that carcinoid-like carcinoma or endocrine cell carcinoma may lie on the intermediate state between carcinoma and carcinoid tumor.

Gut endocrine cells in rat intestinal-tract carcinoma induced by 1,2-dimethylhydrazine.

Gut endocrine cells in a total of 122 intestinal-tract adenocarcinomas induced in inbred Wistar rats by 1,2-dimethylhydrazine dihydrochloride were examined histologically, ultrastructurally, and immunohistochemically for gastrin, somatostatin, vasoactive-intestinal polypeptide (VIP), and glicentin (enteroglucagon). Of the 122 tumors, argyrophil cells were detected in 42 tumors (34.3%) comprising 15 tumors of the well differentiated type and 27 tumors of the poorly differentiated type, including signet-ring-cell carcinomas. Of the 27 tumors of the poorly differentiated type, 12 were regarded as endocrine-cell carcinomas composed of numerous argyrophil or argentaffin cells and mucus-containing cells. Immunohistochemically, 7 of the 12 tumors had glicentin and two of these seven tumors also had gastrin and argentaffin cells synchronously. None of the tumors showed immunoreactivity for somatostatin and VIP. Nine of the 12 tumors metastasized to the lung, pancreas, liver, mesenterium, omentum, and lymph nodes. The metastatic foci of these tumors were also shown to have glicentin and argentaffin cells. Ultrastructurally, four types of endocrine granule were found in the tumor cells and amphicrine cells containing endocrine granules and mucous granules were noted. These endocrine-cell tumors were assumed to develop from totipotent immature cells of endodermal origin.

Histochemical, immunohistological and ultrastructural studies on neoplastic endocrine cells in carcinomas of the glandular stomach and small intestine of rats.

Neoplastic endocrine cells in the carcinomas of the stomach and small intestine of Wistar male rats, induced by MNNG or ENNG, were examined by histochemical, immunohistological and ultrastructural methods. Detection rates of argyrophil and argentaffin cells in the gastric cancers were 60% and 34.3%, respectively. MNNG induced small intestinal carcinomas contained argyrophil cells in 78.3% and argentaffin cells in 65.2%. Of 205 carcinomas of the small intestine induced by ENNG, 115 lesions (56.1%) contained argyrophil cells and 69 lesions (33.7%) contained argentaffin cells. In some of the cases, gastrin, glucagon and somatostatin were demonstrated in the tumor cells by the immunoperoxidase technique. Ultrastructurally, intracytoplasmic granules were observed, and a special form of endocrine cell which contained two different types of intracytoplasmic granules was also found (endocrine-exocrine cell). This latter finding indicates a multidifferentiating potentiality of tumor cell. Moreover, it would appear that these endocrine cells can arise from undifferentiated stem cells, and that the endocrine cells of the gastrointestinal tract are of the endodermal origin.

An immunohistochemical and ultrastructural study of Segi's cap, a large aggregation of gut endocrine cells, in bovine fetuses.

Segi's cap, a large aggregation of basal-granulated cells at the top of the intestinal villus, was studied in the proximal small intestine of bovine fetuses by histological, immunohistochemical and ultrastructural techniques. 1) Typical Segi's caps were seen in the duodenum and proximal jejunum in bovine fetuses. 2) Smaller groups of basal-granulated cells were found in the villous and partly also in the cryptal epithelium, as well as in the subepithelial lamina propria. The possible mechanism for their occurrence was discussed in connection with the fate of the cap. 3) Segi's caps were present in a neonatal calf before the suckling stage, but not in 3 or 4 week-old calves. The process of and reason for this abrupt disappearance of the caps are unknown. 4) The Segi's cap in bovine fetuses consisted mainly of argyrophil cells as demonstrated by Grimelius' and Hellerström-Hellman's silver methods. Only a few argentaffin cells were found in fetal caps using a modified Masson-Hamperl's silver method. 5) Immunohistochemically, somatostatin-, gastrin-, motilin- and secretin-immunoreactive cells were identified. 6) Four different endocrine cell types could be distinguished in the bovine Segi's cap on the basis of the ultrastructural appearance of their secretory granules.

Ultrastructural characteristics of the argentaffin cells. A study in the rat

Ultrastructural characteristics of the argentaffin cells. A study in the rat

Identification of the Serotonin-Synthesizing Endocrine Cells in the Rat Stomach by Electron Microscopic Radioautography and Amine Fluorescence

At least five morphological types of endocrine cells can be identified by electron microscopy in the rat stomach, most of which can decarboxylate 5-hydroxytryptophan to serotonin. To identify which o f these cells can synthesize 5-hydroxytryptamine from l-tryptophan, the physiological precursor of 5-hydroxytryptamine biosynthesis, pieces of rat stomach were incubated in organ culture with l-[ 3 H]tryptophan, with and without inhibitors, and were processed for light microscopic and electron microscopic radioautography. l-[ 3 H]tryptophan labeled all EC cells heavily, but labeled other endocrine cells-ECL, A-like, G, and D-only lightly. The labeling of EC cells was markedly reduced when 5-hydroxytryptamine synthesis was inhibited by either p-chlorophenylalanine or carbidopa, but was not appreciably reduced when protein synthesis was inhibited with cycloheximide. The labeling of Alike, G, and D cells by l-[ 3 H]tryptophan was similar to that produced by l-[ 3 H]leucine and was almost abolished by cycloheximide. The labeling of ECL cells by l-[ 3 H]tryptophan was shown by grain counts to be greater than the labeling of A-like and parietal cells, to persist after incubations with cycloheximide, but to be inhibited by p-chlorophenylalanine or carbidopa. Amine-fluorescence studies also indicated that ECL cells could synthesize some 5-hydroxytryptamine, and biochemical studies showed that the oxyntic mucosa could synthesize 5-hydroxytryptamine. Incubations of mucosa with 5-[ 3 H]hydroxytryptamine usually produced little or no labeling of endocrine cells. Thus, this study demonstrates that all EC cells can readily synthesize 5-hydroxytryptamine from tryptophan and therefore, as previously suggested from topographical and cytochemical studies, probably represent the argentaffin cells of light microscopy. This study also suggests that ECL cells can also synthesize and store some 5-hydroxytryptamine, but presumably not enough normally to be detected by conventional argentaffin, chromaffin, or amine-fluorescent techniques, which usually demonstrate few reactive cells in oxyntic glands, were ECL cells abound.

Ovarian Sertoli-Leydig cell tumors with heterologous elements. II. Cartilage and skeletal muscle: a clinicopathologic analysis of twelve cases.

Twelve ovarian Sertoli-Leydig cell tumors that contained heterologous elements in the form of skeletal muscle (nine cases), cartilage (seven cases) and neuroblastoma (one case) in either the primary of recurrent specimens are reported. Four of the primary tumors also contained foci of gastrointestinal type epithelium with argentaffin cells identifiable in two of them. The age of the patients ranged from 11-48 years (average, 24 years). Ten patients presented with an abdominal mass, one with abdominal pain and one with acute abdominal symptoms. Five of the patients, two of whom were virilized and one of whom was hirsute, had evidence establishing or suggesting androgen overproduction. All the tumors were unilateral. Four had ruptured preoperatively and two ruptured during the operation. The tumors averaged 18.5 cm in greatest diameter and had extensive areas of hemorrhage and necrosis in half the cases. On microscopic examination the Sertoli-Leydig cell component was poorly differentiated in 11 cases and of intermediate differentiation in one case. In two cases the primary tumor was a poorly differentiated Sertoli-Leydig cell tumor and heterologous elements were identified only in a recurrent mass. Follow-up of ten patients revealed that eight of them had died of tumor from five months to seven years postoperatively.

Ultrastructural and histochemical observations of neuroendocrine granules in nonneoplastic ovaries

Argyrophil and argentaffin cells are rarely seen in female genital tract structures. Their presence in normal, apparently healthy ovaries has not been reported until the present observation. Histochemical and ultrastructural studies demonstrated cells within the ovaries of two nonpregnant patients in the reproductive age that belong to the neuroendocrine or amine precursor uptake and decarboxylation (APUD) system. The lack of other discernible pathologic conditions suggests that these cells may be normal cellular constituents within the ovaries and lends support to the belief of a neural crest origin for primary ovarian carcinoid not associated with teratomatous or mucinous elements.

Induced fluorescence of serotonin in paraffin sections from carcinoid tumors

The detection of argentaffin cells and carcinoid tumor cells would be greatly facilitated if it were possible to apply methods of induced fluorescence directly on paraffin sections. We have subjected 5 μm thick sections from paraffin blocks containing carcinoids to different procedures for induced fluorescence. These include formaldehyde vapor at 80°C for 1 hour, 5% paraformaldehyde solution at room temperature followed by heating to 80°C for 15 min, and sucrose-phosphate buffer-glyoxylic acid solution either for 3 seconds at room temperature followed by heating to 80°C for 15 min or 15 min at 60°C in the solution. The different procedures were applied to sections which were non-deparaffinized, deparaffinized or deparaffinized and rehydrated to various degrees. With all procedures used we were able to induce histofluorescence of argentaffin cells and carcinoid tumor cells, and it was confirmed by microspectrofluorometry that the fluorophore originated from serotonin. Best results, when morphology was taken into consideration, were obtained with sections deparaffinized, transferred to 96% ethanol and then exposed to formaldehyde vapor.

Argyrophil cells in early gastric carcinoma: an immunohistochemical and ultrastructural study.

Eighteen argyrophil cell carcinomas in 101 early gastric carcinomas were explained histologically, ultrastructurally, and immunohistochemically for polypeptides, carcinoembryonic antigen (CEA), lysozyme, and human chorionic gonadotrophin (hCG). Seven of these 18 tumors had gastrin, and two of seven tumors also contained somatostatin. In all of these 18 tumors CEA were demonstrated. Seven had lysozyme and five of seven tumors also contained gastrin; hCG were present in four of the 18 tumors and two of four tumors had gastrin, CA, mucin, and lysozyme simultaneously. Argentaffin cells were found in seven of 18 tumors. Of the above seven tumors containing gastrin, three had argentaffin cells. Ultrastructurally, several types of secretory granules were noted and tumor cells resembling D1- or P cells were present in nine of the 18 tumors. Macroscopically, many of the tumors showed IIc or IIc + III type. Histologically, the 18 tumors consisted of six well differentiated adenocarcinomas and 12 poorly differentiated adenocarcinomas including signet-ring cell carcinoma. These 12 tumors frequently developed in the stomach of young females. In view of our previous investigations, it was suggested that the IIc-type argyrophil cell carcinoma histologically showing poorly differentiated adenocarcinoma may be related to scirrhous carcinoma of the stomach.

Scirrhous argyrophil cell carcinoma of the stomach with multiple production of polypeptide hormones, amine, CEA, lysozyme, and HCG.

Sixteen argyrophil cell carcinomas in 59 gastric scirrhous carcinomas were examined histologically, ultrastructurally, and immunohistochemically for polypeptide hormones, CEA, lysozyme, and HCG. In nine of these 16 tumors, polypeptides such as gastrin, somatostatin, and glucagon were demonstrated. Six of these nine tumors contained all three hormones, and three of these six tumors also had argentaffin cells. In all of these 16 tumors CEA were observed. Eight of them had CEA, lysozyme, and acid mucin synchronously. Of the above six tumors containing three peptides, three produced focal HCG. Ultrastructurally, several types of secretory granules were noted. Histologically, these 16 tumors showed poorly differentiated adenocarcinomas or signet ring cell carcinomas. Macroscopically, generalized type was 11 and localized type five. No hormonal syndrome was detected in any of the patients. It was suggested that these scirrhous argyrophil cell carcinomas of the stomach with the multifunction originate from totipotent immature cells of endodermal origin.

Analytical electron microscopic evaluation of the effects of paraformaldehyde pretreatment on the reaction of glutaraldehyde with biogenic amines.

Analytical electron microscopy was used to determine the quantitative effects of paraformaldehyde pretreatment on the formation of the biogenic amine-glutaraldehyde-chrome complex. Pretreatment with paraformaldehyde prevented the glutaraldehyde-chrome reaction with norepinephrine in the rat adrenal medulla. In contrast to the effect of paraformaldehyde on norepinephrine, pretreatment did not prevent the chrome reaction in serotonin-containing argentaffin cells of the gut. X-Ray energy spectrographic analysis revealed a significant decrease in chrome content in the paraformaldehyde treated tissue, but sufficient chrome did react to produce an electron-dense product. Thus by treating tissue with paraformaldehyde prior to the glutaraldehyde chrome procedure, serotonergic sites may be differentiated from catecholaminergic areas at the electron microscopic level.

Gastric carcinomas with argyrophil and argentaffin cells.

In series of 248 gastric carcinomas from surgical resections the occurence of argyrophil and argentaffin cells was studied. These cells were distinctly more frequent in diffuse than in tubular carcinomas. In all cases with both types of APUD-cells, argyrophil cells were more numerous than argentaffin cells. These tumors are considered to be a variant of current gastric carcinoma, with more pronounced APUD-differentiation of neoplastic cells. This variant stands apart from carcinoid tumors and also from those rare nasal adenocarcinomas with argentaffin cells.

Molecular forms of motilin in the mammalian and human gut and human plasma

Regional specific antibodies and gel permeation chromatography were used to analyze the molecular forms of motilin in the human gut and plasma and in the porcine gut. The same antibodies were also used for immunocytochemical localization of the cell of origin of motilin. Two main molecular forms of motilin were identified, peak I eluting at a volume suggesting a larger molecular weight substance (Kav = 0.16) and peak II coeluting with the porcine motilin standard (Kav = 0.67). Peak I accounted for over 50% of the total motilin immunoreactivity in the human gut, whereas in the porcine intestine it represented less than 5%. Peak I thus appeared to have a greater n-terminal sequence similarity with the porcine motilin standard. Chromatographic analysis of human plasma showed the presence of both immunoreactive peaks in approximately equal amounts which were both suppressed following infusion of somatostatin. Infusion of porcine motilin resulted in the expected rise of plasma immunoreactivity in the peak II position, whereas peak I was significantly depressed, suggesting the possibility of a negative feedback. Immunocytochemistry revealed that both antibodies immunostained equally the non argentaffin cell, which contained small round, electrondense granules. In addition some argentaffin enterochromaffin cells were stained predominantly with the n-terminal specific antibody.

Light and electron microscopic studies on the endocrine cells in the duck proventriculus.

The endocrine cells in the duck proventriculus were observed by light and electron microscopy. Most of argyrophil cells in the glandular lobules were bipolar or multipolar elongated in shape, but some were round in shape. In the superficial epithelium a few oval argyrophil cells were found. No argentaffin cell impregnated by Masson and Hamperl's method was found in the duck proventriculus. At least four types (I to IV) of endocrine cells were identified by ultrastructural morphology of the secretory granules and other cytoplasmic organelles. Type I cells were characterized by the presence of small (100-250 nm in diameter), spherical granules with various densities and internal structures. This type of cell seemed to correspond to the Grimelius- or Sevier-Munger-positive elongated cells. Type II cells contained numerous large, spherical (200-450 nm) granules and a few polymorphous (200-500 nm in longest diameter) granules with homogeneously electron-dense content surrounded by a limiting membrane. A few lipid-like droplets were found in their cytoplasm. Type III cells were characterized by the presence of medium-sized spherical granules (230-400 nm) of variable electron-density surrounded by limiting membranes. This type of cell seemed to correspond to the D cell in the duck pancreas. Type IV cells were distinguished by small, spherical granules (80-200 nm) with high-electron dense content surrounded by close-fitting membranes. All endocrine cells found in the duck proventriculus were of the closed type.