Changes in the mucosa of the duodenum and stomach as a result of experimental duodenal ulcers and vagotomy

By means of the transmissive and scanning electron microscopy methods and radioautography, structure of mucous membrane of the stomach and duodenum has been studied under experimentally induced duodenal ulcers before and after vagotomy during various time. The vagotomy results in accelerated healing of the ulcer defect. This is connected with an increased proliferative activity in the crypta cells, however, this is accompanied with deceleration of their differentiation. Under the duodenal ulcers the amount of chief and parietal cells increases in the gastric mucous membrane, this depends on gastrostasis produced by stenosis of the pylorus. At vagotomy the amount of the chief and parietal cells in the fundal glands of the mucous membrane decreases; this is accompanied with a lowered secretory activity.     

Morphological research on the effect of vagotomy on the parietal microflora of the stomach and duodenum

The effect of vagotomy on parietal microflora was studied in intact rats and rats with chronic experimental gastric ulcer using stereometry, as well as light, transmission and scanning electron microscopy. Vagotomy was found to increase the relative amount of parietal microflora on day 30 following denervation, especially in the duodenum and pylorus. Chronic gastric ulcers are also associated with a rise in the relative amount of parietal microflora.     

Quantitative ultrastructural studies on rat gastric zymogen cells under different physiological and experimental conditions

Summary Quantitative electron microscopical measurements performed on gastric zymogen cells were aimed at determinations of size and volume density of the zymogen granules, and size of cell and nuclear profiles. 17 groups of rats each comprising 6–8 animals were investigated: five of these groups were used to study the influence of fasting and feeding, three groups were killed at different intervals after a pilocarpine injection, and four other groups were investigated after an atropine injection. The remaining five groups of rats were operated on: vagotomy was performed on one group, vagotomy + pyloroplasty on another, pyloroplasty on the third group, and antrectomy on the last two groups. The operated rats were sacrificed 4 or 10 weeks after the operations.Pilocarpine was more effective than feeding in reducing the size and the amount of zymogen granules. After atropine the size and amount of zymogen granules tended to increase. Ten weeks after pyloroplasty, vagotomy + pyloroplasty, or antrectomy the mean size of the zymogen cells was reduced.Loss of trophic vagal impulses, duodenal regurgitation, and abnormal serum gastrin levels are factors which might be responsible for the zymogen cell hypotrophy in operated rats.Supported by grants from the Swedish Medical Research council (project no 12X-2298), M. Bergvalls Stiftelse, and Julins Fond. Excellent technical assistance was provided by Mrs. Sigrid Kilter and Miss Ulla Hedlund     

Duodenal glands in normal and vagotomized rats

In Wistar rats by means of the stereo- and morphometry methods, using semithin sections, histotopography of the duodenal glands (DG) and effect of vagotomy (VT) on them have been studied. The zone of the DG arrangement in the submucosal tela of the duodenum is about 5,000 mcm long. In the proximo-distal direction of the zone mentioned three parts can be distinguished--oral, middle and caudal. In the DG light-optically four varieties of mucocytes are identified. VT results in decreasing thickness of the zone of the DG arrangement, in outgrowth of the connective tissue and in a lowered amount of secrete in mucocytes.     

Cytodifferentiation of the chick pancreas. 3. The content and synthesis of ribonucleic acid and proteins in developing acinar cells

The content and synthesis of ribonucleic acid (RNA) and protein was studied by microphotometry and autoradiography in the developing pancreatic acinar cells of White Leghorn chick embryos. These findings were correlated with previously reported changes in ultrastructural components. Shortly before or concomitant with zymogen granulation, RNA synthesis increased, in association with increases in the amount of nucleolar and cytoplasmic protein. The cytoplasmic fraction was transitory, whereas the accumulated nucleolar protein was maintained and was soon followed by an increase in nucleolar RNA. Concomitantly, a decrease in chromosomal RNA was observed, with the total amount of nuclear RNA staying constant. When zymogen first appeared, nucleoli were greatly enlarged due to large amounts of RNA and protein; total cellular RNA and protein had decreased slightly, in association with a decrease in cell volume. Subsequent development presented smaller nucleoli with decreased amounts of RNA and protein. Total cellular RNA increased due to its accumulation in the cytoplasm, probably as ribosomes. The accumulation of zymogen and the enlargement of other cellular structures contributed to an increase in total cellular protein. Prior to hatching, total cell RNA and protein decreased in amount, probably due to a reduction in cell volume through cell division.     

Stereological analysis of the guinea pig pancreas. I. Analytical model and quantitative description of nonstimulated pancreatic exocrine cells

A stereological model which provides detailed quantitative information on the structure of the fasted, nonstimulated gland has been developed for the guinea pig pancreas. The model consists of morphologically defined space and membrane compartments which were used to describe the general composition of the tissue and the specific components of exocrine cells. The results are presented, where appropriate, relative to a cubic centimeter of pancreas, a cubic centimeter of exocrine cell cytoplasm, and to the volume of an average exocrine cell. The exocrine cells, accounting for 82% of the pancreas volume, consisted of 54% cytoplasmic matrix, 22% rough-surfaced endoplasmic reticulum (RER), 8.3% nuclei, 8.1% mitochondria, 6.4% zymogen granules, and 0.7% condensing vacuoles. Their total membrane surface area was distributed as follows: 60% RER, 21% mitochondria, 9.9% Golgi apparatus, 4.8% plasma membranes, 2.6% zymogen granules, 1.8% plasma membrane vesicles, and 0.4% condensing vacuoles. The application of this model to the study of membrane movements associated with the secretory process is discussed within the framework of an analytical approach.     

Function of coated membranes and multivesicular bodies during membrane regulation in stimulated exocrine pancreas cells

Summary Stimulation of secretion by pilocarpine results in a 70% loss of zymogen granules from pancreatic acinar cell during the first hr after injection of the drug. In previous work (Geuze and Poort, 1973), we found that the amount of membrane stored in the surface of the microvilli and of the numerous infoldings present in highly stimulated cells, increases during the first 2 hr and then decreases again during the 3rd hr after stimulation, concurrently with maximal endocytosis of sorbitol-[su14C].Further observations on the fine structure of stimulated cells at various time intervals after injection of pilocarpine showed that during the first hr numerous smooth vesicles and multivesicular bodies (mvb's) appear in the apical cytoplasm, while the number of coated vesicles and their relative total volume increase significantly 3 hr after stimulation.By infusion of ferritin in the pancreatic duct system in vivo and application of cytochemical techniques (osmium impregnation, electron microscope autoradiography and acid phosphatase cytochemistry) it could be established that after stimulated exocytotic secretion, redundant apical cell membrane is withdrawn by at least two routes: 1) During the initial rapid increase of the amount of apical cell membrane, withdrawal is accomplished by interiorization of luminal invaginations into smooth endocytotic vesicles, which in turn give rise to mvb's by infolding and subsequent fission of their limiting membrane. 2) Once the bulk of stored secretion granules has been discharged, endocytotic coated vesicles become gradually more prominent as carriers for redundant cell membrane. The contents of endocytotic structures ultimately become incorporated in residual bodies, suggesting lysosomal degradation of cell membrane prior to eventual reutilization.Coated vesicles also originate by pinching off from mature Golgi cisternae and condensing vacuoles. A possible function of the coated membranes in the concentration of exportable protein within forming secretory granules is discussed.     

Gastrodermal structure and feeding responses in the scleractinian Mycetophyllia reesi,a coral with novel digestive filaments

Mycetophyllia reesi Wells is a colonial scleractinian coral whose outer surface consists of a series of oral-pharyngeal openings that lack tentacles. The polyps also lack a column and cannot protrude from the colonial surface. Correspondingly, there is no central digestive cavity. Instead, the pharynx is directly connected to a series of radially arranged mesenterial ducts lying parallel to the skeleton. The ducts, composed primarily of ciliated cells with small mucus inclusions and large, compartmentalized mucocytes, house filaments that protrude through the oral apertures during feeding. The filaments may or may not be directly connected with or originate from the mesenterial ducts and are histologically distinct from them. They are therefore referred to as digestive, rather than mesenterial filaments. In contrast with other scleractinians, the digestive filaments are thin, unequally bilobed stalks with spatulate ends. The cnidoglandular (CG) lobe, the larger of the two, exhibits a distinct cellular zonation. Large mastigophore cnidae and elongated zymogen-like cells are clustered at its distal end. Neither of these cells appear to respond to particulate food material, suggesting that they may be employed in alternative modes of nutrition and/or competition. Behind the distal region, the CG lobe exhibits typical zymogen, mucus, and collar cells as well as numerous atrichous nematocysts. The atrichs and zymogen cells discharge during particulate feeding. Tracts of collar cells with particularly well-defined cilia, elongated rootlets, and mucus inclusions are found at the outer edge of the CG lobe. These cells disgorge their contents during feeding and appear to function in food transport. The smaller lobe of the filament is a muscular sheet containing well-defined fields of circular and longitudinal myofibrils along with associated neurons. Collar cells with lysosome-like inclusions and large, compartmentalized mucocytes are also characteristic of this region. There are no zooxanthellae in the filaments, but these endosymbionts are present as a thin layer in the oral-most portion of the gastrodermis. The cellular zonation and multi-functionality of these digestive filaments suggest another example of a cnidarian structure at the organ level of complexity.     

Effects of gastric mucosa irradiation with helium-neon laser on epithelial cells

Effect of various doses of low-tense laser irradiation on proliferative and functional morphology of fundal gastric epitheliocytes in rats was studied by endogastric irradiation with the methods of histochemistry, autoradiography, transmission and scanning electron microscopy. It was shown that laser irradiation effect varied with its dose and was spread on gastric mucosa lying at the site of direct influence. The doses of irradiation that had no alternative effect induced reconstruction of epitheliocytes especially mucocytes indicating intensification of specific function in them, as well as an increase of proliferative activity.     

Cytological effects of experimental exposure to Hg on the gill epithelium of the European flat oyster Ostrea edulis: ultrastructural and quantitative changes related to bioaccumulation

The significance of ultrastructural changes in the gill epithelial cells as a parameter of detection of Hg exposure in the flat oyster, Ostrea edulis, was tested by a 34-day exposure to 5 microg l(-1) of Hg. The concentration of Hg (38.76 microg g(-1) dry weight) in gill tissue was maximal after 25 days and then decreased. The histological pattern of gill filaments in control samples did not vary throughout the experiment, except for the volume of mucocytes after 4 days of exposure, as an adaptation to experimental conditions. This volume increased significantly and then decreased according to the accumulation of Hg in the gills. After 18 days, absorptive and ciliated cells of the gill epithelium showed blebs in microvilli membranes, discocilia and swollen mitochondrial cristae. Both cell types showed distinct cellular lysis stages after 25 days of exposure. These are the target cells of Hg and other metals and the reported hypertrophy of mucocytes increase occurs in response to pollution by Hg, which could contribute to the detoxification process.     

Morphometry and elemental analysis of rat exocrine pancreas following administration of trypsin inhibitor

The morphological responses of the exocrine pancreas of the adult male rat to soybean trypsin inhibitor (STI) were studied by ultrastructural morphometry and electron probe X-ray microanalysis. STI administered orally in drinking water for 14 days resulted in a 72% increase in the wet weight of the pancreas. This enlargement was due, largely, to an increase in acinar cell mass. Volume increases in the acinar cell mass and extra-acinar cell compartment were 72 and 30%, respectively. The estimated total number of acinar cells in the mean exocrine pancreas was 500 million in the control and 630 million in the experimental group, representing an increase of 27%. Acinar cell volume was 1,790 microns 3 for the control and 2,457 microns 3 for the STI group. The pronounced morphometric changes of the organelles in the STI group were: the mean nucleolar volume increased by 56%; the volume of zymogen granular mass per cell increased by 93%; the volume of the Golgi complex and the condensing vacuoles per cell increased by 52 and 100%, respectively, whereas the membrane area of the Golgi complex and the condensing vacuoles increased by 98 and 47%, respectively. Spectral analysis of seven elements (Na, Mg, P, S, Cl, K and Ca) showed significant changes for nuclei, zymogen granules and mitochondria following STI: nuclei showed Na, P, K increased; zymogen granules showed Na, P, S, K increased, Cl decreased; mitochondrial particles showed Mg, P, Cl, Ca increased, and the mitochondrial matrix showed S decreased. The persistent uptake of STI probably resulted in a continual release of a trophic hormone acting on pancreatic tissue components, consequently causing hyperplasia and hypertrophy of the exocrine pancreas to accommodate a heightened demand for synthesis of exportable proteins.     

Ultrastructural changes in rabbit heart mitochondria during the perinatal period: Neonatal transition to aerobic metabolism

Ultrastructural changes in mitochondria of rabbit left ventricular myocardial cells were measured during the interval from 3 days before to 4 days after birth. This interval is characterized by a transition from partially anaerobic to aerobic metabolism and by a concomitant increase in the work of contraction. Measurements were made by recently developed morphometric techniques [Smith, H. E., and Page, E. (1976). J. Ultrastruct. Res.55, 31–41]. It was shown that the perinatal transition was associated with a rapid and large accumulation of mitochondria and myofibrils, reflected by increases of 29–35% and 37–41% in the mitochondrial and myofibrillar fractions of cell volume. The mitochondria thus accumulated were packed more densely with cristae. The membrane area of cristae + inner membrane per unit of mitochondrial volume rose from an antenatal value of 47 μm²/μm³ to a peak of 64 μm²/μm³ 2 days after birth, then declined significantly. The relative and absolute increases in the membrane area and volume of respiratory membrane were associated with a relative decrease in matrix volume. These measurements also permitted the calculation of the area of mitochondrial respiratory membrane per unit myofibrillar volume. This physiologically important index, which relates the amount of ATP-producing membrane to the volume of the principal ATP-consuming organelle, increased progressively throughout the perinatal period.     

A morphometric study of developing pancreatic acinar cells of rats during prenatal life

Summary The pancreatic acinar cells of rat embryos obtained at 15, 17, 19 and 21 days of gestation have been examined using fine-structural and morphometric techniques.Morphometric analysis demonstrates significant variations in the average volume of the cell, nucleus and cytoplasm, and the volume, surface and numerical densities of various cytoplasmic organelles during fetal life. In particular, the volume and surface densities of rER exhibit maximal values at 19 days of gestation, suggesting that secretory proteins are produced most actively at this time. Further-more, membrane continuity between the nuclear envelope and rER is frequently discernible in acinar cells, indicating that at this stage the rER is mainly derived from the nuclear envelope. Zymogen granules first appear at 17 days of gesstation. By 21 days, they occupy the greater part of the cytoplasm of the acinar cells, no polarity being seen in their distribution pattern. No direct evidence for the secretion of zymogen granules has been observed during fetal life.It therefore appears that membrane transport involved with intracellular movement of newly synthesized proteins from rER via the Golgi complex to zymogen granules occurs in one direction and lacks regulation.     

Concanavalin a binding and cytodifferentiation in pancreatic acinar carcinoma of rat

The binding of concanavalin A to the plasmalemma of acinar carcinoma cells was characterized by electron microscopy utilizing horseradish peroxidase. Heavy labeling due to specific concanavalin A binding was detected on the plasmalemma of undifferentiated carcinoma cells lacking zymogen maturation, neoplastic cells of intermediate differentiation with only occasional zymogen granules, and highly differentiated acinar carcinoma cells containing numerous cytoplasmic zymogen granules. The plasmalemma of acinar carcinoma cells was also compared to the normal pancreatic acinar cell plasmalemma by measurement of specific ¹²⁵I-labeled concanavalin A binding. Although only about one-third of pancreatic acinar carcinoma cells demonstrate mature zymogen differentiation, the acinar carcinoma had a full complement of normal plasmalemma receptors for ¹²⁵I-labeled concanavalin A. It is concluded that, unlike normal pancreas, the presence of concanavalin A receptors on the plasmalemma of acinar carcinoma cells is not a specific membrane marker for differentiated cells containing zymogen granules.     

Topographical and planar distribution of Helix pomatia lectin-binding glycoconjugates in secretory granules and plasma membrane of pancreatic exocrine acinar cells of the rat: demonstration of membrane heterogeneity

The lectin-gold technique was used to detect Helix pomatia lectin (HPL) binding sites directly on thin sections of rat pancreas embedded in Lowicryl K4M and on freeze-fractured preparations of rat pancreas submitted to fracture label. On thin sections of acinar cells, whereas the content of zymogen granules was negative or weakly labeled, the limiting membrane displayed a high degree of labeling. In the Golgi complex, labeling by HPL was localized on the trans saccules and the limiting membrane of the condensing vacuoles. The latter appeared to be more intensely labeled than the membrane of the zymogen granules. Intense labeling by HPL was also observed along the microvilli and the plasma membrane. In contrast to the weak labeling of the zymogen-granule content, labeling of the acinar lumen was intense. Fracture-label preparations revealed preferential partition of HPL-binding sites to the exoplasmic half of the zymogen-granule and plasma membranes. The population of zymogen granules was, however, heterogeneous with respect to labeling intensity; the exoplasmic fracture-face of the plasma membrane was intensely and uniformly labeled, while the protoplasmic membrane halves were only weakly labeled. These observations were further confirmed and extended by the thin-section fracture-label approach. In addition, favorable profiles of thin sections of freeze-fractured zymogen granules showed that the labeling was not associated with the external surface of the limiting membrane, but rather localized over the exoplasmic fracture-face. We conclude that 1) zymogen granules contain little HPL-binding glycoconjugates, 2) HPL-binding sites are preferentially associated with the exoplasmic half of the zymogen-granule and plasma membranes, and 3) the limiting membrane of the immature condensing vacuoles carries a greater number of HPL-binding sites than that of the mature zymogen granules. These last, in turn, constitute a heterogenous population with respect to labeling density. These results support the current view that glycoconjugates are directed toward the lumen in secretory granules but become external to the cell surface after fusion of the secretory-granule membrane with the plasma membrane. Also, the results reflect membrane modifications during the maturation process of secretory granules in the exocrine pancreas in which glycoproteins are removed from the limiting membrane of the granule to become soluble and secreted with the content.     

Apactin is involved in remodeling of the actin cytoskeleton during regulated exocytosis

Apactin is an 80-kDa type I membrane glycoprotein derived from pro-Muclin, a precursor that also gives rise to the zymogen granule protein Muclin. Previous work showed that apactin is efficiently removed from the regulated secretory pathway and targeted to the actin-rich apical plasma membrane of the pancreatic acinar cell. The cytosolic tail (C-Tail) of apactin consists of 16 amino acids, has Thr casein kinase II and Ser protein kinase C phosphorylation sites, and a C-terminal PDZ-binding domain. Secretory stimulation of acinar cells causes a decrease in Thr phosphorylation and an increase in Ser phosphorylation of apactin. Fusion peptides of the C-Tail domain pulldown actin, ezrin, and EBP50/NHERF in a phosphorylation-dependent manner. HIV TAT-C-Tail fusion peptides were used as dominant negative constructs on living pancreatic cells to study effects on the actin cytoskeleton. During secretory stimulation, TAT-C-Tail-Thr/Asp phosphomimetic peptide caused an increase in actin-coated zymogen granules at the apical surface, while TAT-C-Tail-S/D phosphomimetic peptide caused a broadening of the actin cytoskeleton. These data indicate that stimulation-mediated Thr dephosphorylation allows decreased association of apactin with EBP50/NHERF and fosters actin remodeling to coat zymogen granules. Stimulation-mediated Ser phosphorylation increases apactin association with the actin cytoskeleton, maintaining tight bundling of actin microfilaments at the apical surface. Thus, apactin is involved in remodeling the apical cytoskeleton during regulated exocytosis in a manner controlled by phosphorylation of the apactin C-Tail.     

Histology and ultrastructure of the salivary glands in Bulla striata (Mollusca, Opisthobranchia)

Abstract. The ribbon‐shaped salivary glands in Bulla striata were studied with light microscopy and transmission electron microscopy (TEM). Secretion is produced in tubules formed by two types of secretory cells, namely granular mucocytes and vacuolated cells, intercalated with ciliated cells. A central longitudinal duct lined by the same cell types collects the secretion and conducts it to the buccal cavity. In granular mucocytes, the nucleus is usually central and the secretory vesicles contain oval‐shaped granular masses attached to the vesicle membrane. Glycogen granules can be very abundant, filling the space around the secretory vesicles. These cells are strongly stained by PAS reaction for polysaccharides. Their secretory vesicles are also stained by Alcian blue, revealing acidic mucopolysaccharides, and the tetrazonium reaction detects proteins in minute spots at the edge of the vesicles, corresponding to the granular masses observed in TEM. Colloidal iron staining for acidic mucopolysaccharides in TEM reveals iron particles in the electron‐lucent region of the vesicles, while the granular masses are free of particles. In vacuolated cells, which are thinner and less abundant than the granular mucocytes, the nucleus is basal and the cytoplasm contains large electron‐lucent vesicles. These vesicles are very weakly colored by light microscopy techniques, but colloidal iron particles could be observed within them. The golf tee‐shaped ciliated cells contain some electron‐dense lysosomes in the apical region. In these cells, the elongated nucleus is subapically located, and bundles of microfibrils are common in the slender cytoplasmic stalk that reaches the basal lamina. The morphological, histochemical, and cytochemical data showed some similarities between salivary glands in B. striata and Aplysia depilans. These similarities could reflect the phylogenetic relationship between cephalaspidean and anaspidean opisthobranchs or result from a convergent adaptation to an identical herbivorous diet.     

Mucocytes with micronuclei and sowing with the coccoid forms of <Emphasis Type="Italic">Helicobacter pylori</Emphasis> in a mucous membrane of human stomach

In accordance with the solution of IARC, the Helicobacter pylori (H. pylori) refers to carcinogens of the first group. As the carcinogenic factors have a mutagen effect, we have undertaken the cytogenetic testing of 62 patients with chronic nonatrophic gastritis (40 of which have H. pylori-associated gastritis) by account of the micronuclei in mucocytes of tectorial-pit epithelium of the mucous membrane of the antral region of the stomach. The detection of H. pylori cells in the mucous membrane of the stomach (SMM) was performed with the help of immunocytochemical method that permitted us to visualize both the bacillar and coccoid forms, as well as to evaluate the degree of sowing of SMM with the coccoid forms of H. pylori. In the patient group with H. pylori-associated gastritis, the frequency rate of mucocytes with micronuclei in SMM appears to be considerably higher than in the group of patients whose SMM was not infected with H. pylori (P < 0,05). A high scale of sowing with the coccoid forms of H. pylori was accompanied by a significantly heightened level of mucocytes with micronuclei in the SMM. In connection with this and on the basis of modern notions of carcinogenesis, based on mutagen modifications in somatic cells, patients that exhibit high sowing with coccoid forms of H. pylori may be placed in the group of heightened oncologic hazards.