scholarly journals An Electron Microscopic Study of the Intestinal Villus

1959 ◽  
Vol 5 (3) ◽  
pp. 373-384 ◽  
Author(s):  
Sanford L. Palay ◽  
Leonard J. Karlin

The intestinal pathway for absorbed fat was traced in thin sections of intestinal villi from rats fed corn oil by stomach tube after a fast of 24 to 40 hours. For electron microscopy the tissues were fixed in chilled buffered osmium tetroxide and embedded in methacrylate. For light microscopy, other specimens from the same animals were fixed in formal-calcium, mordanted in K2Cr2O7, and embedded in gelatin. Frozen sections were stained with Sudan black B or Sudan IV. About 20 minutes after feeding, small fat droplets (65 mµ maximal diameter) appear in the striated border between microvilli. At the same time fat particles are seen within pinocytotic vesicles in the immediately subjacent terminal web. In later specimens the fat droplets are generally larger (50 to 240 mµ) and lie deeper in the apical cytoplasm. All intracellular fat droplets are loosely enveloped in a thin membrane, the outer surface of which is sometimes studded with the fine particulate component of the cytoplasm. This envelope, apparently derived from the cell surface by pinocytosis, has at this stage evidently become a part of the endoplasmic reticulum. Just above the nucleus numerous fat droplets lie clustered within the dilated cisternae of the Golgi complex. As absorption progresses fat droplets appear in the intercellular spaces of the epithelium, in the interstitial connective tissue spaces of the lamina propria, and in the lumen of the lacteals. All of these extracellular fat droplets are devoid of a membranous envelope. The picture of fat absorption as reconstructed from these studies involves a stream of fat droplets filtering through the striated border, entering the epithelial cell by pinocytosis at the bases of the intermicrovillous spaces, and coursing through the endoplasmic reticulum to be discharged at the sides of the epithelial cell into extracellular spaces. From the epithelial spaces, the droplets move into the lamina propria and thence into the lymph. If the lumen of the endoplasmic reticulum is considered as continuous with the extracellular phase, then the entire pathway of fat absorption may be regarded as extracellular. However, it is impossible to evaluate from the electron microscopic evidence thus far available the quantitative importance of particulate fat absorption by the mechanism described.

1983 ◽  
Vol 31 (8) ◽  
pp. 987-999 ◽  
Author(s):  
J Roth

A method is described for the electron microscopic detection of lectin-binding sites in different cellular compartments and extracellular structures that uses thin sections from resin-embedded tissues. Various lectins (Ricinus communis lectin I and II, peanut lectin, Lotus tetragonolobus lectin, Ulex europeus lectin I, Lens culinaris lectin, Helix pomatia lectin, and soybean lectin) were bound to particles of colloidal gold and used for direct staining of thin sections or glycoprotein--gold complexes were prepared and applied in an indirect technique (concanavalin A and horseradish peroxidase--gold complex; wheat germ lectin and ovomucoid--gold complex). The details for preparation of such complexes from 14 nm gold particles are reported. The conditions of tissue processing that gave satisfactory staining results and good fine structure preservation were mild aldehyde fixation without osmification and low temperature embedding with the hydrophilic resin Lowicryl K4M. None of the so-called etching procedures was necessary prior to labeling of Lowicryl K4M thin sections. Examples of the use of this approach for detection of glycoconjugates in the rough endoplasmic reticulum, Golgi apparatus, and mucin of intestinal goblet cells as well as plasma membrane and various intracellular structures of absorptive intestinal and renal tubular cells are shown. A comparison is made with preembedding staining results on Concanavalin A-binding site localization in rat liver which shows that problems of penetration common in such a technique are circumvented by the postembedding approach described here. Concanavalin A-binding sites were not only consistently found in nuclear envelope, rough and smooth endoplasmic reticulum, plasma membranes, and collagen fibers, but also in mitochondria, glycogen, ribosomes, and nucleus. These data and those of a previous investigation (Roth J, Cytochem 31:547, 1983) prove the applicability of this cytochemical technique for postembedding localization of glycoconjugates by light and electron microscopy.


Parasitology ◽  
1957 ◽  
Vol 47 (1-2) ◽  
pp. 66-69 ◽  
Author(s):  
H. Meyer ◽  
I. De Andrade Mendonça

Toxoplasma ‘Nicolle et Manceaux’ has been examined in the electron microscope in thin sections of infected tissue cultures. The intracellular forms, single or in rosette formations, show that the parasite has no flagella, cilia or other organelles for locomotion. It is covered by a double membrane which is especially well denned at the two poles. A ring-like formation, often slightly protruding, can be seen at one end of the parasite, and in connexion with this, long, homogeneous, dark-stained inclusions of still unknown nature. At the opposite pole a distinct opening has been found.Mitochondria and strands of an endoplasmic reticulum are also present in the cytoplasm and a fine striation in the region of the periplast. No dividing forms have been observed. Near the nucleus, in the region of what may correspond to the centrosome and the Golgi apparatus, a few extremely fine granules and a spiral-like formation of fine striae or tubules have been observed.This work has been partly supported by the National Research Council of Brazil. The cost of reproduction of figures was defrayed by the Institute de Biofísica da Universidade do Brasil.


1961 ◽  
Vol 11 (2) ◽  
pp. 333-347 ◽  
Author(s):  
Susumu Ito

An electron microscopic survey has been made of the gastric parietal or oxyntic cell of the human, cat, beaver, dog, hamster, rat, mouse, and bat, and of the corresponding cell type in two species of frog, two species of toad, and the horned lizard. A feature consistently found in the parietal cells of the mammals or their equivalent in the lower vertebrates is the agranular endoplasmic reticulum, which takes the form of branching and anastomosing small tubules approximately 200 to 500 A in diameter, sometimes expanded into flattened cisternae. In mammalian parietal cells this form of the endoplasmic reticulum is found only in limited amounts, but in the corresponding secretory cells of the amphibia and reptilia the tubular agranular reticulum is abundant. It is believed to comprise a more or less continuous system of channels, but owing to their tortuous course only short profiles are seen in thin sections. Immediately subjacent to the plasmalemma at the free surface, the cytoplasm is relatively free of organelles but is occasionally traversed by the agranular reticulum, which appears to be continuous at some points with the cell surface. The possible participation of the agranular endoplasmic reticulum in hydrochloric acid secretion is discussed.


Author(s):  
M. R. Edwards ◽  
W. A. Samsonoff

Papillomas in catfishes have been described (1) but the presence of viruses in these tumors has not (2). This report is concerned with the study of a papilloma found on the lower lip of a brown bullhead (letalurus nebulosus) which had been frozen prior to arrival at our laboratory.Tumor tissues were thawed in 2.5% glutaraldehyde and postfixed in 1$ osmium tetroxide. Both fixatives were prepared in Millonig's buffer. Fixation, washing, dehydration, and Epon embedding were accomplished according to conventional electron microscopic techniques.Examination of thin sections revealed virus-like particles in epidermal cells which constituted the major part of the neoplasm (Fig. 1). No particles were found in the connective tissue surrounding the epidermal papillae. The cells containing particles were usually isolated from one another, had a spindle or fusiform shape, and exhibited many cytoplasmic extensions in random directions. Their nuclei were pleomorphic and displayed irregular nuclear envelopes with relatively large lacunae. Vesiculation of the cytoplasm was extensive, apparently caused by dilations of the endoplasmic reticulum.


1982 ◽  
Vol 92 (1) ◽  
pp. 92-107 ◽  
Author(s):  
D Louvard ◽  
H Reggio ◽  
G Warren

Rabbits were immunized with membrane fractions from either the Golgi complex or the rough endoplasmic reticulum (RER) by injection into the popliteal lymph nodes. The antisera were then tested by indirect immunofluorescence on tissue culture cells or frozen, thin sections of tissue. There were may unwanted antibodies to cell components other than the RER or the Golgi complex, and these were removed by suitable absorption steps. These steps were carried out until the pattern of fluorescent labeling was that expected for the Golgi complex or RER. Electron microscopic studies, using immunoperoxidase labeling of normal rat kidney (NRK) cells, showed that the anti-Golgi antibodies labeled the stacks of flattened cisternae that comprise the central feature of the Golgi complex, many of the smooth vesicles around the stacks, and a few coated vesicles. These antibodies were directed, almost entirely, against a single polypeptide with an apparent molecular weight of 135,000. The endoplasmic reticulum (ER) in NRK cells is an extensive, reticular network that pervades the entire cell cytoplasm and includes the nuclear membrane. The anit-RER antibodies labeled this structure alone at the light and electron microscopic levels. They were largely directed against four polypeptides with apparent molecular weights of 29,000, 58,000, 66,000, and 91,000. Some examples are presented, using immunofluorescence microscopy, where these antibodies have been used to study the Golgi complex and RER under a variety of physiological and experimental condition . For biochemical studies, these antibodies should prove useful in identifying the origin of isolated membranes, particularly those from organelles such as the Golgi complex, which tend to lose their characteristic morphology during isolation.


1959 ◽  
Vol 5 (3) ◽  
pp. 363-371 ◽  
Author(s):  
Sanford L. Palay ◽  
Leonard J. Karlin

The structure of the intestinal villus of the rat was studied in thin sections of tissue fixed in buffered osmium tetroxide and embedded in methacrylate. The simple columnar epithelium investing the villus is surmounted by a striated border consisting of slender projections of the cell surface. These microvilli are arranged in almost crystalline, hexagonal array, and increase the apical surface area of the cell by a factor of 24. The core of each microvillus is filled with fine fibrils which arise from the filamentous substance of the terminal web underlying the striated border. Each microvillus is covered by a tubular extension of the plasma membrane of the epithelial cell. Pinocytotic vesicles originating from the plasma membrane occur at the bases of the intermicrovillous spaces. The nucleus, mitochondria, and the endoplasmic reticulum of the epithelial cell display no unusual features. Small bits of ergastoplasm occur in the apical cytoplasm. A thin basement membrane separates the epithelium from the lamina propria which consists of vessels, nerves, and numerous lymphocytes, eosinophiles, mast cells, plasma cells, smooth muscle fibers, and macrophages suspended in a delicate stroma of fibroblasts and collagen fibers. Intercellular fat droplets often occur in this stroma, even in animals fasted for 40 hours. The blood capillaries are distinguished by their extremely attenuated, fenestrated endothelial cells. The lacteal has a thicker endothelium which, although not fenestrated, appears to have significant interruptions, especially at the margins between neighboring lining cells. Strands of smooth muscle always accompany the lacteal but do not form an integral part of its wall. Unmyelinated nerves, many of which are too small to be distinguished with the light microscope, course through the lamina propria in association with the vessels. The nerve fibers evidently do not cross the basement membrane into the epithelium. Neuromuscular junctions or other terminal apparatus were not found.


Author(s):  
V. F. Allison ◽  
G. C. Fink ◽  
G. W. Cearley

It is well known that epithelial hyperplasia (benign hypertrophy) is common in the aging prostate of dogs and man. In contrast, little evidence is available for abnormal epithelial cell growth in seminal vesicles of aging animals. Recently, enlarged seminal vesicles were reported in senescent mice, however, that enlargement resulted from increased storage of secretion in the lumen and occurred concomitant to epithelial hypoplasia in that species.The present study is concerned with electron microscopic observations of changes occurring in the pseudostratified epithelium of the seminal vescles of aging rats. Special attention is given to certain non-epithelial cells which have entered the epithelial layer.


Author(s):  
J. R. Ruby

Parotid glands were obtained from five adult (four male and one female) armadillos (Dasypus novemcinctus) which were perfusion-fixed. The glands were located in a position similar to that of most mammals. They extended interiorly to the anterior portion of the submandibular gland.In the light microscope, it was noted that the acini were relatively small and stained strongly positive with the periodic acid-Schiff (PAS) and alcian blue techniques, confirming the earlier results of Shackleford (1). Based on these qualities and other structural criteria, these cells have been classified as seromucous (2). The duct system was well developed. There were numerous intercalated ducts and intralobular striated ducts. The striated duct cells contained large amounts of PAS-positive substance.Thin sections revealed that the acinar cells were pyramidal in shape and contained a basally placed, slightly flattened nucleus (Fig. 1). The rough endoplasmic reticulum was also at the base of the cell.


Author(s):  
F. G. Zaki

Addition of lithocholic acid (LCA), a naturally occurring bile acid in mammals, to a low protein diet fed to rats induced marked inflammatory reaction in the hepatic cells followed by hydropic degeneration and ductular cell proliferation. These changes were accompanied by dilatation and hyperplasia of the common bile duct and formation of “gallstones”. All these changes were reversible when LCA was withdrawn from the low protein diet except for the hardened gallstones which persisted.Electron microscopic studies revealed marked alterations in the hepatic cells. Early changes included disorganization, fragmentation of the rough endoplasmic reticulum and detachment of its ribosomes. Free ribosomes, either singly or arranged in small clusters were frequently seen in most of the hepatic cells. Vesiculation of the smooth endoplasmic reticulum was often encountered as early as one week after the administration of LCA (Fig. 1).


Author(s):  
J. T. Stasny ◽  
R. C. Burns ◽  
R. W. F. Hardy

Structure-functlon studies of biological N2-fixation have correlated the presence of the enzyme nitrogenase with increased numbers of intracytoplasmic membranes in Azotobacter. However no direct evidence has been provided for the internal cellular localization of any nitrogenase. Recent advances concerned with the crystallizatiorTand the electron microscopic characterization of the Mo-Fe protein component of Azotobacter nitrogenase, prompted the use of this purified protein to obtain antibodies (Ab) to be conjugated to electron dense markers for the intracellular localization of the protein by electron microscopy. The present study describes the use of ferritin conjugated to goat antitMo-Fe protein immunoglobulin (IgG) and the observations following its topical application to thin sections of N2-grown Azotobacter.


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