Search

An antiserum prepared against lysozyme isolated from mucosal scrapings of mouse small intestine was used to stain sections of mouse small intestine with the indirect fluorescent antibody technique. Mucosal fluorescence was confined to the base of the crypts of Lieberkuhn, where Paneth cells are located. After the intravenous administration of 4 mg of pilocarpine fluorescence was no longer found in the Paneth cell but in the crypt lumen. Perfusion studies confirmed these findings. The basal lysozyme output of 0-1 to 0-4 mug/ml was raised to peak rates of 1-8 to 6-5 mug/ml after the intravenous administration of 1 mg of pilocarpine. Our results demonstrate that the lysozyme of the succus entericus is, at least in part, derived from the Paneth cell, and is probably present in the Paneth cell granules. Its secretion is stimulated by pilocarpine. Our model could be very useful for studying the function of the Paneth cell, which probably forms part of an intestinal defence system.

Photomicrographic evidence, mitotic figures and 3H-thymidine labeling among Paneth cells, is presented in support of the hypothesis that Paneth cells are replaced by cells proliferating in the bottom of the intestinal crypt rather than by migration of cells from the mid-crypt region.

At the ultrastructural level we have examined the interactions between the intestinal flagellate, Hexamita muris, and the epithelial cells of the duodenal crypts where trophozoites of this parasite were particularly concentrated in infected rats. Both structurally intact and partially digested trophozoites of Hexamita were commonly found within digestive vacuoles in Paneth cells, but were never encountered intracellularly in any other cell type. Although up to eight intact trophozoites were observed in vacuoles within a single Paneth cell, no pathological effects on the Paneth cells were noted. Varying stages in the digestion of trophozoites were observed and because of the complex structure of the trophozoites, especially the resistance of flagellar microtubules to digestion, it was possible to reconstruct the evolution of residual bodies from phagolysosomes. These results indicate that Paneth cells can function as fixed phagocytes and suggest, therefore, that they play a role in controlling the intestinal flora in the crypt of Lieberkuhn.

The secretion of Paneth cells in the rat has been stimulated by pilocarpine, the small intestinal secretions have been collected and their free amino acid concentrations measured. Comparison with a control group of animals not given pilocarpine showed a statistically significant increase in the free amino acid concentrations. 65Zn has been used to label Paneth cells in experiments which indicate that this increase in luminal amino acid concentrations is the result of the action of pilocarpine on the Paneth cell. Experiments using tritiated thymidine indicate that pilocarpine does not influence the rate of cell turnover in the rat small intestine. We conclude that the Paneth cell secretion is composed of amino acids.

The histochemistry of mouse Paneth cell granules was investigated with alcian blue (AB, pH 1.0 or 2.5), periodic acid-Schiff reagent (PAS), high iron diamine (HID) and Biebrich scarlet (pH 9.5). After formalin fixation, an AB pH 2.5-PAS sequence demonstrates alcianophilic rims surrounding weakly PAS-reactive granule cores; the HID-AB and AB pH 1.0-PAS sequences show peripheral staining indicative of a sulfomucin in most granules and a carboxymucin in some granules, the latter predominantly in the distal small intestine. When stained with PAS alone, rims of Paneth granules stain intensely and cores weakly. After HgC12 fixation, HID staining indicative of sulfomucin occurs in the cores of most granules and in the rims of many granules. An AB-Biebrich scarlet sequence shows acid mucosubstance in the rims and strongly basic protein (presumably arginine rich) in the cores of the granules. The basic protein-rich cores are larger toward the bottom of the crypts and toward the distal small intestine. The acid mucosaccharide-rich rims appear thinner surrounding the larger granules at the bottom of the crypts. Fine structural studies have demonstrated dialyzed iron staining indicative of acid mucosubstance in the rims of mouse Paneth cell granules and in small profiles and some lamellae of the Golgi zone.

Paneth cells from fasted, fed and pilocarpine stimulated mice were studied with the light and electron microscopes. While human and rat Paneth cells contain structurally homogeneous granules, most mouse Paneth cell granules were found to be structurally heterogeneous both by light and electron microscopy. These structurally heterogeneous granules consisted of an outer rim or halo which surrounded the central core of the granule. These halos varied strikingly in electron density ranging from much paler to much darker than the granule core. Electron density of the halos correlated well with their affinity for toluidine blue in light microscopic preparations. The granule halo could be stained with both alcian blue and with PAS indicating that it contained acid mucopolysaccharide while the central core of the granule could be stained only with PAS and not with alcian blue. While actively secreting Paneth cells were seen in preparations from fasting mice, the secretory activity was increased after feeding and after stimulation with pilocarpine. The actively secreting mouse Paneth cells discharged their granules into the crypt lumen by merocrine secretion. The fine structure of mouse Paneth cell granules was the same in resting and in actively secreting cells.

Nach spezifischer i.v. Injektion wird gefunden, dass Dopamin und 3,4-Dioxyphenylalanin-aminosaure im Zytoplasma der Panethschen Zellen des Mauseduodenums angehauft sind. Wahrend Niamid® das meist rasche Verschwinden der Formaldehyd-verursachten Aminfluoreszenz in den Granula von Panethschen Zellen verzogerte, blieb Reserpin wirkungslos.

Salivary, gastric and pancreatic juice, bile, goblet cells and enterocytes contribute to intestinal calcium excretion. Additionally, the light microscopic GBHA technique (KASHIWA, 1966) drew attention to the Paneth cells. These excretory cells are found predominantly in the ileum in typical position at the bases of the crypts. Their physiological role is not yet clear. Probable functions are the production of lysozyme, proteolytic enzymes and nutritive factors for the other crypt cells, intraluminal acid-base-balance and elimination of heavy metals (OTTO, 1973). The high calcium content of these cells induced us to do further morphological studies on a probable calcium excretion by the Paneth cells in several species and under different conditions of calcium metabolism.

A systematic study of the distribution of Paneth cells in disease of the small and large intestine is described. In disease of the small intestine and appendix there is usually a fall in the number of Paneth cells normally present. Occasionally, as in Crohn's disease, there may be proliferation. In disease of the colon the number of Paneth cells is unaltered in functional and developmental disorders and increased in the inflammatory and neoplastic diseases. It is suggested that the Paneth cell proliferation originates from stem cells in the colon. The significance of an increase in Paneth cells is discussed and it is suggested that in ulcerative colitis it may produce a self-perpetuating and destructive state.

In an electron microsope study, the morphology of mature Paneth cells from the small intestine of adult rats is compared with that of differentiating Paneth cells from young rats 2 to 4 weeks old. All mature cells exhibit a marked polarity similar to that of other exocrine gland cells and contain a well developed endoplasmic reticulum, an elaborate Golgi complex, and numerous large secretory granules; they also possess an abundance of lysosomes. The most conspicuous occurrence in the process of differentiation is the development of the endoplasmic reticulum. The most immature Paneth cells possess an endoplasmic reticulum of the vesicular type, which, during maturation, is replaced by the characteristic lamellated ergastoplasm of the mature cell. At a certain stage of differentiation the cavities of the developing cisternae show numerous communications with the perinuclear space, suggesting an outgrowth of the ergastoplasm from the nuclear envelope. Furthermore, the cavities and the perinuclear space at this particular stage contain a material which shows a remarkable intrinsic periodicity. An identical periodicity was exhibited by material contained in Golgi cisternae and secretory granules. Lysosomes are also present in the differentiating cells.

Paneth cells are present in the intestinal crypts of many species, including man; an examination of the mucosal structure shows that Paneth cells accompany a morphological pattern of short crypts with concentrated mitotic and maturation activity. Absence of Paneth cells in man is accompanied by a shortening of the crypts, a lessening of crypt activity, and an abnormal surface epithelium. It is postulated that Paneth cells are specialised nutrition units that produce a secretion rich in protein and carbohydrate which acts as a favourable luminal environment for crypt cells and also the epithelial cells on the villi. Such a hypothesis is compatible with the normal distribution in man and the abnormal occurrence of Paneth cells in disease.

The origin and fate of Paneth cells were examined in duodenum, jejunum and ileum of adult female mice, using radioautography after administration of 3H-thymidine either in a single injection or in drinking water for four days or as a continuous infusion for up to ten days. The tissues were fixed by perfusion with 4% paraformaldehyde. One-micron thick, Eponembedded single or serial sections were stained with Regaud's hematoxylin, radioautographed, and counterstained with safranin O. Mitosis of Paneth cells is never observed, nor are these cells ever labeled one hour after 3H-thymidine. Hence, Paneth cells do not divide. However, a few days after single injection or prolonged administration of 3H-thymidine, labeled Paneth cells appear. The first labeled cells have tiny granules but, as the cells age, larger and larger granules are observed. Adjacent to Paneth cells are slender undifferentiated cells which show frequent mitoses and early labeling. The evidence points to some of these cells transforming into Paneth cells. Since occasionally Paneth cells degenerate, the newly-formed ones would provide replacement for those which die, thus insuring the steady state of the Paneth cell population. The renewal of this population is characterized by a turnover time of about three weeks.

The accumulation of some biogenic substances in the Paneth cell cytoplasmic granules of the mouse jejunum, ileum and colon and the effect of some drugs on this accumulation were studied, using the histochemical formaldehyde-induced fluorescence method. The biogenic substances were injected i.v. and the specific catecholamine fluorescence appeared in the cytoplasmic secretory granules of the mouse Paneth cells about 10 min after I-DOPA, dopamine, a-methyl-DOPA, noradrenaline, a-methylnoradrenaline and adrenaline administration. Nia-lamide and pargyline markedly intensified the fluorescence intensity in the Paneth cell granules after administration of the biogenic substance. Imipramine and cocaine inhibited specifically the I-DOPA and dopamine accumulation, whereas amphetamine, amitriptyline, nortriptyline, chlorpromazin, promazine and guanethidine had no effect. After nialamide pretreatment amphetamine, nortriptyline and amitriptyline were also capable of inhibiting slightly the accumulation of these biogenic substances. 5-Hydroxytryptamine or 5-hydroxytryptophan were not taken up by the Peneth cell granules even after nialamide pretreatment. The apparent mechanism of the accumulation of biogenic substances by the Paneth cell secretory granules was discussed.

9 patients are described who were found to have a deficiency of Paneth cells in the small intestinal mucosa. 7 of these had the coeliac syndrome and 2 had a more normal jejunal biopsy. A clinical picture of severe malabsorption was almost constant and, in the patients with the coeliac syndrome, there was a failure to respond to a gluten-free diet alone. Histologically the jejunal mucosa was abnormal in every case. Crypts were frequently shortened, mitotic activity was reduced, and the surface epithelium was very abnormal. Where recovery occurred, either spontaneously or after the use of steroids, the mucosa improved and Paneth cells reappeared together. The findings point to a close. relationship between Paneth cells and small intestinal epithelial integrity.

During examination of histologic sections of organs from mice which had been injected with3H-labelled tymidine every 4th hour up to 10 days, it was found that nearly 50% of the cells in the Paneth cell compartment of the intestinal crypts remained unlabelled even after 10 days of „continuous“ labelling. The number of labelled cells in the Paneth cell region after one day of labelling was not significantly different from the number after 10days of labelling.

The breakdown products resulting from the hydrolysis of chitin by lysozyme stain with Alcian Blue. A method based upon this observation has been developed for the histochemical demonstration of lysozyme activity. The application of this method to the jejunal crypts of several animal species indicates that Paneth cell granules contain lysozyme. The binding of the hydrolysis products with Alcian Blue is so strong that any other alcianophilia (e.g. of acid mucosubstances in goblet cells) can be removed selectively by washing withN-cetylpyridinium chloride and counterstaining with Basic Fuchsin and Nuclear Fast Red.

Mouse Paneth cell granules fixed with formalin have been reported to be composed of a core consisting of a polysaccharide-protein complex which is surrounded by a shell or “halo” of acid mucopolysaccharide. Electron micrographs of granules fixed with 1% OsO4 in veronal-acetate buffer show that the halo may be merely a result of shrinkage of the granule. Similar results obtained after fixation with either 1% OsO4 in s-collidine buffer containing 1% sucrose (OsO4) or with 3% glutaraldehyde in cacodylate buffer containing 1% sucrose (GAS) strengthen the belief that the halo is a shrinkage space. Fixation in GAS for 1-4 hr followed by fixation for 1 hr in OsO4, however, reduced the halo to negligible proportions. This sequence of GAS and OsO4 improved the preservation of other cytoplasmic structures also. Staining with Alcian blue failed to demonstrate acid mucopolysaccharide in the halo; therefore, the separation of granule components into a core and a halo is probably a fixation artifact.

Direct morphometric estimation of the number of intestinal crypts per 10 mm length of jejunum, the total number of cells per crypt, and the number of Paneth cells per crypt was performed using Zenker-fixed, serially sectioned small intestines from adult BALB/c mice. The mean volume of a crypt cell was estimated to be in the order of 6.6 plus or minus 0.1 times 10-4 mm-3, while the total volume of all cells in a single crypt was 1.833 plus or minus 0.02 times 10-1 mm-3. The cell population per crypt was 278 plus or minus 7 cells, of which 44 plus or minus 10 were Paneth cells. It was estimated further that 265 plus or minus 30 crypts per 10 mm length of jejunum contained 7.4 plus or minus 0.1 times 10-4 cells, including 1.2 plus or minus 0.24 times 10-3 Paneth cells. The volume of all crypt and Paneth cells per 10 mm length of jejunum was in the order of 49 plus or minus 1.2 and 8 plus or minus 1.8 mm-3, respectively. The volume of the tunica muscularis and the surface area of the crypt's lumina were estimated to be 33.5 plus or minus 2.2 mm-3 and 118 plus or minus 19 mm-2, respectively.

Human lysozyme was localized immunocytochemically at the ultrastructural level within Paneth cells of man by use of the unlabeled antibody enzyme method. Specific staining for lysozyme was observed over secretion granules in the apical cytoplasm, within the region of the Golgi apparatus and within some, but not all, lysosomes. No staining was observed within the cisternae of the rough endoplasmic reticulum or other cellular organelles. In control experiments comparing semiadjacent sections of the same Paneth cell, substitution of either normal rabbit serum for rabbit antihuman lysozyme antiserum (specificity control) or normal sheep serum for sheep antirabbit immunoglobulin G antiserum (method control) completely eliminated specific staining for lysozyme. The intensity of staining for lysozyme was related to both the titer and length of exposure to antilysozyme antiserum. Specific staining was obtained in tissue embedded in Araldite or Epon and was facilitated by etching with hydrogen peroxide. No staining was observed after prolonged fixation in glutaraldehyde or treatment with uranyl acetate in block.

Friedrich Adolf Paneth was born in Vienna on 31 August 1887. His father, Dr Joseph Paneth, was a well-known physiologist and the discoverer of certain histological cells which still bear his name. His mother Sophie ( née Schwab) was the daughter of an industrialist who was, for a time, a representative in the Austrian parliament. She had been given a classical education, which was rare at that time for a girl, and had a keen interest in the natural sciences and philosophy. Fritz had two brothers, one older and one younger, and, though both parents were of Jewish descent, the three children were brought up in the Protestant faith. He was first taught at home, but later attended Vienna’s renowned Schotten Gymnasium. He was conscientious in his school work and was also endowed throughout his life with an unusual delicacy of touch, so that it was a pleasure to watch his manipulations of familiar as well as of scientific objects. At school he was a good gymnast and later developed more than average skill as a skier, as well as becoming a keen climber. Another major interest was photography. He took up colour photography in 1908, when it was still in the experimental stage; his early achievements in this field would have been impossible without his extraordinary inborn patience. The results compare favourably in selectivity of the view and beauty of tone with many more recent efforts, and form a valuable record of one side of a full life.

The duodenal progenitor population has been investigated with tritium thymidine (H3-T) and autoradiography in male Swiss albino mice to characterize the role that individual cell types (principal, goblet, and Paneth) have in the proliferative activity of the duodenal crypt of Lieberkuhn. Incorporation of H3-T into newly synthesized DNA of all three cell types was observed by one-half after injection of the isotope. Labeled principal and goblet cells were distributed from the first cell of the cryptal mouth to the last cell at the cryptal base. The majority of labeled cells was found in the central region of the crypt Labeled Paneth cells were restricted to the last four cells at the cryptal base. Analysis of the percentage of labeled metaphase and mitotic figures after H3-T administration showed that the progenitor cycle duration of principal and goblet cells was about 15 hours, while that of Paneth cells was approximately 80 hours. The mean S-phase duration was 7.4, 7.4 and 8.7 hours for principal, goblet, and Paneth cells, respectively. Thus, the G1 portion of the cell cycle was considerably longer for Paneth cells, while all phases of the progenitor cycle were of the same duration for principal and goblet cells. It appears from the data that the three cell types of the adult duodenal crypt comprise independent populations capable of self-renewal. Our observations also suggest the existence of causal relationships between the positioning of the proliferative cells types in the duodenal microenvironment and their respective cycle durations.

The incidence of argentaffin and Paneth cells in epithelial tumours of the large intestine was investigated. Argentaffin cells were found in adenomatous polyps, villus adenomas, polyposis coli, Peutz-Jehgers9 polyps, juvenile polyposis, and adenocarcinomas. Paneth cells were not found in metaplastic or juvenile polyps. The crypt unit was destroyed in neoplasia and argentaffin and Paneth cells occurred either as a result of sequestration or were taking part in the neoplastic process. The crypt unit was retained in the disorders of epithelial growth. The identification of argentaffin and Paneth cells enabled the crypt to be defined and thus provided a useful, practical aid in the differentiation between neoplasms and disorders of epithelial growth.

Proliferation of Paneth and goblet cells of mouse duodenal crypts was studied by high resolution light microscope radioautography. In one group of mice, blood levels of thymidine-3H were sustained for up to 12 hr by repeated injections of isotope to facilitate identification of proliferating cells. In these animals, many goblet cell nuclei incorporated thymidine-3H whereas only 1 of 6261 tabulated Paneth cells was labeled. Cells intermediate in structure between undifferentiated and goblet cells and between undifferentiated and Paneth cells were identified and their light and electron microscopic features are described. A significant number of these "intermediate" cells incorporated thymidine-3H into their nuclei. Another group of mice received a single injection of thymidine-3H. These animals were killed 4 hr to 29 days after isotope administration. Goblet cells and intermediate cells with labeled nuclei were identified 4 hr after thymidine-3H but could not be seen after 15 days. In contrast, Paneth cells with labeled nuclei were not observed until 24 hr after thymidine-3H but were still present at 29 days, long after labeled undifferentiated, goblet, and intermediate cells had disappeared. We conclude that differentiated Paneth cells in mouse duodenum do not normally proliferate, but, instead, arise by differentiation from undifferentiated crypt cells or from intermediate cells. Moreover, once formed, Paneth cells persist in crypts for a prolonged period. In contrast, intermediate cells and crypt goblet cells proliferate actively and are less stable cell populations than differentiated Paneth cells. The precise function of the intermediate cells is not known, but they may represent transition forms between undifferentiated cells and the more matrure secretory cells. Damage of crypt epithelial cells, thought to be due to radiation effects, was evident in both groups of mice.

The case to be described is one of cholecystostomy for presumed mucocele in which, during a drainage period of thirty-four days, there was an excessive out-pouring of chloride and water; the source of this fluid was revealed only on subsequent cholecystectomy, when there was found in the gall-bladder a papilloma composed of intestinal epithelium. It is presented in detail on account of the difficulties in diagnosis, the peculiar biochemical behaviour, and the unique underlying pathology.

n/a

Fur die Genauigkeit der Ergebnisse radiometrischer Analysen sind neben Wage- und Pipettierfehlern vor allem die endliche Dauer der Zahlzeiten und die Unscharfe der Aktivitatseichung wesentlich. Es wird gezeigt, wie sich die Eichdaten moglichst genau und doch einfach gewinnen lassen. Der Einflus aller Fehlermoglichkeiten wird nach der Gausschen Methode der Fehlerrechnung verfolgt. Samtliche zur Berechung des „Mittleren Fehlers” eines Analysenergebnisses notigen Formeln sind angegeben; erlautert wird das Verfahren an speziellen Beispielen. Durch Kontrollanalyse wird gezeigt, das die Befunde auf wenige Prozente genau sind.

Paneth cells were found in the mucosa of the sigmoid colon in 13 out of 27 cases of diverticulosis and diverticulitis. It is concluded that their appearance at this site is a non-specific reaction to injury.