Endocrine cells of mucosal epithelium in the distal part of the intestine of Lacerta vivipara

The epithelium of the distal part of intestine of the lizard Lacerta vivipara has been studied by light and electron microscopy. The total number of endocrinocytes (argyrophilic cells) was found to increase from small bowel (57 +/- cell/mm2) to colon (9 +/- 69), and cloaca (99 +/- 8). Although the number of argentaffin cells increases from the small bowel to colon, cell decrease occurs from colon (42 +/- 6 cell/mm2) to cloaca (65 +/- 10 cell/mm2). On electronograms of the colon mucosal epithelium four types of endocrinocytes were identified. Type I--with secretory granules polymorphic for the size and form, with a high electron density core, and average size 206 +/- 31 nm. Type II--with secretory granules 265 +/- 20 nm in size, having spherical form and highly electronic dense contents. Type III--contains largest (350 +/- 12 nm), spherical, oval or irregularly-shaped secretory granules, with contents of various electronic density. Type IV--endocrine cells having small (176 +/- 5 nm) spherical or oval secretory granules with a highly electronic dense core. Besides, "mixed" cells were identified, whose cytoplasm contained simultaneously mucous and endocrinous granules.     

Endocrine cells in the rabbit colon mucosal epithelium

The epithelium of the rabbit colon was studied by light and electron microscopy. The highest number of endocrinocytes in colon are observed in terminal parts of colon, i.e. in a distal part of appendix (135 +/- 15 cells/mm2) and in rectum (142 +/- 20), to decrease in the ileocaeal region (caecum proxinmal part--39 +/- 9, colon proximal part--56 +/- 9), where the least number of cells was marked. Agrentaffin cells (EC) number the same way, however, with a weaker difference in the number of cells between terminal departments and ileocaeal region. An electron microscope study of mucosal epithelium of the colon enabled us to identify 5 types of endocrinocytes. I-III types: EC-, D- and L-cells. IV and V are seldom met types, the same way as the "mixed" cells have been indentified. Whose cytoplasm simultaneously contained both mucous and endocrine granules. The received data show a certain degree of similarity in the endocrine apparatus of the rabbit with that of humans, although essential differences exists in regards of the appendix pattern.     

Quantitative characteristics of the endocrine cells of the duodenal glands of Carnivora

In the duodenal glands of the Carnivora investigated endocrine elements have been revealed, a part of them is presented as serotonin-producing EC-cells. Endocrine cells are situated in terminal parts and in glandular ducts, among them elements of open and close types are distinguished. Distribution of these cells in the glandular lobules is subjected to the distal gradient regularity, specific for the gastrointestinal tract mucosal membrane. Amount of endocrinocytes in the glands is much less than in the gut crypts. There is no correlation between distribution of the endocrine cells in the glands and in the crypts. The results of unifactor analysis of variance demonstrate a slight effect of the taxonomic position of the species on the number of endocrine cells in the duodenal glands. The proper endocrine apparatus of the duodenal glands is supposed to produce a local regulatory influence on the secretory activity of exogenic glandulocytes, as well as ensure humoral connections of the duodenal glands with other parts of the gastrointestinal tract.     

Structure and cytophysiology of gastric endocrine gland cells after repeated administration of testosterone propionate

By means of light, electron microscopy and morphometry methods contents and structural organization of endocrinocytes have been investigated in gastric glands of rats, to whom testosterone propionate has been injected for 5, 10, 15, 20 days. The hormone produces certain stimulating effect on the endocrine apparatus function. During the first stages of the experiment most of the endocrinocytes are at the state of active secretion; this is testified as a decreasing composition of histochemically revealed cells in epithelium and their degranulation. During the following stages the process of synthesis is activated in the cells and their amount increases. Appearance of numerous agranular, poorly differentiated endocrinocytes and exo-endocrine cells demonstrates certain intensification of cytogenesis of endocrinocytes in the gastric epithelium. The changes observed are considered as adaptive, directed to stabilize homeostasis, when the hormonal balance is disturbed.     

Differentiation of endocrinocytes in the small-intestinal epithelium of albino rats in ontogenesis]

By means of the light microscopy method differentiation and dynamics on contents of endocrinocytes have been studied in epithelium of the initial part of the jejunum, of the caudal part of the ileum and of the middle part between them during pre- and postnatal periods of development. Formation of the endocrine apparatus of the mucous membrane epithelium in the small intestine takes place simultaneously with tissue differentiation of the epithelial layer. The population density of endotheliocytes of the mucous membrane is maximal in the initial part of the jejunum and decreases gradually in the caudal direction. By the 15th day of the postnatal development endocrinocytes in composition of the epithelium in the small intestine reach a high level of the specific differentiation and their amount increases by 1.3 times in comparison with that in newborn animals.     

Histotopography of colon endocrine cells in the mucosal epithelium of some mammals and in man

Histotopography of colon endocrine cells in the mucosal epithelium of adult rabbits, white rats and man was studied by light microscopy. The number of endocrinocytes in rabbit and human colon was seen to increase towards the rectum, which reflects a general tendency in mammals. In the appendices of the investigated subjects, essential distinctions in endocrine cell contents were revealed. So, in rabbits, the maximum quantity of endocrine cells (135 +/- 15 cell/mm2) was observed in the appendix if compared to other parts of the colon (except rectum), whereas in the human appendix, the number of endocrine cells is minimal (13 +/- 3 cell/mm2). In all parts of rabbit and rat caeca endocrine cell contents are similar to those in the nearby part of the colon, which suggested that according to the given parameter the caecum shows no specifity.     

Seasonal dynamics of the gastric APUD cell count in a hibernator

Light microscopy was applied to study variation in the composition and number of apudocytes in the gastric epithelium of red-cheeked sousliks over time. During hibernation, the content of serotonin-producing EC cells was elevated. This is in agreement with serotonin involvement in the maintenance of the hibernation state. The number of non-EC cells taking part in digestion, which does not occur during hibernation, was lowered. Spring awakening was accompanied with an increase in the number of non-EC cells and the lack of significant changes in cells. In souslics, which were active in winter, the content of non-cells was high, since the animals continued feeding on, and the content of EC-cells was elevated similarly to that in hibernating animals, which could be explained by genetic dependence. The number of apudocytes in the gastric mucosa was discovered to correlate in the main with the features of their function at different seasons.     

Expression of ENaC and serum- and glucocorticoid-induced kinase 1 in the rat intestinal epithelium

Increase in epithelium sodium channel (ENaC) activity induced by aldosterone in the distal tubule of the kidney has been attributed to serum- and glucocorticoid-induced kinase 1 (sgk1). The distal colon constitutes another classical aldosterone-responsive epithelium that expresses both ENaC and sgk1 in an aldosterone-dependent manner. However, the site of expression and the temporal relationship of the aldosterone induction of these two proteins have not been investigated. Here, we examined the distribution and abundance of sgk1 in the rat intestine under basal conditions and after changes in the concentration of aldosterone and glucocorticoids. Results indicate that sgk1 is expressed in the distal colon and also in the ileum and jejunum. Abundance of sgk1 was high in control animals, and it did not change significantly after sodium depletion or after a single dose of aldosterone; however, it decreased after adrenalectomy. In contrast, the three subunits of ENaC were markedly induced in the distal colon by acute and chronic increases in aldosterone levels. Results indicate differential regulation of sgk and ENaC subunits by aldosterone in the distal colon. Distribution of sgk1 in the intestine beyond the aldosterone-responsive segments suggests that sgk1 may additionally regulate other sodium transporters in the intestinal epithelium.     

Changes in number of serotonin-containing cells and serotonin levels in the intestinal mucosa of rats with colitis induced by dextran sodium sulfate

The role of serotonin in the pathogenesis of inflammatory bowel disease has not been fully studied. We examined the changes in the serotonin level and the density of serotonin-containing enterochromaffin (EC) and mast cells in the intestinal mucosa of dextran sodium sulfate (DSS)-induced colitis in rats. Rats were treated with 1.5% DSS for 1 month. Serotonin levels were biochemically measured and the density of epithelial EC cells and mucosal mast cells was quantified by serotonin immunohistochemistry. DSS caused malnutrition due to chronic diarrhea. Infiltrated inflammatory cells were microscopically observed in the colonic wall with intact epithelium. The serotonin content in the mucosa/submucosa tissue was increased in the proximal and distal colon in DSS-treated rats, compared to that in control rats. The density of EC cells in the epithelium also increased in the proximal and distal colon in DSS-treated rats. In contrast, the density of mast cells in the lamina propria dramatically increased in the distal, but not in the proximal colon in DSS-treated rats. This discrepancy implies the serotonin released from EC cells and from mast cells may play different roles in the pathogenesis of DSS-induced colitis. Accepted: 30 July 1999     

Various aspects of organogenesis of the colon in Gallus domesticus: morphologic observation

The authors have investigated the morphological aspects of the wall components of the developing colon in the chick embryo (Gallus domesticus) from the 7th to th 15th day of incubation. Particular attention has been given to the lumen recanalization, phenomenon which occurs also in other animal species. The most significant results can be summarized as follows: 1) the lumen is recanalized at the 7th day only at the proximal part of the colon (Fig. 1, Tav. 1), while at the distal tract it is still completely filled by an epithelial plug (Fig. 2, Tav. 1). Therefore the recanalization of the lumen takes place cranio-caudad. 2) At the 8th day the process of recanalization of the lumen shows, in the distal part of the colon, well defined modalities. Radially oriented intraepithelial spaces within the epithelium filling the lumen join other semilunar intercellular spaces, which are placed near the central part of the occluded lumen (Fig. 3). By the junction of a couple of radially oriented spaces with one semilunar space, an U-shaped intercellular space derives, which delimits an incoming epithelial fold (Tav. 3). Such a phenomenon is continued also during the 9th and 10th day of incubation (Fig. 6, Tav. 2). 3) At the 11th day the colonic lumen is completely open and, in its distal part, the appearance of the primordial previllous ridges can be observed (Fig. 7). In the proximal tract the previllous ridges develop one day later (Fig. 8). 4) At the 13th day, in the distal part of the colon, the first appearance of crypts occurs (Fig. 10). So, while the process of recanalization of the lumen is cranio-caudad, the formation of previllous ridges and crypts proceeds caudo-cranially. 5) From the 11th day onwards the lamina propria is actively involved in the process of formation of the previllous ridges. Only at the 14th day, in the distal part of the colon anlage, the appearance of the muscularis mucosae is observed (Fig. 11). 6) The muscle layer and the subserous stratum do not show appreciable morphological changes in the course the considered period of incubation.     

Fast renewal of the distal colonic mucus layers by the surface goblet cells as measured by in vivo labeling of mucin glycoproteins

The enormous bacterial load and mechanical forces in colon create a special requirement for protection of the epithelium. In the distal colon, this problem is largely solved by separation of the bacteria from the epithelium by a firmly attached inner mucus layer. In addition, an outer mucus layer entraps bacteria to be cleared by distal transport. The mucus layers contain a network of Muc2 mucins as the main structural component. Here, the renewal rate of the inner protective mucus layer was studied as well as the production and secretion of Muc2 mucin in the distal colon. This was performed by intraperitoneal injection of N-azidoacetyl-galactosamine (GalNAz) that was in vivo incorporated during biosynthesis of O-glycosylated glycoproteins. The only gel-forming mucin produced in the colon is the Muc2 mucin and as it carries numerous O-glycans, the granulae of the goblet cells producing Muc2 mucin were intensely stained. The GalNAz-labeled glycoproteins were first observed in the Golgi apparatus of most cells. Goblet cells in the luminal surface epithelium had the fastest biosynthesis of Muc2 and secreted material already three hours after labeling. This secreted GalNAz-labeled Muc2 mucin formed the inner mucus layer. The goblet cells along the crypt epithelium accumulated labeled mucin vesicles for a longer period and secretion of labeled Muc2 mucin was first observed after 6 to 8 h. This study reveals a fast turnover (1 h) of the inner mucus layer in the distal colon mediated by goblet cells of the luminal surface epithelium.     

Immunohistochemical study on the morphology of enterochromaffin cells in the human fundic mucosa

Summary The morphology of enterochromaffin (EC) cells in the human fundic mucosa was investigated at the lightmicroscopic level by means of the unlabeled peroxidase anti-peroxidase method, with the use of a highly specific anti-serotonin serum.EC-cells in the human fundic mucosa were sparsely distributed below the neck portion of the gland, but were found to be rather numerous in its lower half. Immunohistochemistry revealed marked pleomorphic and seemingly polynuclear EC-cells or cells with long, sometimes multipolar cytoplasmic processes. In addition, luminal contacts and contiguity between EC-cells, or interglandular connections were also encountered.The present immunohistochemical procedure permits, for the first time, a clear-cut morphological visualization of the entire population of EC-cells, and reveals the distinctive morphological features of these cells in the human fundic mucosa. These morphological findings imply that EC-cells in the fundic mucosa may be crucial in gastric function.     

"Mixed" endocrine gland cells of the duodenal epithelium in various vertebrate animals and man

Epithelium of the duodenal mucous membrane has been studied in frog, rat and man by means of electron microscopy. Together with typical endocrinocytes, incretory cells of "mixed" type are revealed in the epithelium. They are classified into two types: 1. exo-endocrinic containing in their cytoplasm simultaneously: a) mucous and EC-granules, b) mucous and S-granules, c) granules specific for Paneth's cells and S-granules, d) granules specific for Paneth's cells and L-granules; 2. endocrinocytes with incretory granules of various types: with D and I, with D and EC, with D and L granules. Various ultramicroscopic organization of the "mixed" exo-endocrinic cells is demonstrated. It can be considered as various stages of their differentiation. In cytoplasm of the "mixed" cells of the second type (endocrinic) a constant presence of D-granules is revealed, that is, evidently, characteristic, within the limits of the gastro-entero-pancreatic system, for the cells with two types of the endocrinic granules.     

Anion transport in the colon

The principal anions transported by colonic epithelium are Cl-, HCO3- and organic anions (OA-), particularly acetate, butyrate and pyruvate, these last being formed by microbial degradation of carbohydrate. In the normal absorptive rat colon, Cl- is transported from lumen to plasma both by the transcellular and paracellular pathways. The transcellular route appears to depend on amiloride-insensitive coupling of Na+-Cl- at the mucosal (apical) membrane, the Na+ electrochemical gradient energizing Cl- uptake. Intraluminal [HCO3-] rises as Cl- as absorbed, and a mucosal Cl- -HCO3- exchange carrier has been postulated. In some species (and in distal colon of the rat when sodium-depleted), the putative Na+-Cl- carrier is absent so that Cl- absorption then depends largely on the paracellular electrochemical gradient. Absorption of OA- is independent of the transepithelial p.d., is associated with HCO3- secretion and is considerably reduced by acetazolamide. In the absence of Cl-, OA- supports Na+ absorption but does not depend on it continuing unchanged when the latter is blocked. Colonic epithelium can become secretory and an example of this state is congenital chloridorrhoea in which an elevated transepithelial p.d. is associated with excessive Cl- secretion. Here, it appears that the Na+-Cl- and Cl- -HCO3- carriers are lost and Cl- conductance of the mucosal membrane substantially increased. The transepithelial uphill movements of Cl- or HCO3- in the absorptive and secretory colon appear to depend on coupling to other ionic flows, and there seems to be no need to postulate active transport of these ions.     

Studien über die endokrinen Zellen des Magendarmtraktes

Zusammenfassung Die serotoninhaltigen, nach Formaldehydbedampfung im UV-Licht gelbfluoreszierenden EC-Zellen im Magendarmepithel wurden bei normalen und bei tryptophanfrei ernährten Ratten untersucht. Bei Kontrolltieren ist die Zahl der EC-Zellen, bezogen auf die Schleimhautfläche, im Duodenum größer als im Pylorus. Im Duodenum sind die EC-Zellen etwa gleichmäßig auf Oberflächen- und Drüsenepithel verteilt, im Pylorus ganz überwiegend im Drüsenepithel lokalisiert. Die Brunnerschen Drüsen besitzen keine EC-Zellen. Tryptophanfreie Diät führt zu starker Abnahme des Serotoningehalts der EC-Zellen, nach Umsetzung auf Normalkost steigt er wieder an. — Die Ergebnisse sprechen weiterhin dafür, daß die EC-Zelle sowohl Speicher- als auch Syntheseort von Serotonin ist. Dabei kommt Serotonin in der EC-Zelle wahrscheinlich in zwei Fraktionen vor, nämlich zytoplasmatisch gelöst und granulagebunden. Die histochemischen Nachweismethoden für Serotonin, Möglichkeiten des Eingriffs in die verschiedenen Syntheseschritte des Serotonins und deren Spezifität in bezug auf die EC-Zelle als physiologischem Ort der Serotoninsynthese werden diskutiert.

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Studies on the endocrine cells of the gastrointestinal tractII. Fluorescence microscopy of the EC-cells after tryptophan-free diet

Summary The serotonin-containing EC-cells of the gastro-intestinal mucosa, which show yellow fluorescence after treatment with gaseous formaldehyde, were investigated in the rat after normal and tryptophan-free nourishment. In control animals the number of EC-cells, related to epithelium area is higher in the duodenum than in the pyloric region. In the duodenum the EC-cells show about the same distribution in surface epithelium and gland epithelium. In the pyloric region EC-cells are localized predominantly in the gland epithelium. No EC-cells were found in the Brunner glands. After tryptophan-free diet the serotonin-level of the EC-cells strongly decreases; serotonin-level increases after return to normal nourishment. The results also suggest, that serotonin is synthesized as well as stored in the EC-cell and that it occurs in two fractions: bound to granules and dissolved in the cytoplasm. Discussion deals with the histochemical methods for determination of serotonin, the possibilities of influencing the pathways of serotonin metabolism and the specificity of these possibilities concerning the EC-cell as the physiological site of serotonin synthesis.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft, Antrag Fo 77, 1–4.     

Enterochromaffin (EC-) cells of the mammalian gastro-entero-pancreatic (GEP) endocrine system: cellular source of pro-dynorphin-derived peptides

Summary It has long been disputed whether mammalian enterochromaffin (EC-) cells contain a peptide in addition to serotonin. Previous immunohistochemical studies have provided evidence for the presence of enkephalins in EC-cells. These findings, however, are equivocal. Therefore, the problem of opioid peptides in EC-cells has been re-examined in the gastro-intestinal mucosa of dog, guinea-pig and man. A battery of antisera against derivatives of pro-opiomelanocortin, pro-enkephalin and pro-dynorphin have been applied to semithin serial sections of the tissues, in combination with fluorescence histochemistry and serotonin immunocytochemistry. Our findings indicate that EC-cells of the investigated species contain pro-dynorphin-related peptides, i.e. dynorphin A and -neo-endorphin, but no derivatives from pro-opiomelanocortin or pro-enkephalin. Since remarkable interspecies variations occur with respect to the number and staining characteristics of opioid immunoreactive EC-cells, it is concluded that pro-dynorphin shows specific routes of post-translational processing depending upon the species and the gastro-intestinal segment investigated. Future studies should focus on the mutual relationships between serotonin and dynorphins and on the physiological significance of these peptides in the gastrointestinal tract.Part of the results were presented at the Bayliss and Starling Society National Scientific Meeting 1985, London (Cetin et al. 1985)     

Luminal mucin in the large intestine of mice,rats and guinea pigs

Summary The luminal and epithelial mucin was studied histochemically in the large intestine of mice (Mus musculus), rats (Rattus rattus) and guinea pigs (Cavia porcellus) using freeze-substitution and vapor-fixation methods. Neutral mucin decreased and acid mucin increased in the epithelium from the cecum to the distal colon. Vacuolated cells contained more acid mucin than goblet cells. Luminal mucin always contained neutral mucin, which formed the main constituents in the cecum and in the proximal colon. Sialo-mucin increased from the cecum to the distal colon. Sulfo-mucin appeared only in the distal colon. Except in the cecum a luminal mucin layer (LML) was found at the epithelial surface. In the proximal colon LML was not entirely continuous and varied in composition and thickness (182.4 ± 170.1, 150.5 ± 110.4, 30.0 ± 28.9 (m), in mice, rats and guinea pigs, respectively), and contained many bacteria. In the distal colon LML was compact, homogeneous and thin (33.6 ± 18.8, 16.1 ± 7.3, 29.1 ± 20.0 (m), in mice, rats and guinea pigs, respectively) containing few bacteria. Possible functions of the luminal mucin and their regional differentiations were discussed.Supported by a grant from Deutsche Forschungsgemeinschaft (En 65/9). A preliminary part of this study was presented at 5th ISRP (September 1979), Clermond-Ferrand, France. Authors thank Miss G. Becker for her technical assistance     

The thickness of the mucus layer in different segments of the rat intestine

Summary The thickness of the pre-epithelial mucus layer has been measured in different gut segments of rats kept under normal (ad libitum) feeding conditions, and after 48 h of fasting, using cryostat sections and celloidin stabilization from samples containing luminal contents. The mucus layer of the stomach, duodenum, jejunum, ileum, caecum, proximal colon, colon transversum, distal colon and rectum was studied in five groups of male rats (10, 40, 70 and 150 days of age, and older). Underad libitum feeding conditions, a distinct and continuous mucus layer, with a thickness of more than 3 μm, was only observed in the colon transversum, in the distal colon, in the rectum and in the stomach. No pre-epithelial mucus layer was observed in the duodenum and jejunum where the glycocalix from the apical membrane of the superficial cells appeared to be in a direct contact with the luminal ingesta. In the ileum, caecum and the proximal colon, the surface epithelium of the mucosa was only partly covered by a mucus layer of highly variable thickness. After 48 h of fasting, a mucus layer of 28.8 ± 25.6 μm and 93.3 ± 59.4 μm thickness, respectively, was found in the duodenum and jejunum of adult rats, but no increase in the thickness of the mucus layer was observed in the rat hind gut.     

Control of ion transport in mouse proximal and distal colon by prolactin.

The lactogenic hormone prolactin (PRL) has been known to affect Ca(2+) and electrolyte transport in the intestinal epithelium. In the present study we analyzed ion transport in mouse proximal and distal colon, and acute changes induced by PRL. In the proximal colon, carbachol activated a Ca(2+) dependent Cl(-) secretion that was sensitive to DIDS and NFA. In the distal colon, both ATP and carbachol activated K(+) secretion. Ca(2+) -activated KCl transport in proximal and distal colon was inhibited by PRL (200 ng/ml), while amiloride sensitive Na(+) absorption and cAMP induced Cl(-) secretion remained unaffected. Luminal large conductance Ca(2+) -activated K(+) (BK) channels were largely responsible for Ca(2+) -activated K(+) secretion in the distal colon, and basolateral BK channels supported Ca(2+) -activated Cl(-) secretion in the proximal colon. Ca(2+) chelating by BAPTA-AM attenuated effects of carbachol and abolished effects of PRL. Both inhibition of PI3 kinase with wortmannin and blockage of MAP kinases with SB 203580 or U 0126, interfered with the acute inhibitory effect of PRL on ion transport, while blocking of Jak/Stat kinases with AG 490 was without effects. PRL attenuated the increase in intracellular Ca(2+) that was caused by stimulation of isolated colonic crypts with carbachol. Thus PRL inhibits Ca(2+) dependent Cl(-) and K(+) secretion by interfering with intracellular Ca(2+) signaling and probably by activating PI3 kinase and MAP kinase pathways.