The colonic groove or furrow: A comparative morphological study of six species of African mole‐rats (Rodentia,Bathyergidae)

Herbivorous mammals such as nutrias, guinea pigs, chinchillas, and mole‐rats have a longitudinal mucosal colonic groove (furrow) in their ascending colon, which is thought to play a role in the colonic separation mechanism (CSM). It is not known whether this groove is structurally modified to adapt to this function in mole‐rat species. The morphology of this groove was studied in 32 mol‐rats, four species, one of which consisted of three subspecies, endemic to southern Africa and two species found in eastern Africa. The macroscopic morphology of the groove was documented, and samples for histological examination were taken. The groove was wide at its origin at the cecocolic junction and was lined on either side by a row of papillae with the opposing papillae slightly offset in arrangement. The papillated groove gradually decreased in size toward the distal part of the ascending colon where it disappeared. This pattern was similar in all species except in Heterocephalus glaber, where the papillae were absent and the groove was lined by two longitudinal ridges. A histological examination of cross sections revealed that the mucosa covering the inner and outer walls of the groove was rich in mucous‐secreting goblet cells. The walls of the groove contained smooth muscle extending from the inner circular smooth muscle layer at the base to the tips of the papillae in all species examined as well as arteries, lymphatic vessels, and prominent sinusoid‐like veins. The groove could be demonstrated both macroscopically and histologically in three Bathyergus suillus fetuses of varying sizes. The sinusoid‐like veins present in all grooves, regardless of macroscopic shape, suggest that they have a role in the functioning of the groove. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.     

The comparative anatomy of the abdominal gastrointestinal tract of six species of African mole‐rats (Rodentia,Bathyergidae)

The gastrointestinal tracts (GITs) of six species of African mole‐rats (Bathyergidae) were compared. The aim was to provide a comprehensive anatomical comparison between the different species. The relative shape, length, and surface areas were taken into account to determine whether the GITs are phylogenetically constrained or exhibit anatomical adaptations in response to diets. In all six species the stomach was simple and glandular. With the exception of Heterocephalus glaber, the caecum was coiled in a flat spiral, the ascending colon was arranged in a loop of varying lengths, and a mucosal colonic papillary‐lined groove was present in the ascending colon in all species. By contrast, the caecum in H. glaber was uncoiled, the ascending colon was not looped, and the groove was not papillated. A caeco‐appendix was observed only in Bathyergus suillus and Georychus capensis. Hierarchical multivariate cluster analysis on the presence/absence of nine anatomical structures associated with the GIT of mole‐rats revealed that H. glaber was anatomically the least similar of the six species (77.6% similarity) with respect to the nine GIT variables included. All Cryptomys species were the same (100% similarity), and two species B. suillus and G. capensis grouped together and were more similar to the Cryptomys genus (95% similarity) than they were to H. glaber. These findings support previous phylogenetic classifications. The voluminous caeco‐colon in B. suillus may be explained by its ingestion of grasses in addition to below‐ground storage organs of plants. We conclude that phylogeny and diet affect the GIT anatomy of the African mole rats studied here. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

Colonic epithelium reactive monoclonal antibodies

Summary We have produced a small library of colonic mucosa and colorectal carcinoma reactive monoclonal antibodies (MoAbs) by immunizations with extracts of human colon cancer tissue and a human colon cancer cell line. Hybridoma supernatants were tested on (normal and neoplastic) human tissues by immunoperoxidase methods to evaluate organ or tissue specificity. Initial biochemical characterization of the target antigens was performed by gelpermeation chromatography, Western blotting and competition assays.Based upon the immunoreactivity patterns and the characteristics of the antigen four groups of MoAbs could be distinguished. The first group concerns the antibodies PAR-LAM 3, 9 and 10. These antibodies react with an 87 kDa protein moiety in high molecular weight (2–5×10⁶ Da) glycoproteins. In intestinal and colon mucosa these antibodies showed diffuse binding with goblet cells. In colon carcinoma decreased reactivity with these MoAbs was found.The second group consists of antibodies PARLAM 8, 12 and 13. These antibodies react with large (>5×10⁶ Da) glycoproteins, most likely with carbohydrate epitopes. By immunohistochemistry in normal colon mucosa the antibodies all show granular supranuclear reactivity with goblet cells. These antibodies show increased reactivity with colon adenomas and adenocarcinomas.A third group is formed by PARLAM 2, which also reacts with a large (>5×10⁶ Da) glycoprotein, showing a granular distribution in goblet cells. In colon carcinomas more extensive expression is found than in normal colonic mucosa. Finally, the fourth group consists of PARLAM 11, which also reacts with a large (>5×10⁶ Da) glycoprotein, located in the brush border of colonic columnar cells.These antibodies might be useful tools for the analysis of the expression of mucin related glycoproteins in normal, preneoplastic and neoplastic colon mucosa.Supported by grant RL 82-1 of the Netherlands Cancer Foundation, K.W.F.     

The influence of colonizing micro-organisms on development of crypt architecture in the neonatal mouse colon

The effects of the normal colonizing microflora on postnatal development in the infant mouse were determined by comparison of crypt parameters in histological sections of the ascending colons of conventional specified-pathogen-free mice and their germ-free counterparts. Association of bacteria with the developing colonic mucosa in the third postnatal week caused a lengthening of the crypt column and depressed the total number of secreting goblet cells in each crypt. Thus the increasing bacterial burden during colonization of the developing colon was associated not only with expansion of the proliferative component of the crypt but also with modulation of the relative proportions of crypt cell populations.     

Luminal leptin activates mucin-secreting goblet cells in the large bowel

Leptin has been suggested to be involved in tissue injury and/or mucosal defence mechanisms. Here, we studied the effects of leptin on colonic mucus secretion and rat mucin 2 (rMuc2) expression. Wistar rats and ob/ob mice were used. Secretion of mucus was followed in vivo in the rat perfused colon model. Mucus secretion was quantified by ELISA, and rMuc2 mRNA levels were quantified by real-time RT PCR. The effects of leptin alone or in association with protein kinase C (PKC) and phosphatidylinositol 3-kinase (PI3K) inhibitors on mucin secreted by human mucus-secreting HT29-MTX cells were determined. Leptin was detected in the rat colonic lumen at substantial levels. Luminal perfusion of leptin stimulates mucus-secreting goblet cells in a dose-dependent manner in vivo in the rat. Leptin (10 nmol/l) increased mucus secretion by a factor of 3.5 and doubled rMuc2 mRNA levels in the colonic mucosa. There was no damage to mucosa 24 h after leptin, but the number of stained mucus cells significantly increased. Leptin-deficient ob/ob mice have abnormally dense mucus-filled goblet cells. In human colonic goblet-like HT29-MTX cells expressing leptin receptors, leptin increased mucin secretion by activating PKC- and PI3K-dependent pathways. This is the first demonstration that leptin, acting from the luminal side, controls the function of mucus-secreting goblet cells. Because the gel layer formed by mucus at the surface of the intestinal epithelium has a barrier function, our data may be relevant physiologically in defence mechanisms of the gastrointestinal tract.     

Secretory glycoconjugates of a mucin-synthesizing human colonic adenocarcinoma cell line

Summary Lectins were used to characterize mucin glycoproteins and other secretory glycoconjugates synthesized by a human colon adenocarcinoma-derived cell line which expresses a goblet cell phenotype. Despite being clonally derived, HT29-18N2 (N2) cells, like normal goblet cells in situ were heterogeneous in their glycosylation of mucin. Only wheat-germ agglutinin, which recognizes N-acetylglucosamine and sialic acid residues, and succinylated wheat-germ agglutinin, which binds N-acetylglucosamine, stained the contents of all secretory granules in all N2 goblet cells. The N-acetylgalactosamine binding lectins Dolichos biflorus and Glycine max stained 20% and 21% of N2 goblet cells respectively. Ricinus communis I, a galactose-binding lectin, stained 67% of N2 goblet cells although staining by another galactose-binding lectin, Bandeiraea simplicifolia I, was limited to 19%. Peanut agglutinin, a lectin whose Gal(1–3)GalNAc binding site is not present on mucins produced in the normal colon but which is found on most mucins of cancerous colonic epithelia, stained 68% of the cells. Ulex europeus I, a fucose-binding lectin, did not stain any N2 goblet cells. Four lectins (Lens culinaris, Pisum sativum, Phaseolus vulgaris E, Phaseolus vulgaris L) which recognize sugars normally present only in N-linked oligosaccharides stained up to 38% of N2 goblet cells. The binding of these lectins indicates either both O-linked and N-linked oligosac-charide chains are present on the mucin protein backbone or the co-existence of non-mucin N-linked glycoproteins and O-linked mucins within the goblet cell secretory granule.     

Functional morphology of the equine pelvic flexure and its role in disease. A review

The hindgut is the major site in the horse for nutrient digestion and absorption. Most of this activity occurs in the large intestinal compartments, i.e., cecum, right and left ventral colon and left and right dorsal colon. The colonic pelvic flexure is a short and narrow loop connecting the left ventral and left dorsal colon. It is not significant directly in digestive and absorptive processes but plays an important functional role in regulating colonic aboral and retropropulsive transit of digesta through its motility pacemaker activity. The pelvic flexure also contributes to the pathophysiology of colic, the leading cause of death in horses. Its narrow lumen may contribute to colonic impaction, and malfunctions of the pacemaker may contribute to volvuli and colonic displacements. Neuronal and ganglion density of the myenteric plexus is increased at the pelvic flexure and adjacent left dorsal colon pacemaker region. Contractile activity, vasoactive intestinal peptide (VIP) and neurokinins-1 and -3 are all enhanced in the pelvic flexure. The mucosa histologically resembles that of the ventral and dorsal colon, with apically-granulated principal cells and goblet cells lining the luminal surface. Clustered intranuclear inclusions resembling the cytoplasmic granules are also observed by electron microscopy in the principal cells as elsewhere in the horse colon. Further neuroendocrine and morphologic investigation of the pelvic flexure is warranted due to the great importance of this localized region for normal function and pathophysiology.     

The histochemistry of glycoconjugates in the colonic epithelium of the chicken

In the colonic epithelium of the chicken, glycoconjugates have been studied by means of selected histochemical methods of light and electron microscopy. According to the results obtained, most of the colonic goblet cells contained acidic and neutral glycoconjugates with sulphate and vicinal diol groupings, alpha-D-mannose and alpha-D-glucose residues and sialic acid-galactose dimers. These goblet cells were found to undergo changes in histochemical reactivity during upward migration along the crypts; alpha-D-mannose and alpha-D-glucose residues and terminal sialic acid-galactose dimers increased in amount. The striated border of the colonic columnar cells has, likewise, been found to contain such glycoconjugates as were similar in reactivity to those of the goblet cells. The histophysiological significances of glycoconjugates involved in the chicken colonic epithelium have been discussed with special reference to the functional activities of the carbohydrates.     

Comparative gastrointestinal morphology of Tachyoryctes splendens (Rüppell, 1835) and Heliophobius emini, (Noack, 1894) two species of East African mole‐rats

Tachyoryctes splendens (Northeast African mole‐rat) and Heliophobius emini (Emin's mole‐rat) are two African mole‐rats that represent separate allopatric rodent families namely Spalacidae and Bathyergidae respectively. While these species consume a similar diet of underground plant storage organs such as roots and tubers, T. splendens has been reported to additionally consume small amounts of aerial foliage. This study aims to provide detailed gross morphological and histological morphometric analyses of the gastrointestinal tract (GIT) of these two subterranean species. The formalin fixed gastrointestinal tracts of T. splendens (n = 9) and H. emini (n = 6) were photographed, weighed and measured. The length and basal surface areas were calculated for each anatomically distinct region. Representative histological samples were prepared and stained using Hematoxylin and Eosin. Microscopic luminal measurements were used to calculate a surface enlargement factor and the luminal surface area of each region. Tachyoryctes splendens had a large double chambered hemi‐glandular stomach with a macroscopically visible transition from keratinized stratified squamous epithelium to glandular epithelium. The cecum was large and the luminal surface revealed a single spiral fold. The ascending colon of T. splendens was arranged in a spiral, with two centripetal and two centrifugal windings. The descending colon was arranged in a single parallel loop, similar to H. emini . A narrow colonic groove was accompanied by V‐shaped folds on either side. Heliophobius emini had a simple glandular stomach, a large, haustrated cecum that displayed a cecal appendix and the descending colon was arranged in a single parallel loop. The internal aspect of the colon revealed a wide colonic groove extending from the ceco‐colic junction to distal colon. As both species originate from a similar geographical region and ingest very similar diets, it is likely that the differences in the GIT morphology are attributed to phylogeny as the species represent two different families of mole‐rats.     

Lectin histochemistry of 1,2-dimethylhydrazine-induced rat colon neoplasia

Lectins linked to fluorescein were used as carbohydrate probes to examine the goblet cell mucin and epithelial cell surface glycoconjugate alterations in an experimental rodent model of colonic neoplasia induced with parenteral 1,2-dimethylhydrazine dihydrochloride. Lectins derived from Triticum vulgare (WGA), Ricinus communis (RCA1), and Limulus polyphemus (LPA) showed reduced labeling of goblet cell mucin in these tumors, while binding with peanut lectin from Arachis hypogaea (PNA), a lectin ordinarily failing to bind to mucin in normal colon, was positive. In addition, RCA1 and LPA showed increased cell surface labeling of neoplastic epithelial cells. Finally, alterations were observed in lectin binding to "transitional" colonic mucosa adjacent to colonic tumors from carcinogen-treated rats. These findings indicate that significant alterations in both membrane and mucin glycoconjugates occur in colonic tumors and mucosa adjacent to tumors in a chemically induced experimental animal model of human colon cancer.     

Structural and biochemical differentiation of the guinea-pig colon during foetal development

Summary We have studied some aspects of the morphological and biochemical differentiation of the foetal guinea-pig colonic epithelium. At day 40 the epithelium was organised in ridges and appeared pseudo-stratified. Folding of the epithelium, followed by villus formation, occurred between days 45 and 55, and by day 50 mucus-secreting goblet cells appeared at the bases of the colonic villi. By day 55 most epithelial cells, including goblet cells, possessed numerous microvilli which, by day 65, had become organised into well developed brush-borders. Between day 55 and term (day 65–68) mucosal depth increased markedly and the colon attained its final glandular morphology.Biochemical studies showed the specific activities of the microvillar hydrolases to be much lower in the washed colon than in either foetal meconium or small intestine at all times during development. Furthermore, a membrane fraction highly enriched in microvillus hydrolase activities was prepared from foetal colonic meconium using techniques originally devised to isolate the foetal small intestinal microvillus membrane. This meconial subfraction was almost identical in polypeptide composition to the highly-purified foetal small intestinal microvillus membrane. Identification of the colonic microvillus membrane was hampered by the absence of reliable membrane markers. Nevertheless, a fraction 14-fold enriched in aminopeptidase activity was prepared from day 40 foetal colon and its polypeptide composition compared by SDS-PAGE to that of the small intestinal microvillus membrane at the same age.     

Histochemical studies of epithelial cell glycoproteins in normal rat colon

Two general classes of glycoproteins have been identified in the colonic epithelial cells of the Sprague Dawley rat. Glycoproteins belonging to the first of these classes contain sialic acids both with and without side chain o-acyl substituents, abundant o-sulphate ester and 'neutral sugars' (hexose, 6-deoxyhexose or N-acetyl hexosamine residues) with oxidisable vicinal diols and are located in the goblet cells of the descending colon and in goblet cells populating the upper halves of the crypts of the ascending colon. In the descending colon, the sulphosialoglycoproteins in the goblet cells in the base of the crypts contain sialic acids without side chain o-acyl substituents. It appears that as these cells migrate up the crypts, there is o-acylation of the side chains of the sialic acids of the glycoproteins and an increase in the quantity of 'neutral sugars' without a corresponding increase in sialic acid. Glycoproteins with similar properties to those of the goblet cells of the upper halves of the crypts of the descending colon, but containing less sulphate, are found in the goblet cells of the upper half of the crypts of the ascending colon. The second general class of glycoproteins contain sialic acids all, or almost all of which, are substituted at position C8 and only relatively small quantities of sulphate. They are located in the mucous cells of the descending colon, the deep crypt secretory cells of the ascending colon and the columnar absorptive cell brush border.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of colonizing micro-flora on the mucin histochemistry of the neonatal mouse colon

Summary Mucin histochemistry on sections of colon from germ-free and conventional mouse pups showed that all goblet cell mucins were sulphated at birth. During the first two weeks of post natal development, the pattern of mucin production in the ascending colon changed to a distribution of non-sulphated mucins towards the apical zone of the crypts and sulphated sialomucins basally. In conventional animals during the third postnatal week when the complex micro-flora of the colon was becoming established, the typical adult mucin distribution pattern developed, with sulphated mucins now confined to the upper third of the crypt. However, in the absence of a colonizing micro-flora crypt mucins become more and more sulphated until at weaning, most goblet cells of the ascending colon were producing fully or partially sulphated mucins, except for one or two cells at the very base of the crypt.     

Human Intestinal Circadian Clock: Expression of Clock Genes in Colonocytes Lining the Crypt

Biological clock components have been detected in many epithelial tissues of the digestive tract of mammals (oral mucosa, pancreas, and liver), suggesting the existence of peripheral circadian clocks that may be entrainable by food. Our aim was to investigate the expression of main peripheral clock genes in colonocytes of healthy humans and in human colon carcinoma cell lines. The presence of clock components was investigated in single intact colonic crypts isolated by chelation from the biopsies of 25 patients (free of any sign of colonic lesions) undergoing routine colonoscopy and in cell lines of human colon carcinoma (Caco2 and HT29 clone 19A). Per‐1, per‐2, and clock mRNA were detected by real‐time RT‐PCR. The three‐dimensional distributions of PER‐1, PER‐2, CLOCK, and BMAL1 proteins were recorded along colonic crypts by immunofluorescent confocal imaging. We demonstrate the presence of per‐1, per‐2, and clock mRNA in samples prepared from colonic crypts of 5 patients and in all cell lines. We also demonstrate the presence of two circadian clock proteins, PER‐1 and CLOCK, in human colonocytes on crypts isolated from 20 patients (15 patients for PER‐1 and 6 for CLOCK) and in colon carcinoma cells. Establishing the presence of clock proteins in human colonic crypts is the first step toward the study of the regulation of the intestinal circadian clock by nutrients and feeding rhythms.     

Chemical and histochemical studies of normal and diseased human gastrointestinal tract. I. A comparison between histologically normal colon, colonic tumours, ulcerative colitis and diverticular disease of the colon

Summary Chemical and histochemical methods were used to compare the epithelial glycoproteins from formalin-fixed surgical specimens of normal human large intestine, colonic tumours, ulcerative colitis and diverticular disease. All the epithelial glycoproteins contained fucose, galactose, glucosamine, galactosamine and, in addition, sialic acids both with and withoutO-acyl substituents in the side chain and/or at position C₄. The glycoproteins of the normal ascending and descending colons differed significantly with respect to the percentage of the sialic acids released following digestion of the de-O-acylated glycoprotein withVibrio cholera neuraminidase and to the molar fucose-sialic acid ratio. Statistical analysis of the chemical data showed that (a) compared to normal, the sialic acids of the tumour and ulcerative colitis glycoproteins from the descending colon were significantly less substituted in the side chain and at position C₄; (b) theO-acetyl substitution pattern of the sialic acids of the ulcerative colitis glycoproteins from the ascending colon and the quantitative composition of the carbohydrate prosthetic groups of the ulcerative colitis glycoproteins from both ascending and descending colons differed from normal; (c) it was not always possible to distinguish between the ulcerative colitis and tumour glycoproteins on the basis of theO-acetyl substitution pattern of their sialic acids; and (d), there were minor differences between normal glycoproteins and those from cases of diverticular disease.     

c-kit immunoreactive interstitial cells of Cajal in the human small and large intestine

 c-kit immunohistochemistry was performed on unfixed frozen sections of human small (duodenum, jejunum, and ileum) and large intestine (ascending, transverse, descending, and sigmoid colon). The c-kit immunoreactive cells in the muscularis externa of the intestinal wall were identified as interstitial cells of Cajal (ICC) and mast cells. ICC were identified by their morphology, localization, and organization based on previous light and electron microscopic studies. In the small intestine, ICC were located primarily in relation to the myenteric plexus of Auerbach, but also in septa between circular muscle lamellae. In the large intestine, ICC were seen in relation to Auerbach’s plexus, but also and in great numbers in the circular muscle layer and in teniae of the longitudinal muscle layer. The morphology of the ICC was similar in the small and large intestine, but the pattern of distribution was obviously different. c-kit immunoreactive mast cells were found predominantly in the inner part of the circular muscle layer. The anti-c-kit method is found to be an easy and reliable method to study at least most of the interstitial cells of Cajal and thereby contribute to further normal and pathological studies. Accepted: 31 July 1997     

Morphological and histochemical characterisation of the mucosa of the digestive tract in matrinxã Brycon amazonicus (Teleostei: Characiformes)

This study describes anatomical, histological and histochemical features of the digestive tract mucosal layer of the matrinxã Brycon amazonicus, an omnivorous freshwater fish endemic from the Amazon basin. This species presents short thick oesophagus with longitudinal folds, that allow the passage of large food items. The mucosa is lined with a stratified secretory epithelium rich in goblet cells that secrete neutral and acid mucins. The two mucin types provide different viscosity in anterior and posterior oesophagus related to the protective and lubricant functions, respectively. The stomach is a highly distensible Y-shaped saccular organ. Here, it is proposed that this anatomical shape plays an essential role in food storage when food availability is abundant. The stomach mucosa is composed of epithelial cells with intense neutral mucin secretion to protects against gastric juice. The intestine is slightly coiled and presents internally a complex pattern of transversal folds that increases the absorption surface and the retention time of food. Goblet cells in the intestine secrete acid and neutral mucins that lubricate the epithelium and aid in the digestive processes. In the rectum, an increase in goblet cells population occurs that may be related to better lubrication.     

Some morphological and histochemical studies on the intestinal tract of the Brazilian sloth (Bradypus tridactylus)

The intestinal of the 3-toed sloth, Bradypus tridactylus, was studied macroscopically, with light microscope and with histochemical methods for mucosubstances. Macroscopically, the inner surface of the duodenum shows longitudinal and circular folds. There is no caecum, nor appendix. The large intestine consists of a short colon and a large rectal pouch, which has a thick wall. The mucosa of the small intestine has long leaf-shaped villi covered with columnar epithelium having a well developed striated border, and the goblet cells are scattered among the columnar cells. An association between neutral and acidic mucosubstances was detected in the goblet cells. The duodenal (Brunner's) glands are confined exclusively in the lamina propria of the duodenum. No Paneth cells were observed in the crypt lining. Argyrophil and argentaffin cells were found in the entire length of the intestine. The large intestine does not possess villi, but many goblet cells were observed in its mucosa.     

OGT Controls the Expression and the Glycosylation of E‐cadherin,and Affects Glycosphingolipid Structures in Human Colon Cell Lines

Colorectal cancer (CRC) affects both women and men living in societies with a high sedentary lifestyle. Amongst the phenotypic changes exhibited by tumor cells, a wide range of glycosylation has been reported for colon cancer‐derived cell lines and CRC tissues. These aberrant modifications affect different aspects of glycosylation, including an increase in core fucosylation and GlcNAc branching on N‐glycans, alteration of O‐glycans, upregulated sialylation, and O‐GlcNAcylation. Although O‐GlcNAcylation and complex glycosylations differ in many aspects, sparse evidences report on the interference of O‐GlcNAcylation with complex glycosylation. Nevertheless, this relationship is still a matter of debate. Combining different approaches on three human colon cell lines (HT29, HCT116 and CCD841CoN), it is herein reported that silencing O‐GlcNAc transferase (OGT, the sole enzyme driving O‐GlcNAcylation), only slightly affects overall N‐ and O‐glycosylation patterns. Interestingly, silencing of OGT in HT29 cells upregulates E‐cadherin (a major actor of epithelial‐to‐mesenchymal transition) and changes its glycosylation. On the other hand, OGT silencing perturbs biosynthesis of glycosphingolipids resulting in a decrease in gangliosides and an increase in globosides. Together, these results provide novel insights regarding the selective regulation of complex glycosylations by O‐GlcNAcylation in colon cancer cells.