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The colon is believed to absorb NaCl via the coupled operation of apical Na/H exchanger-3 (NHE3) and Cl/HCO(3) exchanger SLC26A3 (DRA). Efficient coupling requires that NHE3 and DRA operate in close proximity within common luminal and cytosolic microenvironments. Thus we examined whether these proteins coexist along the apical margin of surface enterocytes by quantitative immunofluorescence microscopy in consecutive colon segments from nonfasted mice and rats. The cecocolonic profiles of NHE3 and DRA expression were roughly inverse; NHE3 was highest in proximal colon (PC) and negligible in distal colon, whereas DRA was absent in early PC and highest in the late midcolon, and DRA was prominent in the cecum whereas NHE3 was not. NHE3 and DRA coexisted only in the middle third of the colon. The consequences of unpaired NHE3/DRA expression on mucosal surface (subscript MS) pH and Na(+) concentration ([Na(+)]) were assessed in nonfasted rats in situ using miniature electrodes. In the cecum, where only DRA is expressed, pH(MS) was approximately 7.5, markedly higher than underlaying stool (6.3), consistent with net HCO(3)(-) secretion. In the early PC, where NHE3 is not expressed with DRA, pH(MS) was acidic (6.2), consistent with unopposed H(+) secretion. [Na(+)](MS) was approximately 60 mM in the cecum, decreased along the PC to approximately 20 mM, and declined further to approximately 10 mM distally. Cl(-) was secreted into the PC, then reabsorbed distally. Our results suggest a model in which 1) unpaired DRA activity in the cecum maintains an alkaline mucosal surface that could neutralize fermentative H(+); 2) unpaired NHE3 activity in the early PC preserves an acidic mucosal surface that could energize short-chain fatty acid absorption; and 3) coupled NHE3/DRA activities in the midcolon allow for vigorous NaCl absorption at a neutral pH(MS).

In the mammalian cecum and colon, a single layer of absorptive, mature enterocytes are a crucial element of the physical barrier to the contents of the lumen. Enterocytic differentiation involves expansion of cytoplasmic cytoskeletal networks, which have been proposed to maintain structural integrity of individual cells and thus the entire epithelial barrier. We sought molecular tools to test this hypothesis in vivo, because in vitro systems displaying full intestinal epithelial differentiation have not yet been developed. Vav proteins are RhoGEFs that modulate cytoskeletal networks in immune cells. We found that Vav proteins were preferentially expressed in terminally differentiating cecal and colonic enterocytes. Loss of Vav protein expression in triple-knockout mice (Vav1(-/-);Vav2(-/-);Vav3(-/-)) resulted in defective expansion of microtubule cytoskeletons, a significant decrease in cell height and diminished expression of differentiation markers. Despite these changes, enterocytes in the triple-mutant mice did not contain measurable alterations in actin cytoskeleton, apical cell-cell junctions, nuclear position or global polarized delivery of proteins involved in terminal differentiation. Aged triple-mutant mice spontaneously developed ulcerative lesions that were, in part, a result of defective wound repair. These studies show that Vav proteins are required for enterocytic differentiation and colonic epithelial barrier integrity.

The purpose of this study was to establish and validate fully automatic measurement of cell proliferation on scanned images of rat cecum and colon. Tissue slides were taken from a 4-week mechanistic study and processed for BrdU immunohistochemistry. Four sections of the cecum and colon per slide were scanned with the Zeiss MIRAX SCAN and transferred to the Definiens eCognition Analyst LS5.0 system for evaluation. Two rule sets for automatic counting of BrdU-positive and negative nuclei from mucosal cells on the image tiles were created by Definiens, one for cecum, one for colon. For validation, manual counting of 16 randomly selected tiles from five different slides of colon and cecum was performed. Negative and positive cell nuclei were counted in each image tile by four different people. Comparison of results from manual counting with the automatic counting showed that the sum as well as single tile data and labeling index (LI) from automatic counting were within the range of manual counting results +/-10%. Automatic counting included only cell nuclei within the mucosa whereas muscularis and lymphoid tissue as well as wrinkles from tissue preparation were excluded. In addition, two data sets from automatic counting of the same image tile were compared: (1) data where image tiles with incorrect detection of mucosa were excluded from further calculation of LI and area, and (2) data where no visual check was performed and all measurements were included. Results were very similar for both data sets. The necessity of the manual correction may therefore be doubted.

Colonic cell proliferation is the prerequisite for the genesis of cancer. Experimental and epidemiologic evidence indicate dietary factors to be one of the commonest predisposing factors in the development of several types of cancers including large intestine. Here we have investigated the role of the fruit pulp of Tamarindus indica L. (TI), a tropical plant-derived food material, on the proliferating colonic mucosa using Swiss albino mice. Crypt cell proliferation rate (CCPR), on histological basis and [3H]-thymidine incorporation assay were chosen to evaluate the modulating potential of TI per se and in response to a subacute dose of N-nitroso N'-methyl urea (MNU). Descending colonic segment showed greater rate of cell proliferation than the ascending colon and cecum tissues isolated from the group 2 (TI-per se) when compared with group 1 (negative controls). It also revealed a positive correlation with the incorporation studies. Significant increase in the CCPR and radiolabeled precursor incorporation (p <0.001) was observed in MNU-induced+TI fed group of animals (group 4) in all the three segments when compared with control diet fed normal (group 1) as well as MNU-induced (group 3) animals. This study therefore indicates a co-stimulatory effect of TI on MNU-induced colonic cell kinetics.

Ca2+ wave activity was monitored in the longitudinal (LM) layer of isolated murine caecum and proximal colon at 35 degrees C with fluo-4 AM and an iCCD camera. Both intracellular (within LM cells) and intercellular (also spreading from cell to cell) Ca2+ waves were observed. Intracellular Ca2+ waves were associated with a lack of muscle movement whereas intercellular Ca2+ waves, which were five times more intense than intracellular waves, were often associated with localized contractions. Several intracellular Ca2+ waves were present at the same time in individual LM cells. Waves in adjacent LM cells were not coordinated and were unaffected by TTX (1 microM) but were blocked by IP3 receptor antagonists xestospongin-C (Xe-C; 2 microM) or 2-aminoethyl diphenylborate (2-APB; 25 microM), and by ryanodine (10 microM). Caffeine (5 mM) restored wave activity following blockade with Xe-C. NiCl2 (1 mM) blocked intracellular Ca2+ waves, and nicardipine (2 microM) reduced their frequency and intensity, but did not affect their velocity, suggesting the sarcoplasmic reticulum may be fuelled by extracellular Ca2+ entry. Intercellular Ca2+ waves often occurred in bursts and propagated rapidly across sizeable regions of the LM layer and were blocked by heptanol (0.5 mM). Intercellular Ca2+ waves were dependent upon neural activity, external Ca2+ entry through L-type Ca2+ channels, and amplification via calcium-induced calcium release (CICR). In conclusion, intracellular Ca2+ waves, which may reduce muscle excitability, are confined to individual LM cells. They depend upon Ca2+ release from internal Ca2+ stores and are likely to be fuelled by extracellular Ca2+ entry. Intercellular Ca2+ waves, which are likely to underlie smooth muscle tone, mixing and propulsion, depend upon neural activity, muscle action potential propagation and amplification by CICR.

The large intestine of mammals has long been viewed as an osmoregulatory organ, and evidence suggests that fluid and solute transport mechanisms within the intestine are heterogeneous, varying depending on the particular segment involved. Variations in function are often matched by morphological correlates, but despite the widespread use of rabbit large intestine as an experimental model, there is a lack of knowledge about the cellular makeup and dynamics in the colonic mucosal epithelium. The presence of mitotic figures and immunohistochemical localization of proliferating cell nuclear antigen (PCNA) were used to identify the proliferative zone(s). Cellular migration patterns were determined through the use of the thymidine analog 5-bromo-2-deoxyuridine (BrdU) over a 24-, 48-, and 72-hr period. Apoptotic nuclei were identified utilizing terminal deoxynucleotidyl transferase d-UTP nick-end labeling (TUNEL). Both cecum and the initial portion of the proximal colon (P1) exhibited a proliferative zone at or near the crypt base, and migration proceeded upwards toward the surface epithelium lining the intestinal lumen, where apoptosis occurred Turnover time of crypt columnar cells was determined to be about 3 days; that of mucous cells was estimated to be about 5 weeks. Rabbit cecum and proximal colon P1 are similar in their cellular morphology and epithelial cell kinetics. In both, the major proliferative zone is located at or near the crypt base, from which crypt columnar cells migrate toward the lumenal surface epithelium over a period of 3 days. Goblet cell turnover rate is much slower than that of columnar cells., (Copyright 2001 Wiley-Liss, Inc.)

Changes in the frequency of endocrine cells are evidence of intestinal adaptation to germ-free (GF) status. Not only the distribution of these cells along the intestine, but also the differences in intracellular content of these regulatory peptides may be explored to explain functional and structural aspects of GF intestinal adaptation. Focusing on the endocrine L-cells, we analyzed the intracellular content of enteroglucagon (EG) and peptide YY (PYY) throughout the intestine of the 14 GF and 14 conventional (CV) mice by using immunohistochemistry and the supra-optimal dilution technique. The percentage of EG-immunoreactive cells, but not of PYY-immunoreactive cells stained at supra-optimal dilution was significantly higher in the proximal colon of GF mice than in the CV counterparts (P < 0.05). Since the content of co-stored PYY did not differ between GF and CV mice, the higher content of EG was compatible with a selective cellular response. Moreover, in the cecum of GF mice, the density of EG-immunoreactive cells was significantly higher than that of PYY-immunoreactive cells (P < 0.05). These results are consistent with preferential production of EG by L-cells at the expense of PYY in the proximal colon and in the enlarged cecum of GF mice. In addition, they may reflect the dynamics of the GF intestinal epithelium and/or be correlated with the higher serum levels of these peptides. The role of endocrine cells needs to be better studied in human and other experimental adaptative conditions in order to elucidate the regulatory mechanisms of intestinal functions.

The arrangement of the bundles of muscular fibers in the transition between the small and the large intestines was studied in 12 male adult crossbred swine by dissection, after immersion in 50% nitric acid solution. The connection between the small and the large intestines was formed by the association of the muscular tissue, the connective tissue and the fat tissue. The tunica muscularis of these organs was organized in thin thread-like bundles and wide ribbon-like bundles of variable width and thickness. The superficial bundles of the longitudinal stratum of the tunica muscularis of the ileum established the continuity with the large intestine; the deep bundles penetrated into the ileal papilla. The limit between the cecum and the ascending colon was externally marked by the sulcus cecocolicus dorsalis and ventralis. The connection between the cecum and the ascending colon was formed by bundles of muscular fibers coming from the ileum, and the taeniae ventralis, lateralis and medialis of the cecum. Some bundles of muscular fibers from the ascending colon and the cecum headed toward the sulcus cecocolicus. The median bundles of muscular fibers of the taenia ventralis of the cecum, near the termination of the ileum, were arranged to form a loop around the termination of the ileum, mixing with the musculature of the ascending colon at the level of this junction.

Morphological characteristics and cellular proliferation were investigated in the hindgut tissue of 25 pigs ranging from 5 to 261 days of age; the three youngest pigs were unweaned. Tissue samples were taken from the cecum and from the proximal, middle, and distal part of the colon. In the young pigs a high incidence of branched crypts was observed. During the first three to four months there was an increase in crypt height and proliferative activity, as determined by the mitotic count, as well as an increase in the mucin secretion, especially the sulfomucins. Distinct regional differences were observed between the four intestinal sites. In general, the crypts were deeper and more closely spaced and the turnover time was higher in the distal part of the colon as compared to the cecum and the proximal colon. Furthermore, a greater proportion of the mucins in the middle and distal part of the colon are acidic or sulfated as compared to the cecum, where the mucins are more of the neutral type. These regional and age-related differences in morphological characteristics of the hindgut in pigs may have significance for the etiology of intestinal infections.

The response of the intracellular pH (pHi, measured with BCECF) of the caecal and distal colonic epithelium of guinea pig and of monolayers of HT29 clone 19a cells on the addition of short-chain fatty acids (SCFA) was assessed. Addition of SCFA to the luminal side of these cells had no major effect on pHi, independent of whether the apical Na+/H+ exchange or the apical K+/H+ ATPase was inhibited or not. Addition of SCFA to the serosal side, on the other hand, caused a marked decrease of pHi, followed by an effective regulation back to basal values, and after removal of the acid, the cells became alkalinized. Intracellular pH is mainly regulated by mechanisms in the basolateral membrane. The basolateral Na+/H+ exchanger and the Cl-/HCO3- exchanger were mainly responsible for pHi regulation. Inhibition studies are consistent with a NHE-1 type Na+/H+ exchanger in the basolateral membranes. The apical Na+/H+ exchanger of caecal enterocytes and in HT29 cells, and the apical K+/H+ ATPase in the apical membrane of the distal colon have no or little influence on pHi regulation. The comparison shows that the HT29-19a cell line is an adequate model for studying pHi phenomena of hind gut epithelial cells.

Two experiments were conducted with neonatal pigs to determine the effects of feeding fructooligosaccharides on cecal and colonic microbiota, proliferation of cecal and colonic epithelial mucosa, and short-chain fatty acid concentrations in the cecum. Experiment 1 consisted of feeding neonatal pigs diets containing either 0 or 3 g fructooligosaccharies/L of formula for 15 days and then examining the large intestine for changes in cecal and proximal colonic microbiota; cecal pH; short-chain fatty acid concentrations; morphology of cecal, proximal, and distal colonic epithelial mucosa; gross necropsy; and histopathology. Supplementation with fructooligosacchariudes (FOS) did not alter cell counts of viable bifidobacterial organisms or total anaerobic microbiota, cecal pH, or concentrations of short-chain fatty acids. Cecal mucosal cell density and labeled cells increased with FOS consumption. Proximal colonic mucosal crypt height, leading edge, labeled cells, proliferation zone, and labeling index increased with FOS consumption. Distal colonic mucosal crypt height, leading edge, cell density, labeling index, and labeled cells increased with FOS consumption. Gross necropsy and histopathology found no significan lesions. In Experiment 2, neonatal pigs were fed diets containing either 0 or 3 g fructooligosaccharides/L of formula for 6 days. Fecal samples were collected on the first full day of feeding and on days 3 and 6 after initiation of feeding. On days 1 and 3, concentrations of bifidobacteria were similar between diets; however, on day 6, pigs consuming FOS tended to have greater numbers of bifidobacteria (p = 0.08). These data suggest dietary consumption of FOS will enhance bifidobacteria populations and prevent colonic epithelial mucosa atrophy in neonates fed an elemental diet.

Two experiments were conducted to determine if supplementing soluble fiber (fructooligosaccharide, xylooligosaccharide or gum arabic) to a semi-elemental diet would beneficially change cecal and colonic microbiota populations and enhance epithelial cell proliferation. Experiments 1 and 2 used identical dietary regimens; mice and rats were given free access to a powdered semi-elemental diet. Animals were assigned to one of the four following treatment groups: control, no supplemental dietary fiber, fructooligosaccharide, xylooligosaccharide and gum arabic. Dietary fiber was supplied via drinking water at 30 g/L. In Experiment 1 populations of Bifidobacteria and total anaerobic flora were enumerated from the contents of the cecum and colon of weanling mice. Consumption of fructooligosaccharide increased (P < 0.05) the concentrations of Bifidobacteria and the ratio of Bifidobacteria to total anaerobic flora. In Experiment 2 tissue from the cecum and distal colon of weanling rats was examined for morphological changes of the mucosa. Consumption of xylooligosaccharide increased (P < 0.05) cecal crypt depth and labeling index relative to the other three treatments. Consumption of gum arabic and the control diet increased (P < 0.01) cecal proliferation zone. Consumption of xylooligosaccharide and the control diet increased (P < 0.01) cecal cell density (number of cells in a vertical-half of the crypt). Distal colonic crypt depth was greatest (P < 0.05) in controls and rats fed fructooligosaccharide, intermediate in those fed gum arabic, and smallest in those fed xylooligosaccharide. These results suggest that fructooligosaccharide effectively stimulates growth of Bifidobacteria and xylooligosaccharide supports a modest enhancement of cecal epithelial cell proliferation.

In this work we have demonstrated the influence of a short chain fatty acid (acetate) on the number of enterochromaffin (EC) cells containing serotonin (5HT), at two different pH (pH 6.9 absorptive stimuli, and pH 2.9 secretory stimuli), infused into the colon during one hour. The number of EC cells decrease significatively, specially in the cecum with a solution of low pH (2.9). The action of piprenzepine in preventing this reduction demonstrated that was partly mediated by a cholinergic receptor mechanism. On the other hand, a decrease on the release of 5HT to the lumen was a observed under the influence of pirenzepine. We conclude that the short chain fatty acid acetate, at a low pH induces the release of serotonin through a cholinergic mechanisms indicated by the inhibition observed with antimuscarinic drug.

The in vitro metaphase arrest technique (crypt cell production rate-CPPR) has been used to measure human rectal mucosal proliferation. Study of preincubation times, dose response curves and lag phases suggest that a concentration of vincristine of 5 micrograms/ml and 16 hour preincubation with time point increments between 25 and 125 minutes give optimal conditions for measuring rectal mucosal proliferation. Twenty individuals had rectal CCPR repeated without intervention of any kind. Close correlation was found between the two values (r = 0.89 and P = 0.0001). The effect of polyethylene glycol bowel preparation was also studied in 35 subjects. There was good correlation (r = 0.66, P = 0.007). There was close correlation between rectal and caecal CCPR as measured in 20 patients who had colonoscopy (r = 0.72, P = 0.0003). The in vitro metaphase arrest technique is a useful parameter of rectal mucosal proliferation and may be used with confidence in a number of different clinical situations.

The pelvic flexure portion of the equine large colon is the proposed location of a pacemaker mechanism. This study was conducted to ascertain whether the distribution of certain putative neurotransmitters differs at the pelvic flexure compared to other sampling sites. Tissue samples were collected from the intestinal tracts of six horses. Serial sections from these samples were reacted with primary antisera specific for substance P, vasoactive intestinal polypeptide (VIP), methionine-Enkephalin, and calcitonin gene-related peptide (CGRP). The regional distribution of immunoreactive neuronal elements was uniform for each of the neuropeptides except VIP. Although neurons exhibiting VIP-like immunoreactivity were abundant throughout the colon, they were somewhat more plentiful near the apex of the pelvic flexure and the left dorsal colon. These neurons may participate in the initiation and propagation of the propulsive/retropulsive contraction waves, which emanate from this location and are believed to lend a sphincter-like capacity to the pelvic flexure. The submucosal plexus was replete with neurons with intense substance P and VIP-like reactivity. Reactive fibers left submucosal ganglia to project to the intestinal mucosa, reflecting a possible secretogogic role for these neurons. This role may be especially important for the horse as a hindgut fermenter. There were abundant methionine-Enkephalin and substance P-like reactive varicosities throughout the myenteric plexus, many of which established a pericellular plexus of varicose fibers. The abundance of these varicosities, which may correlate with a high degree of neuronal integration, did not vary regionally. These data may enhance our understanding of both normal colonic peristalsis and motility disorders caused by a depletion of these neuropeptides.

The crypt cell production rate (CCPR) is considered the most robust estimate of cell turnover. The CCPR was determined at various sites around the colon in the Wistar rat (using an in vivo technique) and in six healthy humans (using in vitro organ culture). In both the rat and human colon, the CCPR increased proximally from the rectum to the caecum. The caecum had a significantly higher cell turnover than any other site in the colon (p < 0.05 in the rat, p < 0.01 in humans, Student's t test). These findings are of interest when considering cellular proliferation studies in both inflammatory bowel disease and colorectal cancer, and draw attention to the importance of choosing a constant reference site for comparative studies. This is the first time the CCPR had been measured along the length of the human colon.

The possibility that VIP (Vasoactive intestinal peptide) could influence the enterochromaffin (EC) cell secretion of serotonin (5HT) and the action of VIP on the mast cell population of lamina propria were investigated in Wistar rat colon infused with a short chain fatty acid solution (sodium acetate), during a 1 h period. Under the action of an intravenous injection of synthetic porcine VIP, 14 micrograms/kg/h), the number of EC cells diminished significantly in the cecum and left colon, when compared to non-injected animals, both infused with a sodium acetate solution. At the same time, the number of mucosal mast cells in the crypts and lamina propria decreased significantly in the cecum. The postulate we put forward is that these VIP-induced changes are exerted through the stimulation of 5HT released from EC cells not only under normal physiological conditions but probably also under pathological conditions.

Cells isolated from the jejunum, cecum and colon of rats were used to study the oxidation of nutrients by quantifying the production of 14CO2 from 5 mmols/L 14C-labeled exogenous substrate. In colonic cells, the decreasing order of oxidation was as follows: butyrate greater than acetate greater than propionate, glucose and glutamine. Acetate and butyrate significantly suppressed oxidation of both glucose and glutamine. In cells taken from the cecum, butyrate was oxidized at a greater rate than all other substrates. Cells taken from the jejunum produced CO2 from exogenous substrates in decreasing order as follows: glutamine greater than glucose much greater than acetate, propionate and butyrate. Butyrate oxidation was significantly reduced in colonic cells by 3-hydroxybutyrate, and it was reduced in cecal cells by glucose. Comparisons among the three gut segments showed no differences in glutamine oxidation. Glucose oxidation was greater in cells taken from the colon than from the cecum or jejunum, which were similar. Butyrate and acetate were oxidized at higher rates in cells taken from the cecum and colon than in cells taken from the jejunum, and propionate was oxidized at a greater rate in cells taken from the colon than from the jejunum. These studies demonstrate that relative rates of substrate oxidation differ along the intestinal tract of rats.

In the rat, topographic and X-ray studies of the caecum and ascending colon, together with microscopic anatomic observations of the ileal papilla and caecocolonic valve, showed that junction structures contribute to prevent the backflow of the caecal content into the ileum and to direct the ileal content within the caecum whatever the various positions of the caecum in the abdominal cavity.

Variations in percentage labelling (LI) and mitotic activity (mitoses per crypt) have been studied over a 24-hr period in the epithelial cells and pericryptal fibroblast sheath (PCFS) of the small intestine, caecum, and colon of the mouse. All three tissues displayed clear, synchronized circadian rhythms in DNA-synthetic activity in both the epithelial cells and PCFS. Peak values were coincident within a tissue, but staggered between tissues. In the epithelial cells, peak mitotic values were found between 3 and 6 hr after the peak LI values. The low level of mitotic activity in the PCFS appeared to be synchronized with the rhythms in the adjacent epithelia. The epithelial cells of the lowest crypt third displayed the clearest circadian rhythms. However, the PCFS cells at all levels produced similar curves. A craniocaudal wave of proliferative activity is proposed.

The influence of luminal pH on the colonic epithelial cell cycle was studied by means of dietary modification to produce acidification of colonic contents. Forty male Sprague-Dawley rats were fed one of four diets: defined fiber free, fiber free diluted by either 100 g/kg lactulose or sorbitol, or 25 g/kg of MgSO4. After 2 wk, in vivo pH measurements were recorded at laparotomy under ether anesthesia in the cecum and proximal, mid-, and distal colon. Epithelial cells, isolated from these same regions, were measured for DNA content with flow cytometry in order to determine cell cycle stage. The pH values in all intestinal regions were highest with MgSO4 and then fiber free, sorbitol, and lactulose, the most acidic. In contrast, the percentage of cells in S phase (actively synthesizing DNA) was highest with lactulose and least with MgSO4. There was a significant inverse correlation between luminal pH and percent cells in S phase in the cecum (P less than 0.01), proximal colon (P less than 0.05), and distal colon (P less than 0.01). These results show that acidification of colonic contents by diet modification leads to increased epithelial cell proliferation.

The authors report the time of appearance, morphology and topographic distribution of gastrin/cholecystochinin- (G/CCK-), somatostatin- (SRIF-), neurotensin- (NT-), motilin- (MO-) and substance P-like immunoreactive (SP-LI) elements during embryonic and postnatal development, in ileum, caeca and colon of chick embryos (from 8 days of incubation to hatching), newborn chicks (up to 15-days old) and adult chickens. In the ileum, G/CCK-LI and SP-LI cells appeared on day 11, the others on about day 13. In the caeca the first cells of all types were seen from about day 17. In the colon, NT-LI cells appeared early, on day 9, SP-LI and occasional SRIF-LI cells from day 13 on and MO-LI and G/CCK-LI only from day 17. In the ileum all the cells studied were present, in the caeca and colon they were extremely scarce, apart from NT-LI cells which were more numerous. In the prenatal stages, SP-LI was found only in epithelial cells; after hatching, it was also present in metasympathetic nerve elements.

Influence of hindgut bypass on the epithelial cell production rates of the small and large intestine was tested in rats. Hindgut bypass decreased cryptal cell production rates of the jejunum, ileum, cecum, and distal colon by 40-65%. These results suggest that the entrance of ileal effluent into the hindgut and, presumably, the resultant microbial production of short-chain fatty acids are required to maintain the normal epithelial cell proliferation not only in the hindgut but also in the small intestine.

Light microscopy, morphometry and scanning electron microscopy (SEM) were used to examine the mucosal morphology of seven intestinal specimens (three from the small intestine and four from the large intestine) from two horses not subjected to surgery and three horses one year after sham-operation for colon resection. Qualitative and quantitative evaluation revealed similar morphology for all horses except that the unoperated horses had significantly (P < 0.05) fewer goblet cells in the crypts of the large intestine. In the small intestine, SEM demonstrated that villus shape varied and ranged from coned to broad and flat. In the large intestine, SEM of the right ventral and dorsal colon revealed more prominent crypt openings than in the caecum. The small colon contained the most prominent crypt openings with the least variation among horses. In the small intestine, light microscopy and morphometry revealed a greater number of goblet cells and a decreased width to the enterocytes in an aboral direction from the cranial jejunum to the ileum. In the large intestine, the crypt area and crypt depth increased significantly and the intercrypt area decreased significantly in an aboral direction from the caecum to the small colon (P < 0.05). In comparison to the normal morphology, horses one year after extensive large colon resection had significantly greater intercrypt area in the caecum and remaining large colon.

A microdissection technique used previously in the small intestine has been adapted for the rat colon in order to determine morphometric data and to calculate surface parameters in various proximal and distal segments (caecum, ascending and descending colon, rectum) of normal colonic mucosa. From the caecum to the rectum there was a gradual significant decrease in the diameter of the middle portions of the crypts, in the number of crypts per unit area and the inner intestinal circumference, as well as in the total mucosal surface per mm intestinal length and the surface of all crypts per unit area. However, there was an increase in the surface of the lining epithelium between the crypts from the caecum to the rectum. These findings are of importance when interpreting alterations in colonic function in terms of segmental changes in the mucosal architecture.

The peculiarities of enterocyte proliferation in the duodenum, jejunum, ileum, caecum, ascending, transverse and descending colon in the rat were studied by different methods of analysis of cell population kinetics (percentage-labelled mitosis curves, cumulative labelling curves, distribution of labelling index curves, etc.). The majority of proliferating cells in the small intestine are homogenous, as far as mitotic cycle mean duration (11-12 hrs) is concerned. All proliferating cells in all the zones of the colonic crypts and the bottom of the small intestine crypts are divided into subpopulations, having different mean durations of the mitotic cycle. It is suggested that, in the crypt bottom in all intestines as well as the crypt's maximum proliferation zones in most of the colonic segments, a considerable fraction of cells has a very long mitotic cycle or enters resting phase R1. The average value of the mean durations of the mitotic cycle of all colonic enterocyte subpopulations is 18-22 hours. On the basis of the authors' findings and literature data, a model for the enterocyte life cycle is proposed, according to which the cell flux is branched during the mitotic cycle and the crypt develops from a stem enterocyte population located at its bottom.

The long-term effects of fermentable fibers on colonic luminal pH and the epithelial cell cycle were compared in 50 male Sprague-Dawley rats fed either a defined basal fiber-free diet or the basal diet supplemented with 10% pectin, cellulose or guar or with 20% oat bran. After 8 mo, in vivo pH measurements revealed that acidification of luminal contents occurred in the cecum and in mid and distal colons of rats fed fiber-supplemented diets when compared with the fiber-free controls (P less than 0.05). Pectin and guar produced the greatest acidification of luminal contents, the largest increase in cecal surface area and the highest percentage of colonic cells in S-phase, as measured by flow cytometry. In the proximal colon of the pectin group 9.2 +/- 0.5% of cells were in S-phase (6.3 +/- 0.8% with the fiber-free group) (P less than 0.05) and in the distal colon of the guar group 10.9 +/- 1.4% were in S-phase (7.1 +/- 0.5% with the fiber-free group) (P less than 0.05). Even though the most fermentable fibers produced the greatest mitogenic response, there was no site-specific correlation between pH and mucosal cell growth except in the cecum. This suggests that fibers may act as colon cell growth factors by some mechanism other than extracellular pH changes.