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M Kedinger

Publications and source records attributed to M Kedinger.

At least 91 records · Page 5Linked to original sources

Developmental pattern of brush border enzymes in the human fetal colon. Correlation with some morphogenetic events.

The present study is concerned with a multilevel approach to human colon organogenesis, involving scanning and transmission electron microscopy together with brush border enzymology. The results emphasize the particular developmental pattern of sucrase activity which appears towards 11 weeks, increases at 14 weeks, begins to decrease around 28 weeks and disappears totally at term. In contrast, other enzymes like aminopeptidase and alkaline phosphatase persist in the adult colon. The correlation, in the fetal large bowel, of enzyme activities and villus structures similar to those found in the small intestine is discussed.

Alkaline Phosphatase↗

Effects of human fetal gastroenteric mesenchymal cells on some developmental aspects of animal gut endoderm.

Human intestinal and gastric mesenchymal cells were associated with chick and rat intestinal endoderm in order to test their species-specific capacity on epithelial differentiation. Primary cell cultures were established from human intestinal and gastric mesenchyme. Animal intestinal endoderms were associated with both cell types, grafted in ovo and allowed to develop for 12 days. The morphologic and enzymatic differentiation of the recombinants demonstrated two types of inductive properties exerted by human fetal intestinal and gastric mesenchymal cells, respectively. Firstly, human intestinal mesenchymal cells triggered intrinsic developmental capacities in chick and rat endoderm, i.e. enhanced structural brush-border maturation in both species and precocious sucrase induction in rat endoderm. Secondly, human gastric mesenchymal cells provoked the partial conversion of chick intestinal endoderm into gastric structures. Such properties were not found in homologous animal mesenchymes.

Alkaline Phosphatase↗

Early organogenesis of human small intestine: scanning electron microscopy and brush border enzymology.

Human small bowel early organogenesis was studied by scanning electron microscopy and found to be correlated to brush border enzymology. The appearance of the brush border enzymes sucrase, lactase, and aminopeptidase (measured in a purified apical membrane fraction) coincides with the first outgrowth of villi (eight weeks). Alkaline phosphatase was detected at seven weeks. The content of these enzymes furthermore increased up to the 14th week when both sucrase and aminopeptidase activities were comparable with adult values.

Alkaline Phosphatase↗

Sucrase-isomaltase: a marker of foetal and malignant epithelial cells of the human colon.

The presence of sucrase-isomaltase (SI), a glycoprotein hydrolase normally restricted to the brush border membrane of the enterocytes of the small intestine, was investigated in tumours which developed in nude mice inoculated with six human colon carcinoma cell lines (HT-29, Caco-2, HRT-18, HCT-8R, SW-480, and CO-115). Foetal and normal adult human small intestines and colons were used as controls. SI was studied by (1) immunofluorescence with rabbit antibodies raised against purified human small intestine SI; (2) polyacrylamide gel electrophoresis and immunoblotting; and (3) determination of the enzyme activity. SI was antigenically present, and enzymatically active, in all the tumours derived from Caco-2 and HT-29 cells. The presence of the enzyme was associated with that of typical brush borders at transmission electron microscopy examination. SI was absent from the tumours developed with the other four cell lines, as well as from the normal adult colon mucosa. SI was also present and active in the colons of mid-gestation foetuses, ranging in ages between 20 and 28 weeks; it was absent from the colons of late-gestation foetuses. The presence of SI in tumours derived from two cell lines suggests that this enzyme is a marker, so far unsuspected, of certain human colon cancers, and that the differentiation pattern of these particular cancers closely resembles that of the foetal colon.

Animals↗

Role of glucocorticoids on the maturation of brush border enzymes in fetal rat gut endoderm.

Heterospecific recombinants between fatal rat intestinal endoderm and chick mesenchyme, and also undissociated fetal rat intestine, were submitted to different hormonal environments. The present study shows that exogenously-supplied dexamethasone in organ culture, like endogenous hormones provided by the adult rat (grafting experiments) led to similar qualitative and quantitative results, i.e., a 9-fold stimulation of maltase and a precocious induction of sucrase activity in comparison with an hormonal conditions.

Animals↗

Inductive properties of fibroblastic cell cultures derived from rat intestinal mucosa on epithelial differentiation.

The present study represents a first attempt to elucidate the regulatory properties displayed by the non-epithelial portion of the intestinal mucosa, growing as fibroblasts in monolayer cultures. Thus, we compared the inductive action of 6-day suckling rat duodenal fibroblasts with that displayed by chick embryonic intestinal mesenchyme on the heterotypic cytodifferentiation of 5 1/2-day chick embryonic gizzard endoderm. The latter, isolated by 0.03% collagenase, was surrounded by intestinal intramucosal fibroblastic cell sheets. As control experiments, fibroblastic cells derived from the intestinal muscle or from 20-day fetal rat skin and lung were used. Every type of association was grafted into the coelomic cavity of 3-day chick embryos for 11 to 12 days, a system providing their vascularization and growth. The results clearly demonstrate that the mucosal fibroblastic cells of rat intestine were as potent as embryonic intestinal mesenchyme in inducing brush-border enzymes like sucrase and maltase, in conformity with an induced intestinal morphology. In contrast, the control fibroblastic cells were completely ineffective.

Alkaline Phosphatase↗

Organ culture of suckling rat intestine: comparative study of various hormones on brush border enzymes.

Jejunal mucosa of 6 d-old rats were cultured for 24 and 48 h in the presence of thyroxine, insulin, pentagastrin, glucagon, epidermal growth factor (EGF) or dibutyryl-A-3:5-MP cyclic with or without dexamethasone (DX). The enzymes were assayed on the purified brush borders. The various agents added alone to the basic culture medium had no effect with the exception of DX on the levels of enzyme activities. Dexamethasone alone induced sucrase, stimulated maltase, and protected other brush border enzyme activities (aminopeptidase, lactase, and alkaline phosphatase). When added to DX-supplemented medium, only the following factors modified the levels of enzymatic activities observed with DX alone. Insulin (10(-6) M) increased maltase, alkaline phosphatase, and lactase activity to a greater extent than DX at 24 h culture, the effect being maintained at 48 h on alkaline phosphatase only. At 48 h culture, both EGF (10(-8) M) and dbcAMP (10(-3) M) decreased DX-induced sucrase activity. The latter agent also depressed DX-stimulated aminopeptidase activity.

Alkaline Phosphatase↗

Control of brush border enzymes by dexamethasone in the fetal rat intestine cultured in vitro.

The effect of dexamethasone (DX) on the prenatal maturation of rat intestinal brush border enzymes was studied in organ culture. Jejunal segments were explanted daily from day 17 of gestation until birth, as well as at different postnatal stages until day 6; they were cultured for 48 h with or without DX (8 X 10(-8) M). Enzymatic activities were analyzed on brush border membranes purified from cultured intestines and were compared with values from uncultured specimens. The results showed that DX elicited (a) a precocious induction of sucrase activity in the jejunum explanted from 19 days of gestation onward, reaching a peak value when taken at birth; (b) a stimulation of maltase activity in the segments explanted as soon as day 18, leading to maximal values when taken at day 20, the stage at which the stimulated activity reached a 6.5-fold increase over the baseline activity; and (c) an increase of lactase activity comparable to that occurring in utero. As opposed to this, DX has no specific action on alkaline phosphatase and aminopeptidase activities. The present data indicate that glucocorticoids directly and specifically influence the prenatal maturation of some brush border enzymes in the mammalian gut.

Alkaline Phosphatase↗

[Permissive effects of human colonic cancer (HT-29 and Caco-2) on intestinal smooth muscle differentiation].

Human colonic carcinoma cells (lines HT-29 and Caco-2) were associated with 5 1/2 day-old chick embryonic intestinal mesenchyme and grafted for 11 days into 3 day-old Chick embryos. In these conditions, the cancer cells exert on the mesenchyme a supportative effect leading to its differentiation into muscular layers. The results provide additional evidence that HT-29 and Caco-2 cells exhibit common properties to fetal and normal intestinal cells.

Animals↗

Organogenetic potentialities of rat intestinal epithelioid cell culture.

The aim of the present study was to investigate the mesenchymal influence on cultured epithelioid cells originating from an already differentiated intestine. Epithelioid cell cultures of 6-day-old suckling rat intestine were established by sequential trypsinizations of the mucosa. Embryonic intestinal monolayers of quail cells (13 days) were used as control because of their natural cell marker. Six to thirty days after plating, both types of epithelioid cells were associated in heterospecific combination with 5 1/2-day-old chick embryonic small intestinal mesenchyme, after removal of the endoderm by collagenase treatment. In order to test the differentiation capabilities of the associations, they were grafted for 10-12 days into 3-day-old chick embryos. The results show that in such an in vivo culture system, the chimeric associations gave rise to well differentiated intestinal structures indicating that the epithelioid cell cultures derived from late embryonic or neonatal intestine will go through organotypic differentiation when recombined with an appropriate mesenchyme.

Animals↗

Comparative in vivo and in vitro effect of mono- and disaccharides on intestinal brush border enzyme activities in suckling rats.

Suckling rats were bottle fed during 48 h with a basic diet enriched with different mono- or disaccharides. In parallel, explants of intestinal mucosa were cultured in vitro for 48 h in the presence or in the absence of a synthetic glucocorticoid (dexamethasone) and with or without different carbohydrates. In both studies, enzyme activities were assayed on purified brush border membranes. From these combined in vivo and in vitro investigations it appeared that (1) there is a highly specific stimulation of mono- and disaccharides on the corresponding brush border disaccharidases: glucose, fructose, sucrose on sucrase and maltase activities, fructose being generally the most potent activator. Other brush border enzymes were not modified by the dietary carbohydrates. (2) These sugar-mediated effects were obtained only in the presence of glucocorticoids. This hormone alone induced the appearance of a slight sucrase activity and provoked a stimulation of maltase activity. The results show clearly that glucocorticoids are necessary to induce sucrase activity, but that the level of this activity is under the strict dependence of the dietary carbohydrates.

Alkaline Phosphatase↗

Effect of sucrose refeeding on disaccharidase and aminopeptidase activities of intestinal villus and crypt cells in adult rats. Evidence for a sucrose-dependent induction of sucrase in the crypt cells.

In order to gain more insight into the adaptative mechanism of intestinal enzymes to dietary factors in rats, modifications in the activities of disaccharidases and aminopeptidase were measured after refeeding of a 70% solution of sucrose for 15 h following a 2-day fast. Mature epithelial cells from the villus and immature cells from the crypt were isolated after sequential removal of the cells along the villus-crypt axis. Synthesis of brush border disaccharidases was determined by measuring [3H]valine incorporation into proteins. 1. In the whole mucosa, a highly significant increase in sucrase and maltase activities and a significant drop in aminopeptidase activity was observed in the brush border membranes after sucrose refeeding. 2. Stimulation of sucrase and maltase activities in sucrose refed rats was produced mainly in the immature cells of the crypt and lower villus compartment. 3. After separation of the brush border proteins by SDS gel electrophoresis from villus and crypt cells of sucrose refed rats, major incorporation of the radioactive precursor occured in the protein bands corresponding to sucrase and maltase activities of the lower villus and crypt cell brush borders. These findings demonstrate that sucrase stimulation by sucrose occurs mainly in the immature epithelial cells and that the substrate induces de novo synthesis of sucrase molecules.

Animals↗

[Islet cell transplantation and diabetes (author's transl)].

It is now apparent that transplantation of islet cells is biologically feasible in experimental animals utilizing adult neonatal or fetal islet cells and employing many different sites for implantation of the islet cells. It is reasonable to expect that with successful acquisition of new basic information in immunology, endocrinology, metabolism and developmental biology islet cell transplantation should be used as a therapeutic approach to human diabetes.

Animals↗