Biomedical subjects

D J Winton

Publications and source records attributed to D J Winton.

23 records · Page 2Linked to original sources

PubMed1988

A clonal marker induced by mutation in mouse intestinal epithelium.

A cellular marker for individual somatic cells and their clonal descendents would be a valuable tool for the investigation of cell lineages and clonal organization in developing and in renewing tissues. Such markers have been developed in Drosophila, but (apart from mutant melanocytes in retinal pigmented epithelium) not so far in mammalian tissues. We report here the development of a mutation-induced marker in mice heterozygous at the Dlb-1 locus which determines the expression of binding sites for the lectin Dolichos biflorus agglutinin (DBA) in intestinal epithelium. We show that this marker can be used to study the clonal organization of adult intestinal epithelium, and to mark descendent clones arising during development. The method can in principle be extended to any other suitable markers which can be obtained in a heterozygous state, including markers generated in transgenic animals.

Animals↗

PubMed1988

Quantitative electron microscopy of carcinogen-induced alterations in hepatocyte rough endoplasmic reticulum. I. Chronic effect of 3'MeDAB and short-term effects of azo dyes of different carcinogenic potentials.

A new method has been developed for the quantitative analysis of an ultrastructural change in hepatocyte rough endoplasmic reticulum (ER) that is induced by liver carcinogens. Hitherto only subjective observations of this alteration had been made. Male inbred Leeds rats were fed a diet containing 0.06% of the carcinogenic azo dye 3'-methyl-4-dimethyl-aminoazobenzene (3'MeDAB). Groups of treated rats, together with untreated controls, were sacrificed after 10 days, 4 weeks, 10 weeks and 17 weeks. Samples of liver tissue from each animal and from 10 3'MeDAB-induced hepatocellular carcinomas (HCC), were examined by electron microscopy. A quantitative study was carried out to investigate the effect of chronic exposure to 3'MeDAB upon the state of aggregation of the hepatocyte rough ER into parallel arrays. As early as 10 days after the start of treatment, the rough ER showed a highly significant degree of disaggregation: the mean number of ER cisternae per array fell from the control value of 6.20 to 3.73. This change was sustained throughout the experiment. At 17 weeks, comparison of the mean array size in the HCC cells with that in the surrounding hepatocytes revealed a further significant decline, from 3.46 to 2.12. Further groups of rats were fed other azo dyes for 4 weeks and subjected to the same assay. The carcinogens 4'-methyl-4-dimethylaminoazobenzene (4'MeDAB) and N,N-dimethyl-4-amino-N-acetyl-N-monomethyl-4-aminoazobenzene (DAAMAB) resembled 3'MeDAB with respect to the degree of rough ER disaggregation they induced. In contrast, the non-carcinogen 3'-trifluoromethyl-4-dimethylaminoazobenzene (3'TFMeDAB) had no such effect, while the weak initiator 2-methyl-4-dimethylaminoazobenzene (2MeDAB) induced disaggregation to a lesser degree than the strong carcinogens. At least with the azo dyes used in this study, the extent of rough ER disaggregation appears to be related to hepatocarcinogenesis.

Animals↗

PubMed1988

Development of the pattern of cell renewal in the crypt-villus unit of chimaeric mouse small intestine.

We have previously shown that the epithelium of each adult intestinal crypt in chimaeric mice is derived from a single progenitor cell. Whether the crypts are monoclonal from the outset-that is, are formed by the proliferation of a single cell-or whether their formation is initiated by several cells was not known. Here we report that many crypts contain cells of both chimaeric genotypes in the neonatal period indicating a polyclonal origin at this stage of morphogenesis. The cellular organization of the early neonatal crypt is therefore different from that of the adult crypt, which includes a zone of 'anchored' stem cells above the crypt base. Within 2 weeks, however, the crypt progenitor cell and its descendants displace all other cells from the crypt and the crypt attains monoclonality. The distribution of enterocytes on chimaeric villi in the neonate shows a mottled pattern of mosaicism which is progressively replaced by coherent sheets of cells from the crypts, and within two weeks the orderly adult clonal pattern is established.

Animals↗

PubMed1988

Effect of fasting on aggregation of hepatocyte rough endoplasmic reticulum in adrenalectomized and 3'MeDAB-treated rats: quantitative electron microscope study.

A method for the quantitative analysis of the degree to which hepatocyte rough endoplasmic reticulum (ER) is arranged into parallel arrays was used to study the effect of fasting on rough ER aggregation in rat liver cells following either bilateral adrenalectomy or administration of the carcinogenic azo dye 3'-methyl-4-dimethylaminoazobenzene (3'MeDAB). One group of male inbred Leeds strain rats was subjected to bilateral adrenalectomy (ADX): after 1 week half of the animals were fasted for 24 h whereupon the whole group was killed. A second group of rats, fed for 4 weeks on a diet containing 0.06% of the carcinogenic azo dye 3'MeDAB, was similarly divided into two groups that were killed either with or without a prior 24-h fast. Untreated control groups of rats, both fasted and unfasted, were also killed. A quantitative electron microscope study was carried out to investigate the effect of each treatment on the degree to which the hepatocyte rough ER was aggregated into parallel arrays. ADX alone was without effect but caused a dramatic fall in rough ER aggregation when combined with fasting. At least as great an effect was induced by 3'MeDAB, with or without fasting, while fasting alone had a significant but much more modest effect than either ADX or the carcinogen. Thus, two disparate treatments induced morphologically identical responses in hepatocyte rough ER. The implications of this are discussed in terms of known interrelations between glucocorticoids and chemical carcinogenesis in the rat liver.

Adrenalectomy↗

PubMed1987

Fine structural alterations induced in the rat hepatocyte by adrenalectomy: influence of deoxycorticosterone acetate or fasting.

Electron microscopy was used to investigate the changes induced by bilateral adrenalectomy (ADX) in rat hepatocytes. Animals were killed at 1 week post-ADX, either with or without a prior 24 h fast. Further studies were carried out at 4 to 5 weeks after ADX to investigate the effect of administering deoxycorticosterone acetate (DOCA) for 4 weeks on the ADX-induced alterations. At 1 week, fasting had a profound effect on the response of hepatocytes to ADX in that either fasting or ADX alone caused proliferation of smooth endoplasmic reticulum (ER) and reduced glycogen but did not alter the rough ER, while ADX together with fasting resulted in the total depletion of glycogen, the virtual absence of smooth ER and the disaggregation of the rough ER into single cisternae. In contrast to the changes seen at 1 week after ADX in unfasted animals, at 4 weeks the hepatocytes contained increased glycogen in large, dense areas and had a scanty smooth ER, suggesting that some endogenous source of steroid hormones active in carbohydrate metabolism becomes available in male ADX rats by this time. This interpretation was consistent with the finding that DOCA-treatment of ADX rats prevented the glycogen accumulation seen at 4 weeks and also induced some smooth ER proliferation.

Adrenalectomy↗