SUMMARY1. Sugar-containing diets chosen not to affect intestinal structure or enterocyte turnover have been fed to mice previously maintained on a low carbohydrate diet in order to determine their ability to induce disaccharidase enzymes in the small intestine.2. Glucose-, fructose-and 3-0-methyl-glucose-containing diets increased sucrase and maltase but not lactase activities in mouse jejunal homogenates. These effects were either absent or negligible in more distal regions of the small intestine.3. Placing mice on glucose-, fructose-or 3-0-methyl-glucose-containing diets was further shown, by quantitative cytochemistry, to cause a 1-6-, 2-6-and 3*2-fold increase in the initial rate at which a-glucosidase activity (sucrase + maltase) appeared in the brush-border membrane of developing enterocytes.4. The time during which a-glucosidase activity increased in enterocyte brushborder membranes fell from 30 h for low carbohydrate fed mice to 21, 19 and 17 h in mice fed glucose, fructose and 3-0-methyl-glucose respectively. Change of diet had no effect on the kinetics of lactase expression by developing enterocytes.5. Maximal a-glucosidase activity detected in enterocyte brush-border membranes is equal to RT, where R is the initial rate of enzyme appearance and T is the time during which this rate operates. The ability of sugars to increase R selectively, but only at the expense of T, defines unexpected limits to the capacity of enterocytes to adapt to changes in luminal nutrition.6. The above results are discussed in relation to other aspects of enterocyte differentiation recently subjected to quantitative analysis. The need to standardize other aspects of intestinal physiology and redefine the energy content of diets containing non-metabolizable substrates in this type of work is also emphasized.

The enterocyte undergoes sequential changes in its structure and function as it migrates rapidly from the small intestinal crypts to the villus tip. The mechanisms by which these changes are regulated "in tune" with ontogenic and dietary changes in the luminal environment are currently under investigation. This study has employed oligonucleotide probes to follow the expression of the lactase-phlorizin hydrolase (LPH) and Na(+)-glucose cotransporter (SGLT1) genes in rabbit small intestine using quantitative in situ hybridisation histochemistry. The profiles of LPH mRNA and SGLT1 mRNA accumulation along the crypt-villus axis were found to be very similar. Although mRNA was undetectable in the crypt. LPH and SGLT1 mRNA levels rose rapidly at the crypt-villus junction, reaching a maximum between 210 microns and 330 microns above this point. Further up the villus the level of mRNAs declined. SGLT1 mRNA was present in all small intestinal segments (duodenum, jejunum and ileum), whereas LPH mRNA was absent from the ileum. LPH activity rose and fell in conjunction with mRNA, but SGLT1 activity was greatest at the villus tip where mRNA levels were considerably reduced. These data have been used to discuss the genetic regulation of enterocyte differentiation and function.

The physiological mechanisms that regulate epithelial gene expression during enterocyte migration and differentiation are still poorly understood. The present study has used a combination of quantitative in situ hybridisation, immunohistochemistry and enzyme cytochemistry to examine epithelial cell differentiation in rabbit small intestine. We have measured and compared the levels of mRNA and enzyme activity of the enterocyte brush border markers alkaline phosphatase, amino-peptidase N and lactase in normal villus epithelia and in epithelial cells exposed directly to the Peyer's patch immune environment. All three genes appeared to be expressed in parallel, but in each epithelial population examined, the pattern of gene expression was different. The level of these mRNAs was markedly reduced in Peyer's patch-associated epithelia, this being most pronounced in the follicle-associated epithelium, compared with normal villi. The activities of alkaline phosphatase and aminopeptidase N approximated the expression of their genes, whereas additional post-transcriptional events were shown to clearly contribute to the level of lactase activity in these tissues. These findings demonstrate that the reduced brush border hydrolase activity in Peyer's patch tissue that has been observed previously, is due to a down-regulation of epithelial gene expression in this location. These observations have been used to discuss epithelial differentiation in Peyer's patch tissue and the possible role of local immune factors in regulating such events.

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