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Absorption of short-chain fatty acids (SCFA) and ammonia implies considerable fluxes of protons across the epithelium of the large intestine. Efficient regulation of intracellular pH (pH(i)) is therefore essential in these cells. The aim of the present study was to examine the effects of SCFA and of ammonia on pH(i), on pH(i) regulation and to characterize the mechanisms involved in pH(i) regulation in surface enterocytes of the guinea-pig caecal and colonic mucosa. Intact epithelia from the caecum and the distal colon were mounted in a microperfusion chamber. pH(i) was measured by fluorescence microscopy using 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF). Addition of SCFA or ammonia to the serosal side changed the enterocyte pH(i) markedly, whereby ammonia caused larger changes in pH(i) than SCFA. In contrast, addition of SCFA to the mucosal solution had no effect on pH(i) and ammonia increased pH(i) only slightly. Basolaterally located pH(i) regulation mechanisms, Na(+)-H(+) exchange and Cl(-)-HCO(3)(-) exchange, are involved mainly in returning pH(i) to normal values. It is concluded that, due to apparently lower permeability of the apical membranes, the caecal and colonic epithelium is protected against pH(i) disturbances caused by the naturally high luminal SCFA and NH(3) concentrations. The major regulation mechanisms of pH(i) are located in the basolateral membrane of the enterocytes.

Short-chain fatty acids (SCFA) are produced by microbial fermentation in the hindgut in considerable amounts. Most of the anions in hindgut contents are SCFA, mainly acetate, propionate and butyrate. SCFA are rapidly absorbed. Mechanisms involved in the transepithelial transport are discussed. Besides the contribution to the overall energy metabolism of animals or men, SCFA have a number of further important effects on the colonic mucosa. Factors affecting the pH of compartments in the mucosa, cell swelling, stimulation of mucin release and of mucosal blood flow are mentioned. Controversial reports are known on the role of SCFA in the metabolism of colonocytes. In spite of the conflicting opinions on the interaction between SCFA metabolism and the development of colitis ulcerosa, diverticulosis and colorectal cancer seems to exist. The obscure differences between the effects of SCFA on cell proliferation, differentiation and apoptosis of colonocytes in vivo and in vitro indicate that besides direct effects of SCFA systemic effects such as neural and humoral factors are of crucial importance. The opposing effects of SCFA on proliferation and apoptosis in normal colonocytes and in colonic cancer cells may open possibilities for prevention and/or therapy of patients with colonic diseases.

P‐glycoprotein (MDR1), that confers multidrug resistance in cancer, and the cystic‐fibrosis transmembrane‐conductance regulator (CFTR), that is causative defective in cystic fibrosis, belong to the family of ATP‐binding transport proteins. The expression of MDR1 and CFTR in human epithelial tissues and the cell lines T84 and HT29 was estimated by primer‐directed reverse transcription (RT) and subsequent monitoring of the kinetics of cDNA product formation during the polymerase chain reaction (PCR), MDR1 mRNA was found in high levels, 15–50 amol mRNA/μg RNA, in the intestine, kidney, liver and placenta, and in low levels, 0.2 amol/μg RNA, in respiratory epithelium. Large amounts of CFTR mRNA were measured in the gastrointestinal tract, whereas the kidney, as the phenotypically normal organ, and the lung, as the most severely affected organ in cystic fibrosis, both contained low amounts, 3 amol CFTR/μg RNA. CFTR transcript levels of 1–5 amol/μg RNA were determined in lymphocytes and lymphoblast cell lines, suggesting that lymphoblasts are an accessible source for the study of the molecular pathogenesis of cystic fibrosis. When transcripts were scanned by overlapping RT/PCR analyses, only transcript of expected size was detected for MDR1 mRNA, whereas variable in‐frame deletions of either exon 4, 9 or 12 were observed in CFTR mRNA. The complete loss of single exons was seen at proportions of 1–40% in all investigated tissues and cell lines with large donor‐to‐donor variation. Exons 9 and 12 of the CFTR gene encode parts of the evolutionarily well‐conserved first nucleotide‐binding fold including the two Walker motifs. Alternative splicing may give rise to various CFTR forms of different function and localization.