Rat Jejunal Epithelium Research Articles

Cholera toxin perturbs the paracellular barrier in the small intestinal epithelium of rats by affecting claudin-2 and tricellulin

Cholera toxin is commonly known to induce chloride secretion of the intestine. In recent years, effects on epithelial barrier function have been reported, indicating synergistic co-regulation of transporters and tight junction proteins. Our current study focused on the analysis of cholera toxin effects on transepithelial resistance and on tight junction proteins, the latter known as structural correlates of barrier function. Ligated segments of the rat jejunum were injected with buffered solution containing cholera toxin (1μg/ml) and incubated for 4h. Subsequently, selfsame tissue specimens were mounted in Ussing chambers, and cholera toxin (1μg/ml) was added on the apical side. Transepithelial resistance and permeability of sodium fluorescein (376Da) were analyzed. Subsequently, tissues were removed, expression and localization of claudins were analyzed, and morphological studies were performed employing transmission electron microscopy and confocal laser scanning microscopy. Cholera toxin induced a marked decrease in transepithelial resistance in the rat jejunal epithelium and an increase in paracellular permeability for sodium fluorescein. Immunoblotting of tight junction proteins revealed an increase in claudin-2 signals, which was verified by confocal laser scanning immunofluorescence microscopy, and a decrease in tricellulin, whereas other tight junction proteins remained unchanged. Transmission electron microscopy showed a reduction in the number of microvilli after incubation with cholera toxin. Moreover, cholera toxin led to a widening of the intercellular space between enterocytes. In accordance with the commonly known prosecretory effect of cholera toxin, our study revealed a complementary effect on small intestinal barrier function and integrity, which might constitute a pathomechanism with high relevance for prevention and therapeutic approaches.

Effects of Chronic 137Cs Ingestion on Barrier Properties of Jejunal Epithelium in Rats

Environmental contamination by 137Cs is of particular public health interest because of the various sources of fallout originating from nuclear weapons, radiological source disruptions, and the Chernobyl disaster. This dispersion may lead to a chronic ecosystem contamination and subsequent ingestion of contaminated foodstuffs. The aim of this study was to thus determine the impact of a chronic ingestion of low-dose 137Cs on small intestine functions in rats. The animals received 150 Bq per day in drinking water over 3 mo. At these environmental doses, 137Cs contamination did not modify the crypt and villus architecture. In addition, epithelial integrity was maintained following the chronic ingestion of 137Cs, as demonstrated by histological analyses (no breakdown of the surface mucosa) and electrical transepithelial parameters (no change in potential difference and tissue conductance). Furthermore, cesium contamination seemed to induce contradictory effects on the apoptosis pathway, with an increase in the gene expression of Fas/FasL and a decrease in the apoptotic cell number present in intestinal mucosa. No marked inflammation was observed following chronic ingestion of 137Cs, as indicated by neutrophil infiltration and gene expression of cytokines and chemokines. Results indicated no imbalance in the Th1/Th2 response induced by cesium at low doses. Finally, evaluation of the functionality of the jejunal epithelium in rats contaminated chronically with 137Cs did not demonstrate changes in the maximal response to carbachol, nor in the cholinergic sensitivity of rat jejunal epithelium. In conclusion, this study shows that chronic ingestion of 137Cs over 3 mo at postaccidental doses exerts few biological effects on the epithelium of rat jejunum with regard to morphology, inflammation status, apoptosis/proliferation processes, and secretory functions.

Morphological changes of cell proliferation and apoptosis in rat jejunal mucosa at different ages.

To study the changes of cell proliferation and apoptosis in rat jejunal epithelium at different ages. Cell proliferation and apoptosis of the jejunal mucosal and glandulous epithelia from birth to postnatal 12(th) month were observed using immunocytochemistry (ICC), and TUNEL method. The height of villus, the thickness of muscle layer and the number of goblet cells in jejunal mucosal and glandulous epithelia were measured by BeiHang analytic software and analyzed by STAT. (1) Proliferating cell nuclear antigen (PCNA) positive cells of jejunal glandulous recess were found and increased in number from birth to the postnatal 3(rd) month. The number of PCNA positive cells peaked in the postnatal 3(rd) month, and decreased from then on. (2) The number of apoptotic cells also peaked in the postnatal 3(rd) month, showing a similar trend to that of the PCNA positive cells. (3) The height of jejunal villus increased after birth, peaked in the postnatal 3(rd) month and decreased from then on. The jejunal muscle layer became thicker in the postnatal 3(rd) week and the postnatal 12(th) month. The number of goblet cells of the jejunal mucosal and glandulous epithelia had a linear correlation with age. (1) PCNA positive cells are distributed in the jejunal glandulous recess. (2) Apoptotic cell number peaks in the postnatal 3(rd) month, indicating that cell proliferation and apoptosis are developed with the formation of digestive metabolism as rat grows to maturity. (3) The thickness of jejunal muscle layer increases to a maximum in the postnatal 3(rd) week, which may be related to the change in diet from milk to solid food. (4) The number of goblet cells increases rapidly in the postnatal 3(rd) week, probably due to ingestion of solid food.

Uniform apicobasal polarity of microtubules and apical location of γ-tubulin in polarized intestinal epithelium in situ

Polarized differentiation of the intestinal epithelium has been previously shown to depend on an intact microtubular system that is essential for vectorial delivery of apical membrane proteins to the apical cell surfaces. Uniform alignment and polarization of microtubules have been suggested to provide the ultrastructural basis for vectorial transport between the Golgi apparatus and the apical cell surface. In the present study we applied the hook decoration technique to analyse the polarity of microtubules in the rat jejunal epithelium. By immunocytochemistry we studied the subcellular location of gamma-tubulin, an essential component of microtubule-organizing centers. Microtubules were found to be mainly aligned parallel to the apicobasal axis of the cells and to extend from the subterminal space underneath the apical terminal web down to the cellular basis. We found that 98% out of 1122 decorated microtubules displayed uniform apicobasal polarity with the growing ends (plus ends) pointing basally and the non-growing ends (minus ends) pointing towards the cellular apex. No differences were observed with respect to microtubular polarity between the apical, perinuclear and infranuclear cellular portions. Immunostaining specific for gamma-tubulin was restricted to the apical subterminal space underneath the rootlets of microvilli. These findings indicate that the apical subterminal space of enterocytes serves as the predominant if not exclusive microtubule-organizing compartment from which uniformly polarized microtubules grow out with their plus ends towards the cellular basis.

Cloning and functional expression of a cDNA from rat jejunal epithelium encoding a protein (4F2hc) with system y+L amino acid transport activity.

Two different protein families, designated CAT (cationic amino acid transporter) and BAT (broad-specificity amino acid transporter) mediate the plasma membrane transport of cationic amino acids in animal cells. CAT transporters have 12-14 transmembrane domains and are selective for cationic amino acids. BAT proteins, in contrast, have one to four transmembrane domains and induce the transport of both cationic and zwitterionic amino acids when expressed in Xenopus oocytes. Mutations in the human BAT gene cause type I cystinuria, a disease affecting the ability of intestinal and renal brush border membranes to transport cationic amino acids and cystine. We have used functional expression cloning in oocytes to isolate a BAT-related cDNA from rat jejunal epithelium. The cDNA encodes the rat 4F2 heavy chain (4F2hc) cell-surface antigen, a 527-residue (60 kDa) protein that is 26% identical in amino acid sequence with rat renal BAT (also known as NBAT/D2). Expression of rat jejunal 4F2hc in oocytes induced the lysine-inhibitable Na+-dependent influx of leucine and the leucine-inhibitable Na+-independent influx of lysine. Lysine efflux was stimulated by extracellular (Na+ plus leucine). These characteristics identify the expressed amino acid transport activity as system y+L, a transporter that has been implicated in basal membrane transport of cationic amino acids in intestine, kidney and placenta.

Functional and molecular characterization of NHE3 expression during ontogeny in rat jejunal epithelium.

Ontogenic changes occur in intestinal brush-border membrane vesicle (BBMV) Na+/H+ exchange activity. The present studies were designed to investigate ontogenic changes in Na+/H+ exchanger (NHE) isoform 3 in rat jejunum. pH-dependent Na+ uptake was assayed in four age groups of rats in the presence of 0, 50, or 800 microM HOE-694, a specific NHE inhibitor with differential sensitivities for NHE2 [inhibition constant (Ki) = 5 microM in PS120 fibroblasts] and NHE3 (Ki = 650 microM). Results showed that NHE2 and NHE3 contribute to basal BBMV uptake at all ages. Uptake levels were highest in 6-wk-old rats, lower in adult rats, and lowest in 2-wk-old (suckling) and 3-wk-old (weanling) rats, NHE3 contribution ranged from 92% at 6 wk of age to 59% at 2 and 3 wk of age. NHE3 inhibition by 800 microM HOE-694 was 38-45%. Statistical analysis showed that HOE-694 had a significant effect at both concentrations at all ages and that differences were present between all ages except 2- and 3-wk rats (at all HOE-694 concentrations). Northern blot analyses of jejunal mucosa showed lowest NHE3 mRNA levels in 2-wk animals and higher levels in all other age groups. Polyclonal antibodies were developed against an NHE3 COOH-terminal fusion protein, and antiserum was characterized with NHE3-transfected PS120 cells and by immunohistochemistry. Western blot analyses showed lowest protein levels in 2-wk animals and higher levels in the other ages. Suckling rats were subcutaneously injected with methylprednisone (MP) for 2 days and killed 1 day later. Northern blot analyses showed a twofold increase in NHE3 mRNA expression with MP treatment. Immunoblot analyses showed a 2.5-fold increase in NHE3 immunoreactive protein levels with MP injection. Overall, these data suggest that NHE3 is regulated during ontogeny and that ontogenic changes are most apparent around the time of weaning. Furthermore, the data suggest that NHE3 is regulated at transcriptional and posttranscriptional levels during mammalian intestinal development.

Molecular cloning and functional expression of cDNAs encoding a human Na+-nucleoside cotransporter (hCNT1).

We report identification of a new human nucleoside transporter protein by molecular cloning and functional expression of its cDNA. Previously, we used expression selection in Xenopus oocytes to isolate a cDNA from rat jejunal epithelium encoding the pyrimidine-selective Na+-dependent nucleoside transporter rCNT1 (Q.-Q. Huang, S. Y. M. Yao, M. W. L. Ritzel, A. R. P. Paterson, C. E. Cass, and J. D. Young. J. Biol. Chem. 269: 17757-17760, 1994). cDNAs for a human homologue of rCNT1, designated hCNT1, have been isolated from human kidney by hybridization cloning and reverse transcriptase polymerase chain reaction amplification strategies. hCNT1 was 83% identical to rCNT1 in amino acid sequence and exhibited the transport characteristics of an Na+-dependent nucleoside transporter with selectivity for pyrimidine nucleosides and adenosine when expressed in Xenopus oocytes. Deoxyadenosine, which undergoes net renal secretion, and guanosine were poor permeants. hCNT1 did, however, transport 3'-azido-3'-deoxythymidine. This is the first demonstration that members of the CNT family exist in human cells and provides evidence of their involvement in the renal transport of physiological nucleosides and nucleoside drugs. The hCNT1 gene was mapped to chromosome 15q25-26.

Cloning and functional expression of a complementary DNA encoding a mammalian nucleoside transport protein.

Expression screening in Xenopus oocytes was used to isolate a cDNA from rat jejunal epithelium encoding a Na(+)-dependent nucleoside transport protein (named cNT1). The cDNA sequence of cNT1 predicts a protein of 648 amino acids (relative molecular mass 71,000) with 14 potential transmembrane domains. Data base searches indicate significant sequence similarity to the NUPC proton/nucleoside symporter of Escherichia coli. There is no sequence similarity between cNT1 and proteins of mammalian origin. Functionally, cNT1 exhibited the transport characteristics of the nucleoside transport system cit (selective for pyrimidine nucleosides and adenosine) and accepted both 3'-azido-3'-deoxythymidine (AZT) and 2',3'-dideoxycytidine (ddC) as permeants (Km = 0.49 and 0.51 mM, respectively). The demonstration of transport of AZT by cNT1 expressed in Xenopus oocytes provides the first direct evidence that AZT enters cells by transporter-mediated processes, as well as by passive diffusion. Consistent with the tissue distribution of system cit transport activity, transcripts for cNT1 were detected in kidney as well as jejunum. cNT1 therefore belongs to a potential new gene family and may be involved in the intestinal absorption and renal handling of pyrimidine nucleoside analogs used to treat acquired immunodeficiency syndrome (AIDS).

Antisecretory activity of the alpha2-adrenoceptor agonist, xylazine in rat jejunal epithelium

Xylazine, an alpha 2 adrenoceptor agonist, reduces short circuit current (SCC) in epithelial preparations of rat jejunum. The alpha 2 antagonist yohimbine, abolished this response while prazosin was without effect. The cyclooxygenase inhibitor, piroxicam, also attenuated xylazine responses indicating that the antisecretory effects were dependent upon endogenous eicosanoid formation. If the secretory state of piroxicam treated tissues was restored by addition of either forskolin, vasoactive intestinal polypeptide (VIP), prostaglandin E2 (PGE2) or isobutyl-1-methyl-xanthine (IBMX) then subsequent additions of xylazine were effective in reducing SCC. All these agents are known to increase SCC and cause Cl secretion by elevating intracellular cAMP. In addition, xylazine was also able to inhibit the Ca2+-mediated secretory responses of carbachol (CCh) and substance P (SP) in rat jejunum. This ability of xylazine to inhibit cAMP- and Ca2+-mediated secretion may indicate that alpha 2 adrenoceptors interact with more than one type of G protein or alternatively suggests a more general interaction between second messenger systems within epithelia of the small intestine.

Regulation of intestinal epithelial cell growth by transforming growth factor type beta.

A nontransformed rat jejunal crypt cell line (IEC-6) expresses transforming growth factor type beta 1 (TGF-beta 1) mRNA, secretes latent 125I-labeled TGF-beta 1 competing activity into culture medium, and binds 125I-labeled TGF-beta 1 to specific, high-affinity (Kd = 3.7 pM) cell surface receptors. IEC-6 cell growth is markedly inhibited by TGF-beta 1 and TGF-beta 2 with half-maximal inhibition occurring between 0.1 and 1.0 ng of TGF-beta 1 per ml. TGF-beta 1-mediated growth inhibition is not associated with the appearance of biochemical markers of enterocyte differentiation such as alkaline phosphatase expression and sucrase activity. TGF-beta 1 (10 ng/ml) increases steady-state levels of its own mRNA expression within 8 hr of treatment of rapidly growing IEC-6 cells. In freshly isolated rat jejunal enterocytes that are sequentially eluted from the crypt villus axis, TGF-beta 1 mRNA expression is most abundant in terminally differentiated villus tip cells and least abundant in the less differentiated, mitotically active crypt cells. We conclude that TGF-beta 1 is an autoregulated growth inhibitor in IEC-6 cells that potentially functions in an autocrine manner. In the rat jejunal epithelium, TGF-beta 1 expression is most prominently localized to the villus tip--i.e., the region of the crypt villus unit that is characterized by the terminally differentiated phenotype. These data suggest that TGF-beta 1 may function in coordination of the rapid cell turnover typical for the intestinal epithelium.

Neuropeptide Y antagonises secretagogue evoked chloride transport in rat jejunal epithelium

Neuropeptide Y (NPY) inhibits electrogenic Cl secretion in rat jejunal epithelium under voltage clamp conditions. This effect is dependent upon endogenous eicosanoid formation since it is blocked by the cyclooxygenase inhibitor, piroxicam, which itself has an inhibitory action upon chloride secretion. A number of chloride secretagogues have been examined for their ability to restore the antisecretory effects of NPY. Data presented here shows that NPY responsiveness is restored, in piroxicam pretreated tissues, by vasoactive intestinal polypeptide (VIP), forskolin, prostaglandin E2 (PGE2) isobutyl-1-methyl-xanthine (IBMX) and dibutyryl cAMP added prior to the neuropeptide. While all these agents cause chloride secretion by elevating intracellular cAMP, NPY is also effective in inhibiting the secretory effects of carbachol (CCh) and substance P (SP), agents believed to act by raising intracellular calcium (Cai). Although there is evidence that NPY can inhibit adenylate cyclase, its ability to attenuate chloride secretion brought about by secretagogues acting through both adenylate cyclase and calcium mechanisms, implies that NPY has either a more general fundamental mechanism or has multiple interactions with different second messenger systems.

Phosphate transport across rat jejunum: influence of sodium, pH, and 1,25-dihydroxyvitamin D3

Inorganic phosphate (Pi) transport in intact, rat jejunal epithelium was measured in vitro under short-circuited conditions. Transepithelial net absorptive Pi flux increased linearly with increases in extracellular sodium concentration ([Na]) up to 144 mM. Transmucosal border Pi influx, in contrast, displayed a biphasic Na dependency. Pi influx increased as [Na] was raised from 0 to 100 mM. A further increase in [Na] to 144 mM caused unanticipated reduction in Pi influx. The reason for this dissociation between transmucosal border influx and transepithelial absorptive flux is not clear. We then examined the effect of changes in extracellular pH on Pi influx. Reduction in pH from 7.4 to 6.0 was associated with 150% increase in Pi influx, an observation consistent with the reported reciprocal relation between intestinal brush-border membrane vesicles (BBMV) Pi uptake and extravesicular pH. In contrast to BBMV data, however, a smaller increase (50%) in mucosal Pi influx was noted in intact epithelium when pH was increased from 7.4 to 8.5. Under optimized conditions for Pi influx, i.e., [Na] = 90 mM and pH 6.0, the effect of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] on Pi influx was measured. The total influx could be resolved into a saturable, Na-dependent and a nonsaturable, Na-independent component. 1,25(OH)2D3 stimulated the saturable component of Pi influx.

Interaction of Peptide YY with Rat Intestinal Epithelial Plasma Membranes: Binding of the Radioiodinated Peptide

High affinity binding sites for peptide YY (PYY) have been identified and characterized in plasma membranes prepared from rat jejunal epithelium by studying the kinetics, stoichiometry, and chemical specificity of the interaction of 125I-labeled PYY with membranes. Binding of [125I]PYY was rapid, saturable, reversible, specific, and depended on temperature, pH, and ionic strength. In optimized steady state conditions of binding (2 h of incubation at 15 C), the degradation of both [125I] PYY and binding sites did not exceed 20%. The concentration dependence of PYY binding, determined by adding increasing concentrations of [125I]PYY, indicated that specific binding saturated at 2-3 nM peptide. Scatchard analysis revealed a single class of binding sites with a dissociation constant (Kd) of 434 +/- (SE) 56 pM and a binding capacity of 336 +/- 41 fmol/mg protein (n = 11). Identical results were obtained when increasing concentrations of unlabeled PYY were added to a fixed concentration of [125I]PYY, indicating that the radioiodinated peptide has the same apparent affinity as native PYY. Peptides structurally unrelated to PYY, such as members of the vasoactive intestinal peptide family, insulin, or cholecystokinin octapeptide, were unable to compete with [125I]PYY for binding to membranes. Rat, human, and avian pancreatic polypeptides, which display, respectively, 42%, 47%, and 53% homology with PYY, did inhibit [125I]PYY binding but with an approximate or equal to 100,000-fold lower potency than PYY, indicating the strict structural requirement for recognition by PYY binding sites. In contrast, natural or synthetic neuropeptide Y, which has 25 out of 36 amino acids in common with PYY, retained a high affinity for PYY binding sites [only 4.7 +/- 1.2 (n = 5) times lower than that of PYY]. Specific [125I]PYY binding was particularly high in the upper small intestine and could not be detected in stomach, large intestine, or liver. These findings indicate that rat small intestinal epithelium expresses specific binding sites for the candidate gut hormone PYY that also binds the neuropeptide Y with high affinity, suggesting that the two peptides may regulate the function of small intestinal epithelium, through interaction with a common receptor site.

Ultrastructural studies on the secretory mechanism of goblet cells in the rat jejunal epithelium.

Goblet cells in the jejunal epithelium of adult and suckling rats were studied with transmission electron microscope. The use of triple fixation, i.e. osmium-aldehyde-osmium has some advantages in the preservation of mucous substances and some cell organelles. Cytochemical demonstration of polysaccharide and glycoprotein by methenamine silver as well as detection of TPPase and acid phosphatase (AcPase) were performed. Accumulation of secretory substances and combination with sugar moieties occurred in the internal sacs of Golgi lamellae, where TPPase activity was recognized. No rigid lamella usually positive to AcPase was found in goblet cells, though it was easily found in the columnar absorptive cells. AcPase was positive in some small immature mucus droplets as well as lysosomes. As the differentiation of goblet cells advances, mucus droplets come into contact and fuse to each other. As a result of the close contact of two adjacent membranes of mucus droplets, a pentalaminar structure is first formed. Then the central dense lamina disappears and the membrane becomes trilaminar. Disappearance of one of the apposed unit membranes, leaving another unit membrane, may take place. Finally the single unit membrane left between the adjacent droplets is also ruptured and complete fusion occurs. The mechanism of extrusion of mucous substance is complicated. Some mucus droplets located at the most superficial area may be released by the mechanisms of exocytosis. More frequently, however, deeply located droplets fuse to each other and a huge vacuole is made in the apical cytoplasm. The substance filling the vacuole contains debris of cytoplasmic matrix and droplet membranes along with mucous secretory substance. These substances derived from different sources are expelled into the lumen at the same time. This may be classified into the apocrine mechanism, because a part of the cytoplasm and membrane are lost. The typical apocrine secretion, i.e. pinching off of cytoplasmic projection containing mucus droplets, was also found in the goblet cells. Thus, two or three different mechanisms of secretion discharge may possibly occur simultaneously in goblet cells.

Transition between columnar absorptive cells and goblet cells in the rat jejunal epithelium.

Electron microscopic observation of the jejunal epithelium of rats demonstrated morphological evidence of a transition between columnar absorptive cells and growing goblet cells. The columnar cells in both the villi and crypts have features suggestive of absorptive functions. They are provided with apical invaginations continuous to the intermicrovillous space. Absorbed lipid is observed in small vesicles in the terminal web layer, and chylomicrons derived here from are contained in large vacuoles near the Golgi apparatus. Ferritin particles artificially infused into the gut lumen were absorbed into the vacuoles in the subapical zone of columnar cells of suckling rats. Growing goblet cells situated in the crypt epithelium contain surface invaginations and lysosomes which are the same in structure as those found in absorptive cells nearby. Fat droplets evidently absorbed by the growing goblet cell were observed among immature mucus droplets. Artificially infused ferritin particles were found in vacuoles and lysosomes near the Golgi apparatus of some goblet cells of suckling rats. Some goblet cells on the intestinal villi of suckling rats looked immature and their microvilli and cytoplasmic matrix were clear like those of columnar absorptive cells. The transition between these goblet cells with clear cytoplasm and the mature goblet cells with dark cytoplasm was observed. These morphological evidences indicate that some of columnar cells already differentiated to absorptive cells are capable of transforming into mucus-producing (goblet) cells. It is suggested that not only undifferentiated columnar cells in the crypt base but also considerably differentiated columnar cells with absorptive function can differentiate into goblet cells.

Characteristics of rat jejunal transport of tryptophan

The parameters of rat jejunal transport of tryptophan have been examined. The interactions between tryptophan and lysine or methionine have been reexamined, and some aspects of the trans effects of cellularly accumulated amino acids have been studied. It has been demonstrated that: 1. (1) The influx of tryptophan across the jejunal brush border ( J mc Trp ) can be accounted for by the carrier of α-aminomonocarboxylic acids alone. 2. (2) Tryptophan competes with lysine for the carrier of basic amino acids across the brush border membrane without itself being transported by this carrier. 3. (3) Lysine has neither cis nor trans effects on J mc Trp , whereas intracellular tryptophan is highly inhibitory to J mc Lys . 4. (4) The intracellular concentration of lysine and of tryptophan, [Lys] c and [Trp] c, are unaffected by tryptophan and lysine, respectively, although the transmural fluxes, from the mucosal side to the serosal side, J ms , of lysine, J ms Lys , and of tryptophan, J ms Trp , are inhibited by tryptophan and lysine, respectively. The latter effects thus represent inhibitory interactions at the basolateral membrane. 5. (5) Methionine is a potent cis and trans inhibitor of J mc Trp , but stimulates J ms Trp and reduces [Trp] c. 6. (6) Methionine causes trans acceleration of the influx of lysine across the brush border membrane, J mc Lys , but has no effect on the influx of galactose, J mc Gal . 7. (7) Leucine causes trans inhibition of J mc Leu . 8. (8) Tryptophan does not cause cis inhibition of J mc staggered Gal , but is a strong trans inhibitor of J mc Gal . 9. (9) Cellularly accumulated tryptophan appears to accelerate the eventual decline in transepithelial potential difference and short-circuit current. These results are consistent with the conclusions that: 1. (1) Tryptophan is transported across the brush border membrane by the carrier of neutral amino acids alone, but leaves the cell across the basolateral membrane by a mechanism used by lysine also. 2. (2) Leucine, methionine and probably tryptophan have a transeffect on the transport of neutral amino acids across the brush border membrane which may represent a phenomenon which can appropriately be termed decelerating exchange diffusion. 3. (3) Cellularly accumulated tryptophan has a strong and indiscriminate depressive effect on all transport functions of rat jejunal epithelium.