Leptin reduces glucose transport and cellular ATP levels in INS-1 beta-cells.

Leptin suppresses insulin secretion by opening ATP-sensitive K(+) (K(ATP)) channels and hyperpolarizing beta-cells. We measured the intracellular concentration of ATP ([ATP](i)) in tumor-derived beta-cells, INS-1, and found that leptin reduced [ATP](i) by approximately 30%, suggesting that the opening of K(ATP) channels by leptin is mediated by decreased [ATP](i). A reduction in glucose availability for metabolism may explain the decreased [ATP](i), since leptin (30 min) reduced glucose transport into INS-1 cells by approximately 35%, compared to vehicle-treated cells. The twofold induction of GLUT2 phosphorylation by GLP-1, an insulin secretagogue, was abolished by leptin. Therefore, the acute effect of leptin could involve covalent modification of GLUT2. These findings suggest that leptin may inhibit insulin secretion by reducing [ATP](i) as a result of reduced glucose availability for the metabolic pathway. In addition, leptin reduced glucose transport by 35% in isolated rat hepatocytes that also express GLUT2, suggesting that glucose transport may also be altered by leptin in other glucose-responsive tissues such as the liver.

Glucose transporter gene expression in lactating bovine gastrointestinal tract.

Absorption of glucose from the intestine and probably from various portions of the gastrointestinal tract (GIT) in lactating dairy cows can have profound implications because the availability of glucose, or glucose precursors, seems to be a limiting factor for milk synthesis. This article reports on the expression of the Na+/glucose cotransporter gene (SGLT1) in mucosal tissues from rumen, omasum, abomasum, duodenum, jejunum, ileum, and cecum of lactating cows. Sodium/glucose cotransporter mRNA transcripts were detected in polyadenylated RNA of all the GIT tissues examined and in total RNA from rumen, omasum, duodenum, jejunum, ileum, and cecum. The pattern of SGLT1 mRNA distribution in these tissues was different from that of the facilitative glucose transporter GLUT5. Sodium/glucose cotransporter protein was also detected in small intestinal membranes of lactating dairy cows. Sodium-dependent glucose uptake across small intestinal brush border membrane vesicles could be resolved into linear nonsaturable (diffusion) and saturable, carrier-mediated components. The diffusion constant of glucose from jejunal membrane vesicles was 18.3+/-2.1 pmol x mM(-1) x mg protein(-1) x s(-1). The saturable component of glucose uptake had a Vmax of 27.1+/-5.8 pmol x mg protein(-1) x s(-1) and a Km of .12+/-.05 mM. Similar transport kinetics were found in the duodenum, and they were less in the ileum. These data suggest that the GIT of lactating cows is capable of active transport of glucose from the lumen across the brush border membrane of the epithelial cells. In addition, the relatively high abundance of SGLT1 mRNA in the rumen, omasum, and cecum may imply new functional and nutritional roles for these tissues in the active transport of glucose.

Basolateral D-glucose transport activity along the crypt-villus axis in rat jejunum and upregulation induced by gastric inhibitory peptide and glucagon-like peptide-2.

Phloridzin-insensitive D-glucose uptake into enterocytes isolated sequentially from along the crypt-villus axis showed the majority of transport activity to reside in cells from the upper third of the villus. In contrast, total postnuclear glucose transporter 2 (GLUT2) protein content of the cells was high even close to the crypt and was almost constant for the upper 80% of the villi. A 4 h lumenal perfusion in vivo with 100 mM D-glucose prior to harvesting the enterocytes produced a 2- to 3-fold increase in phloridzin-insensitive D-glucose uptake which extended down 70% of the villus. Vascular infusion in vivo with either 800 pM gastric inhibitory polypeptide (GIP) or glucagon-like peptide-2 (GLP-2) prior to harvesting enterocytes produced the same response as lumenal glucose, while glucagon like peptide-1 (GLP-1) had no effect. Inclusion of 30 microM brefeldin A (BFA), an inhibitor of protein trafficking, in the lumenal perfusate produced a small, but not significant, increase in the control uptake profile along the villus in isolated enterocytes. However, BFA significantly reduced the upregulation induced by lumenal glucose and vascular GIP and blocked the stimulation produced by vascular GLP-2. Biotinylation of surface proteins and isolation with protein A indicated that there was no change in the membrane abundance of GLUT2 after GLP-2 treatment. These results are discussed in relation to the role of gastrointestinal peptide hormones in controlling intestinal hexose transport and the possibility of protein trafficking being involved in mediating the upregulation of GLUT2 activity in the enterocyte basolateral membrane.

Cholecystokinin decreases intestinal hexose absorption by a parallel reduction in SGLT1 abundance in the brush-border membrane.

The dual lumenaly and vascularly perfused small intestine was used to determine the mechanism by which cholecystokinin octapeptide (CCK-8) decreases the rate of glucose absorption. With CCK-8 in the vascular perfusate the rate of 3-O-methyl-D-glucose absorption decreased, whereas the rate of D-fructose absorption was unaffected. The substrate pool size within the tissue during steady-state transport, in the presence and absence of CCK-8, was estimated by compartmental analysis of the 3-O-methyl-D-glucose washout into the vascular bed. When CCK-8 was included in the vascular perfusate, the absorptive cell pool size decreased when compared with untreated tissue. Both the steady-state hexose absorption data and the washout studies indicated that the locus of action of CCK-8 was the SGLT1 transporter located in the brush-border membrane. The SGLT1 protein abundance in isolated brush-border membranes, as quantified by Western blotting, showed a decrease that paralleled the decrease in the steady-state transport rate induced by CCK-8. These results indicate that CCK-8 diminishes the rate of intestinal hexose absorption by decreasing SGLT1 protein abundance in the brush-border membrane of the rat jejunum and therefore provides evidence for acute enteric hormonal regulation of the rate of glucose absorption across the small intestine.

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.

GLUT-1 mediation of rapid glucose transport in dolphin (Tursiops truncatus) red blood cells.

D-Glucose entry into erythrocytes from adult dolphins (Tursiops truncatus) was rapid, showed saturation at high substrate concentrations, and demonstrated a marked stimulation by intracellular D-glucose. Kinetic parameters were estimated from the concentration dependence of initial rates of tracer entry at 6 degrees C: for zero-trans entry, Michaelis constant (K(m)) was 0.78 +/- 0.10 mM and maximal velocity (Vmax) was 300 +/- 9 mumol.l cell water-1.min-1; for equilibrium exchange entry, K(m) was 17.5 +/- 0.6 mM and Vmax was 8,675 +/- 96 mumol.l cell water-1.min-1. Glucose entry was inhibited by cytochalasin B, and mass law analysis of reversible, D-glucose-displaceable, cytochalasin B binding gave values of 0.37 +/- 0.03 nmol/mg membrane protein for maximal binding and 0.48 +/- 0.10 microM for the dissociation constant. Dolphin glucose transporter polypeptides were identified on sodium-dodecyl sulfate-polyacrylamide gel electrophoresis immunoblots [using antibodies that recognized human glucose transporter isoform (GLUT-1)] as two molecular species, apparent relative molecular weights of 53,000 and 47,000. Identity of these polypeptides was confirmed by D-glucose-sensitive photolabeling of membranes with [3H]cytochalasin B. Digestion of both dolphin and human red blood cell membranes with glycopeptidase F led to the generation of a sharp band of relative molecular weight 46,000 derived from GLUT-1. Trypsin treatment of human and dolphin erythrocyte membranes generated fragmentation patterns consistent with similar polypeptide structures for GLUT-1 in human and dolphin red blood cells.

Kinetic analysis of L-glutamine transport into porcine jejunal enterocyte brush-border membrane vesicles.

L-Glutamine transport into porcine jejunal enterocyte brush border membrane vesicles was studied. Uptake was mediated by a Na(+)-dependent and a Na(+)-independent pathway as well as by diffusion. The initial rates of glutamine uptake over a range of concentrations is both Na(+)-gradient and Na(+)-free conditions were analyzed and kinetic parameters were obtained. Na(+)-dependent glutamine transport had a K(m) of 0.77 +/- 0.16 mM and a Jmax of 70.7 +/- 5.8 pmol mg protein-1 s-1; Na(+)-independent glutamine transport had a K(m) of 3.55 +/- 0.78 mM and a Jmax of 55.1 +/- 6.6 pmol mg protein-1 s-1. The non-saturable component measured with HgCl2-poisoned brush border membrane vesicles in the Na(+)-free condition contained passive diffusion and non-specific membrane binding and was defined to be apparent glutamine diffusion and the glutamine permeability coefficient (Kdiff) was estimated to be Kdiff = 3.78 +/- 0.06 pmol 1 mg protein-1 mmol-1 s-1. Results of inhibition experiments showed that Na(+)-dependent glutamine uptake occurred primarily through the brush border system-B degree transporters, whereas Na(+)-independent glutamine uptake occurred via the system-L transporters. Furthermore, the kinetics of L-leucine and L-cysteine inhibition of L-glutamine uptake demonstrated that neutral amino acids sharing the same brush border transporters can effectively inhibit each other in their transport.

Evidence that the transport-related proteins BAT and 4F2hc are not specific for amino acids: induction of Na+-dependent uridine and pyruvate transport activity by recombinant BAT and 4F2hc expressed in Xenopus oocytes.

Members of the BAT and 4F2hc gene family have one or, in the case of BAT, up to four transmembane domains and induce amino acid transport systems b(o,+) (BAT) and y+L (4F2hc) when expressed in Xenopus oocytes. System b(o,+) is a Na+-independent process with a broad tolerance for cationic and zwitterionic amino acids, whereas y+L exhibits Na+-independent transport of cationic amino acids (e.g., lysine) and Na+-dependent transport of zwitterionic amino acids (e.g., leucine). Mutations in the human BAT gene are associated with type I cystinuria, a genetic disease affecting the ability of intestinal and renal brush border membranes to transport cationic amino acids and cystine. An unresolved question is whether BAT and 4F2hc themselves have catalytic (i.e., transporting) activity or whether they operate as activators of other, as yet unidentified, transporter proteins. In this report, we have investigated the transport of representatives of four different classes of organic substrates in Xenopus oocytes following injection with rat BAT or 4F2hc RNA transcripts: leucine (a control amino acid substrate), uridine (a nucleoside), pyruvate (a monocarboxylate), and choline (an amine). Both recombinant proteins induced small, statistically significant Na+-dependent fluxes of uridine and pyruvate but had no effect on choline uptake. In contrast, control oocytes injected with transcripts for conventional nucleoside and cationic amino acid transporters (rat CNT1 and murine CAT1, respectively) showed no induction of transport of either leucine or pyruvate (CNT1) or uridine or pyruvate (CAT1). These findings support the idea that BAT and 4F2hc are transport activators and minimize the possibility that they have intrinsic transport capability. The transport-regulating functions of these proteins may extend to permeants other than amino acids.

Inhibition of glucose absorption in the rat jejunum: a novel action of alpha-D-glucosidase inhibitors.

BACKGROUND & AIMS: alpha-D-Glucosidase inhibitors act primarily by decreasing disaccharide hydrolysis and thus reduce the amount of free monosaccharides available for absorption. A novel action of alpha-D-glucosidase inhibitors is presented, indicating a direct effect on free glucose absorption by the rat jejunum. METHODS: The jejunum was isolated and free hexose was measured using in vivo single-pass luminal perfusion and dual vascular and luminal single-pass in vitro perfusion. Xenopus oocytes were injected with RNA transcript encoding recombinant sodium-glucose cotransporter 1, and uptake of 3H-labeled 3-O-methyl-D-glucopyranose (3-O-MG) was assessed. RESULTS: Acarbose (0.1 mg/mL), added to the lumen, decreased D-glucose absorption by 20% in vivo. Addition of 0.1 or 1.0 mg/mL acarbose to the lumen in vitro decreased the appearance of 3-O-MG in the vascular effluent by 28% and 60%, respectively. Accumulation of D-glucose within the enterocytes was decreased significantly by 67% and 79% when acarbose (1 mg/mL) or phloridzin (2 mmol/L), respectively, were present in the luminal perfusate. In contrast, acarbose did not affect the transport rate of free D-fructose and did not inhibit 3-O-MG uptake in oocytes expressing sodium-glucose cotransporter 1. CONCLUSIONS: The findings indicate that alpha-D-glucosidase inhibitors act specifically on the entry of free glucose into the enterocyte, an additional means by which they can reduce postprandial hyperglycemia.

Nucleoside transport in erythrocytes from bottle-nosed dolphin (Tursiops truncatus).

Entry of adenosine, and thymidine, into erythrocytes from adult dolphins was rapid, showed saturation at higher substrate concentrations, and was strongly inhibited by low concentrations of nitrobenzylthioinosine (NBMPR). Kinetic parameters were estimated from the concentration dependence of initial rates of tracer entry at 21 degrees C, as K(m) 0.14 +/- 0.05 mM and Vmax 24.4 +/- 1.9 mumol/litre cell water/sec for zero trans entry of adenosine, and K(m) 0.96 +/- 0.21 mM and Vmax 25.4 +/- 1.7 mumol/litre cell water/sec for thymidine. Adenosine, and thymidine, entry were inhibited by both purine and pyrimidine nucleosides. Mass law analysis of a saturable component of nitrobenzylthioinosine binding to dolphin red cell membranes gave values of Bmax 65.4 +/- 1.2 pmol/mg protein, and K(d) of 1.53 +/- 0.08 nM for a single class of sites. Photo-irradiation of dolphin red cell membranes in the presence of tritiated nitrobenzylthioinosine led to radioactive labeling of polypeptides M(r) 52, 500-58,000, on SDS-PAGE.

Upregulation of SGLT-1 transport activity in rat jejunum induced by GLP-2 infusion in vivo.

The effect of in vivo infusion of the peptide hormone glucagon-like peptide 2 (GLP-2) on glucose transport across the rat jejunal brush-border membrane (BBM) was assessed using isolated membrane vesicles. A 2-h infusion of GLP-2 produced a marked acceleration of sodium-dependent glucose uptake into BBM vesicles with a significant overshoot. There was no change in vesicle space or permeability resulting from the hormone infusion. Kinetic analysis showed this stimulation to be the result of a three-fold increase in the maximal rate of transport, with no consistent change in the affinity constant (Km). The time course of this response showed that the effect was observable, but smaller, after only 30 min of hormone infusion and was maximal after 1 h. Sodium-dependent phloridzin binding to the membrane vesicles showed a parallel increase in maximal binding after 1 and 2 h of hormone infusion. Western blotting showed a similar increase in sodium-dependent glucose transporter 1 (SGLT-1) abundance. The effect of GLP-2 could be blocked by luminal brefeldin A or wortmannin. These results indicate that GLP-2 is able to induce trafficking of SGLT-1 from an intracellular pool into the BBM within 60 min and that phosphoinositol 3-kinase may well be involved in the intracellular signaling pathway in this response.

Effect of cholecystokinin and related peptides on jejunal transepithelial hexose transport in the Sprague-Dawley rat.

An in situ dual vascular and luminal perfusion technique was used to study the effect of cholecystokinin octapeptide (CCK-8) on the transport of hexoses by the jejunum of the Sprague-Dawley rat from the lumen to the vascular bed. The lumen of the jejunum was perfused with hexoses in oxygenated Krebs buffer, while the superior mesenteric artery was infused with Krebs buffer containing Ficoll 70 as a plasma expander. CCK-8 (0.8-8 pM) in the vascular infusate selectively reduced hexose transport in a dose-dependent manner by 20-47%, although having no effect on L-glucose or L-leucine absorption. Vascular tetrodotoxin did not block CCK-8 inhibition, whereas a specific CCK-A receptor antagonist, lorglumide, did. The CCK-B receptor agonist cholecystokinin tetrapeptide had a small effect on hexose absorption, whereas somatostatin-14 and -28 had no effect. These results suggest that cholecystokinin can decrease intestinal absorption of hexoses in the small intestine, acting via CCK-A-type receptors.

The effect of GIP and glucagon-like peptides on intestinal basolateral membrane hexose transport.

The effect of gastric inhibitory polypeptide (GIP) and the related glucagon-like peptides-1 and -2 (GLP-1 and GLP-2) on jejunal basolateral membrane glucose transport was investigated to determine if the upregulation produced by luminal hexoses could be explained by the release of one or more of these peptides. Luminal perfusion of the rat jejunum for 4 h, under pentobarbital sodium anesthesia, with 100 mM D-glucose produced a significant increase in plasma GIP levels. Vascular infusion of saline containing 100-800 pM GIP also increased the maximal transport rate for carrier-mediated glucose uptake in jejunal basolateral membrane vesicles. The effect of vascular 400 pM GIP was maximal after 1 h and maintained out to 4 h. The effect of luminal glucose could be blocked by preinjection with anti-GIP antibodies, whereas an antineurotensin antibody had no effect. Vascular infusion with 800 pM GLP-1-(7-36) amide had no effect, but GLP-2 (400 and 800 pM) increased the D-glucose maximal transport rate. An anti-GLP antibody was able to block the response to luminal glucose.

Effect of luminal epidermal growth factor on enterocyte glucose and proline transport.

The effect of luminal epidermal growth factor (EGF; 60 ng/ml) and tyrphostin-51 (TYR; 10 microM), a tyrosine kinase inhibitor, on rabbit jejunal brush-border and basolateral membrane transport was investigated. In separate experiments, the effect of EGF, EGF and TYR, or TYR alone was examined in in vivo loops. In addition, Na+ permeability in brush-border membrane vesicles (BBMV) and the effect of Ca2+ channel blockade on EGF-stimulated glucose uptake were examined. Luminal EGF significantly (P < 0.0001) increased the maximal rate of transport (Vmax) for glucose and proline uptake in BBMV. TYR and Ca2+ channel blockade completely abolished the EGF-induced increase in glucose transport and in the case of TYR resulted in a significant reduction in Vmax compared with controls (P < 0.0001). The Michaelis-Menten constant did not differ in any experimental group. EGF had no effect on brush-border Na+ permeability or basolateral membrane glucose transport. The findings indicate a role for EGF in the acute regulation of jejunal brush-border membrane nutrient uptake. Furthermore, tyrosine kinase activity appears to be involved both in mediating EGF-induced alterations in transport function and in the maintenance of basal brush-border membrane function.

Intestinal morphology and transport after ileal resection in rat is modified by dietary fatty acids.

The authors tested the hypothesis that the intestinal morphology and uptake of nutrients after resection of the distal half of the small intestine of rats responds to alterations in the dietary content of saturated (SFA) and polyunsaturated (PUFA) fatty acids. Adult female Sprague-Dawley rats were subjected to a sham operation or to the surgical resection of the distal half of the small intestine, leaving the ileocecal valve intact. The animals were fed chow for 3 weeks, then either chow or isocaloric semisynthetic SFA or PUFA diets for a further 2 weeks. Food consumption, weight gain and jejunal mucosal surface area were unchanged after ileal resection. A microdensitometric autoradiographic technique was used to examine the distribution of 3H-leucine and 3H-lysine along the villus: approximately 70% of uptake occurred in the upper 30% of the enterocytes of the villus in chow-fed rats, and this portion was unchanged by ileal resection. The jejunal uptake of 40 mM of glucose, observed in vitro, was twice as high in animals that had undergone resection and were fed SFA than in those fed PUFA. In summary, (1) there is a separation between the adaptation of intestinal transport function and dynamic/static morphology after ileal resection, and (2) glucose uptake after ileal resection is enhanced by SFA in the diet and is not explained by any changes in the animals' food intake, weight gain or intestinal morphology.

GLUT-3 (brain-type) glucose transporter polypeptides in human blood platelets.

Antiserum directed against a C-terminal peptide of the human GLUT-3 (brain type) equilibrative glucose transporter isoform reacted with polypeptides M(r) 46,000-48,000 on immunoblots of human platelets. Photoirradiation of human platelets in the presence of 3H-cytochalasin B led to radiolabeling of polypeptides of identical mobility. This labeling was substantially reduced by preincubation the cells with 440 mM D-glucose, but not 440 mM L-glucose, consistent with glucose transporter function. Only traces of GLUT-1 (erythrocyte type) glucose transporter polypeptides were detected, and there was no evidence of GLUT-2 (liver type) transporter on immunoblots of platelet proteins.

Poly(A)+ RNA from the mucosa of rat jejunum induces novel Na(+)-dependent and Na(+)-independent leucine transport activities in in oocytes of Xenopus laevis.

Complementary DNA clones have been isolated recently from rat (D2) and rabbit kidney (rBAT) which induce increased Na(+)-independent Leu and Lys transport activities (System b0, +) when expressed in oocytes of Xenopus laevis. These cDNAs encode type II membrane glycoproteins which show significant homology to the heavy chain of the human and mouse 4F2 surface antigen (4F2hc). Injection of human 4F2hc cRNA into oocytes also results in induction of Leu/Lys transport activity, but with differing cation requirements for the two amino acids (Na(+)-dependent for Leu, Na(+)-independent for Lys: system y+L). System y+L is a newly discovered zwitterionic/cationic amino acid transporter first described in human erythrocytes. Here we have examined the characteristics of Leu transport in Xenopus oocytes microinjected with mRNA from the mucosa of rat jejunum. L-Leu uptake during 10 min (0.2 mM, 20 degrees C) reached 20 pmol/oocyte compared with endogenous uptake by water-injected oocytes of typically 3-4 pmol/oocyte. The expressed transport activity was 80% Na(+)-dependent. The Na(+)-dependent component of the expressed flux was saturable (Km app 0.20 mM) and inhibited by Lys, but not by Ala or Phe. The minor Na(+)-independent component of expressed Leu transport activity was also saturable (Km app 0.10 mM). Amino acid inhibition studies resolved this flux into two main components, one of which was inhibited by Lys, Ala and Phe and another which was only inhibited by Lys. There was a small residual component of Na(+)-independent Leu transport which was insensitive to inhibition by Lys. Experiments utilizing polymerase chain reaction (PCR) demonstrated the presence of both D2 and 4F2hc message in rat jejunum. Hybrid-depletion of jejunal mRNA with an antisense oligonucleotide complementary to D2 had no effect on the expression of Na(+)-linked Leu transport activity, but reduced the smaller Na(+)-independent component of Leu transport by 40%, suggesting only a minor role of D2 in the expression of rat intestinal Leu transport activity. Although the properties of Na(+)-dependent Leu transport were, with the exception of a lack of inhibition by Ala and Phe, consistent with erythrocyte y+L, hybrid-depletion of jejunal mRNA with an antisense oligonucleotide complementary to 4F2hc had no detectable effect on the expressed transport activity. We conclude, therefore, that mRNA from rat jejunum encodes novel Na(+)-dependent and Na(+)-independent transport activities unrelated to the D2/4F2hc glycoproteins.(ABSTRACT TRUNCATED AT 400 WORDS)

Crypt cell production rate, enterocyte turnover time and appearance of transport along the jejunal villus of the rat.

Intestinal nutrient absorption is subject to adaptation with, for example, diabetes, diet lipid variations (isocaloric semisynthetic diets enriched with saturated (S) or polyunsaturated (P) fatty acids), ileal resection and abdominal irradiation. These models were used in rats to assess dynamic morphology and distribution of amino acid transporter along the villus. The enterocyte migration rate (EMR) was measured using [3H]thymidine; the vincristine metaphase arrest technique was used to determine the crypt cell production rate (CCPR); quantitative autoradiography was used to assess the time and age of enterocytes when the uptake of 1 and 20 mM [3H]leucine and [3H]lysine was initiated along the villus. The enhanced jejunal uptake of nutrients which occurs after a 50% distal enterectomy was associated with a fall in EMR and CCPR, yet the enhanced nutrient uptake which occurs in diabetes is not associated with any alteration in EMR, CCPR, enterocyte transport pool (ETP), i.e., the length of the enterocyte column along with the villus containing amino acid transporter) or expression of transporter along the villus. The reduced uptake of nutrients in rats fed P as compared with S was associated with increased rather than decreased ETP and age of the enterocytes at the tip of the villus. The reduced nutrient uptake which occurs 3 days after abdominal irradiation was associated with increased EMR and CCPR, and reduced ETP and age of enterocytes of the tip of the villus. However, 14 days after irradiation when nutrient transport remains reduced, these parameters have returned to normal. Thus, alterations in nutrient transport may be associated with changes in the dynamic morphology of the intestine, but the two processes are not necessarily interdependent. We speculate that the changes in the dynamic morphology of the intestine, and the changes of amino acid transport which occurs in these models of intestinal adaptation, are independently controlled.

Evidence for a lactate-anion exchanger in the rat jejunal basolateral membrane.

BACKGROUND/AIMS: The mechanism by which lactate, absorbed from the intestinal lumen or generated within the epithelium, crosses the basolateral membrane of the enterocyte and enters the bloodstream has not previously been characterized in detail. METHODS: L-lactate uptake into and efflux from isolated jejunal basolateral membrane vesicles was investigated at room temperature using rapid filtration techniques. RESULTS: Furosemide sensitive uptake of L-lactate was unaffected by cis sodium or proton gradients but could be stimulated by a trans gradient of bicarbonate and chloride. Kinetic analysis showed uptake to consist of a saturable component with a Michaelis constant (Km) of 3.2 mmol/L and a maximum velocity (Vmax) of 67 pmol.mg protein-1 x s-1 and a nonsaturable alpha-4-hydroxy-cinnamic acid insensitive component. Pyruvate, butyrate, acetate, valerate, and propionate competitively inhibited lactate uptake into the vesicles. Efflux of lactate from preloaded vesicles was furosemide sensitive and accelerated by a trans bicarbonate gradient as well as by 10 mmol/L acetate, butyrate, and pyruvate. CONCLUSIONS: It is concluded that there is a short chain-fatty acid carrier system in the intestinal basolateral membrane, which operates as an anion exchanger.