Increased expression of ileal apical sodium-dependent bile acid transporter in postpartum rats (original) (raw)
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Journal of Clinical Investigation, 1995
Sodium-dependent bile acid transport in the rat ileum is abruptly expressed at weaning. Degenerate oligonucleotides, based on amino acid sequence identities between the rat liver and hamster ileal transporters, were used to amplify a rat ileal probe. A 1.2-kb cDNA clone, which contains the full coding region (348 amino acids, 38 kD), was isolated by hybridization screening. In vitro translation yielded a 38-kD protein which glycosylated to 48 kD. Sodium-dependent uptake of taurocholate was observed in oocytes injected with cRNA. Northern blot analysis revealed a 5.0-kb mRNA in ileum, kidney, and cecum. A 48-kD protein was detected in ileal brush border membranes and localized to the apical border of villus ileal enterocytes. mRNA and protein expression, which were negligible before weaning, increased dramatically at weaning. Nuclear transcription rates for the transporter increased 15-fold between postnatal days 7 and 28. The apparent molecular weight of the transporter also increased between days 19 and 28. In summary, the developmental regulation of the rat ileal sodium-dependent bile acid cotransporter is characterized by transcriptionally regulated increases in mRNA and protein levels at the time of weaning with changes in apparent molecular weight of the protein after weaning.
Journal of Clinical Investigation, 1997
Although bile acid transport by bile duct epithelial cells, or cholangiocytes, has been postulated, the details of this process remain unclear. Thus, we performed transport studies with [ 3 H]taurocholate in confluent polarized monolayers of normal rat cholangiocytes (NRC). We observed unidirectional (i.e., apical to basolateral) Na ϩ -dependent transcellular transport of [ 3 H]taurocholate. Kinetic studies in purified vesicles derived from the apical domain of NRC disclosed saturable Na ϩ -dependent uptake of [ 3 H]taurocholate, with apparent K m and V max values of 209 Ϯ 45 M and 1.23 Ϯ 0.14 nmol/mg/10 s, respectively. Reverse transcriptase PCR (RT-PCR) using degenerate primers for both the rat liver Na ϩdependent taurocholate-cotransporting polypeptide and rat ileal apical Na ϩ -dependent bile acid transporter, designated Ntcp and ASBT, respectively, revealed a 206-bp product in NRC whose sequence was identical to the ASBT. Northern blot analysis demonstrated that the size of the ASBT transcript was identical in NRC, freshly isolated cholangiocytes, and terminal ileum. In situ RT-PCR on normal rat liver showed that the message for ASBT was present only in cholangiocytes. Immunoblots using a well-characterized antibody for the ASBT demonstrated a 48-kD protein present only in apical membranes. Indirect immunohistochemistry revealed apical localization of ASBT in cholangiocytes in normal rat liver. The data provide direct evidence that conjugated bile acids are taken up at the apical domain of cholangiocytes via the ASBT, and are consistent with the notion that cholangiocyte physiology may be directly influenced by bile acids. ( J. Clin. Invest. 1997. 100:2714-2721.) Key words: biliary epithelia • taurocholate • transport • liver • plasma membrane vesicles Preliminary portions of this work were presented at the 47th meeting of the American Association for the Study of Liver Diseases, and have been published in abstract form (1996. Hepatology. 94:897 a ).
Development of active and passive transport of bile acids in rabbit intestine
Mechanisms of Ageing and Development, 1987
Previous studies have indicated that saturable, N absent in the ileum throughout most of the suckling t ileal bile acid uptake which occurs during weaning resl in functional bile acid carriers within the ileal brush 1 undertaken in weanling and adult rabbits to establish active ileal and passive jejunal and colonic uptake of 8 a range of concentrations of cholic (C), taurocholic (T(cholic (CDC), tauroehenodeoxycholic (TCDC), glychoc cholic (I)C) and taurodeoxycholic (TI)C) acid was detl TC, GC, DC and TI)C was greater in adult than in we~)rhea and mat me maturatac results primarily through an inc border membrane. This stud~ establish the effect of maturation ol bile acids. The in vitro upta (TC), glycocholic (GC), chenod¢ rchochenodeoxycholic (GCI)C), de determined. Active ileal uptake ruling animals, whereas uptaJ was similar in both groups. The relative permeability for p~ o the jejunum and colon was similar in young and adult rat area was similar in the two groups, but was greater in the i ,' weanling rabbits due to an increase in villus height, width serosal length. However, the age-associated differences in a not explained simply on the basis of these differences in • the concentration of bile acids in the intestinal lumen, q }it from weanling to adulthood does not influence the rel the jejunum or colon to bile acids, but does increase active td unconjugated cholic acid and deoxycholic acid, but not c[ 1; Aging; Bile acids; Colon; Ileum; Jejunum; Ontogeny; P~
Characterization of the Role of ABCG2 as a Bile Acid Transporter in Liver and Placenta
Molecular Pharmacology, 2011
ABCG2 is involved in epithelial transport/barrier functions. Here, we have investigated its ability to transport bile acids in liver and placenta. Cholylglycylamido fluorescein (CGamF) was exported by WIF-B9/R cells, which do not express the bile salt export pump (BSEP). Sensitivity to typical inhibitors suggested that CGamF export was mainly mediated by ABCG2. In Chinese hamster ovary (CHO cells), coexpression of rat Oatp1a1 and human ABCG2 enhanced the uptake and efflux, respectively, of CGamF, cholic acid (CA), glycoCA (GCA), tauroCA, and taurolithocholic acid-3-sulfate. The ability of ABCG2 to export these bile acids was confirmed by microinjecting them together with inulin in Xenopus laevis oocytes expressing this pump. ABCG2-mediated bile acid transport was inhibited by estradiol 17-D-glucuronide and fumitremorgin C. Placental barrier for bile acids accounted for Ͻ2-fold increase in fetal cholanemia despite Ͼ14-fold increased maternal cholanemia induced by obstructive cholestasis in pregnant rats. In rat placenta, the expression of Abcg2, which was much higher than that of Bsep, was not affected by short-term cholestasis. In pregnant rats, fumitremorgin C did not affect uptake/secretion of GCA by the liver but inhibited its fetal-maternal transfer. Compared with wild-type mice, obstructive cholestasis in pregnant Abcg2(Ϫ/Ϫ) knockout mice induced similar bile acid accumulation in maternal serum but higher accumulation in placenta, fetal serum, and liver. In conclusion, ABCG2 is able to transport bile acids. The importance of this function depends on the relative expression in the same epithelium of other bile acid exporters. Thus, ABCG2 may play a key role in bile acid transport in placenta, as BSEP does in liver.
Cholesterol modulates human intestinal sodium-dependent bile acid transporter
AJP: Gastrointestinal and Liver Physiology, 2005
Bile acids are efficiently absorbed from the intestinal lumen via the ileal apical sodium-dependent bile acid transporter (ASBT). ASBT function is essential for maintenance of cholesterol homeostasis in the body. The molecular mechanisms of the direct effect of cholesterol on human ASBT function and expression are not entirely understood. The present studies were undertaken to establish a suitable in vitro experimental model to study human ASBT function and its regulation by cholesterol. Luminal membrane bile acid transport was evaluated by the measurement of sodium-dependent 3H-labeled taurocholic acid (3H-TC) uptake in human intestinal Caco-2 cell monolayers. The relative abundance of human ASBT (hASBT) mRNA was determined by real-time PCR. Transient transfection and luciferase assay techniques were employed to assess hASBT promoter activity. Caco-2 cell line was found to represent a suitable model to study hASBT function and regulation. 25-Hydroxycholesterol (25-HCH; 2.5 μg/ml fo...
Effect of maternal cholestasis on bile acid transfer across the rat placenta–maternal liver tandem
Hepatology, 2000
Cholestasis of pregnancy induces alterations in bile acid transport by human trophoblast plasma membrane (TPM) vesicles. We investigated whether maternal cholestasis affects the overall ability of the rat placenta to carry out vectorial bile acid transfer from the fetus to the mother. Complete obstructive cholestasis (OCP) was maintained during the last week of pregnancy and released at term (day 21), before experiments were performed. In situ single-pass perfusion of one placenta per rat with 250 nmol [14C]glycocholic acid (GC) revealed an impaired uptake in OCP rats (2.28 vs. 5.53 nmol in control rats). Approximately 100% of GC taken up by control placentas was secreted in maternal bile over 120 minutes (5.38 nmol), whereas this was only 61% (1.40 nmol) of the GC taken up by OCP placentas. When 5 nmol GC was administered through the jugular vein no significant difference between both groups in total GC bile output was found. The efficiency (Vmax /KM ) of adenosine triphosphate (ATP)-dependent GC transport by vesicles from the maternal side of TPM was decreased (−41%) in OCP. Moreover, histological examination of the placentas suggested a reduction in the amount of functional trophoblast in the OCP group. This was consistent with a lower antipyrine diffusion across the placenta in these animals. In sum, our results indicate that maternal cholestasis affects the ability of the placenta to efficiently carry out bile acid transfer from fetal to maternal blood. Changes in both the structure and the functionality of the chorionic tissue may account for this impairment.