Amino acid deprivation-induced stress response in the bovine renal epithelial cell line NBL-1: Induction of HSP 70 by phenylalanine

The glutamate transport system of the bovine renal epithelial cell line NBL-1 was studied. The Km for Na(+)-dependent glutamate transport was found to be 13.8 +/- 2.4 microM (Vmax. 365 +/- 19.2 pmol/3 min per mg) and for Na(+)-dependent aspartate transport 4.5 +/- 1.1 microM (Vmax. 108 +/- 6.3 pmol/3 min per mg). The Km values are in close agreement with those expected for high-affinity Na(+)-dependent glutamate transport by System XAG-. Upon deprivation of amino acids, the Vmax. for Na+/aspartate co-transport rose to 203 +/- 6.0 pmol/3 min per mg (Km 3.8 +/- 0.5 microns). A probe was constructed to the high-affinity excitatory amino acid carrier (EAAC1) [Kanai and Hediger (1992) Nature (London) 360, 467-471]. The probe hybridized to a 3.5 kb transcript. On deprivation of amino acids, the level of EAAC1 mRNA decreased sharply before the measurable increase in transport levels, but was subsequently restored to control levels. A motif, which we propose is linked to amino acid deprivation, was found in the EAAC1 primary sequence.

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Regulation of Na+-Dependent Amino Acid Transport in the Bovine Renal Epithelial Cell Line NBL-1

Contributions to Nephrology - Renal Ammoniagenesis Interorgan Cooperation in Acid- Base Homeostasis ◽

10.1159/000423405 ◽

2015 ◽

pp. 103-108

Author(s):

J. D. McGivan ◽

S. Plakidou-Dymock

Keyword(s):

Amino Acid ◽

Epithelial Cell ◽

Cell Line ◽

Amino Acid Transport ◽

Epithelial Cell Line ◽

Acid Transport ◽

Renal Epithelial Cell ◽

Renal Epithelial Cell Line

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Amino acid deprivation leads to the emergence of System A activity and the synthesis of a specific membrane glycoprotein in the bovine renal epithelial cell line NBL-1

Biochemical Journal ◽

10.1042/bj2840577 ◽

1992 ◽

Vol 284 (2) ◽

pp. 577-582 ◽

Cited By ~ 21

Author(s):

A Felipe ◽

C Soler ◽

J D McGivan

Keyword(s):

Amino Acid ◽

Epithelial Cell ◽

Epithelial Cell Line ◽

Membrane Glycoprotein ◽

Amino Acid Deprivation ◽

Transport Activity ◽

Renal Epithelial Cell ◽

System A ◽

Alanine Transport ◽

Renal Epithelial Cell Line

1. Amino acid deprivation of confluent monolayers of the bovine renal epithelial cell line NBL-1 causes a stimulation of Na(+)-dependent alanine transport. 2. This stimulation is mediated by a protein-synthesis-dependent induction of 2-(methylamino)isobutyric acid (methyl-AIB)-sensitive alanine transport activity (System A), which was not previously present in these cells. 3. Induction was prevented by the addition of methyl-AIB, alanine or glutamine. 4. Tunicamycin prevented the induction of alanine transport activity. 5. Induction of System A activity was accompanied by incorporation of [3H]mannose into a single membrane protein band of molecular mass 113-140 kDa. 6. These results are consistent with the possibility that induced System A activity in confluent NBL-1 cells is mediated by the synthesis of a 113-140 kDa membrane glycoprotein.

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Electrophysiological Measurements of a Toad Renal Epithelial Cell Line (A6) as an Assay for Predicting Ocular Eye Irritancy

Alternatives to Laboratory Animals ◽

10.1177/026119299202000206 ◽

1992 ◽

Vol 20 (2) ◽

pp. 218-221

Author(s):

Henning F. Bjerregaard

Keyword(s):

Epithelial Cell ◽

Cell Line ◽

South African ◽

Mdck Cells ◽

Epithelial Cell Line ◽

Renal Epithelial Cell ◽

Electrophysiological Measurements ◽

Renal Epithelial Cell Line ◽

Darby Canine Kidney

An established epithelial cell line (A6) from a South African clawed toad (Xenopus laevis) kidney was used as a model for the corneal epithelium of the eye in order to determine ocular irritancy. When grown on Millipore filter inserts, A6 cells form a monolayer epithelium of high electrical resistance and generate a trans-epithelial potential difference. These two easily-measured electrophysiological endpoints showed a dose-related decrease after exposure for 24 hours to seven selected chemicals of different ocular irritancy potential. It was demonstrated that both trans-epithelial resistance and potential ranked closely with in vivo eye irritancy data and correlated well (r = 0.96) with loss of trans-epithelial impermeability of Madin-Darby canine kidney (MDCK) cells, detected by use of a fluorescein leakage assay.

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Polarized apical distribution of glycosyl-phosphatidylinositol-anchored proteins in a renal epithelial cell line.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.85.24.9557 ◽

1988 ◽

Vol 85 (24) ◽

pp. 9557-9561 ◽

Cited By ~ 222

Author(s):

M. P. Lisanti ◽

M. Sargiacomo ◽

L. Graeve ◽

A. R. Saltiel ◽

E. Rodriguez-Boulan

Keyword(s):

Epithelial Cell ◽

Cell Line ◽

Epithelial Cell Line ◽

Renal Epithelial Cell ◽

Glycosyl Phosphatidylinositol ◽

Renal Epithelial Cell Line

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Expression of phosphoenolpyruvate carboxykinase (PEPCK) chimeras in renal epithelial cells. Retention of appropriate physiological responsiveness using enhancerless retroviral vectors

Biochemical Journal ◽

10.1042/bj2840725 ◽

1992 ◽

Vol 284 (3) ◽

pp. 725-732 ◽

Cited By ~ 2

Author(s):

A S Pollock ◽

D H Lovett

Keyword(s):

Epithelial Cell ◽

Cyclic Amp ◽

Cell Line ◽

Phosphoenolpyruvate Carboxykinase ◽

Large Fraction ◽

Expression System ◽

Retroviral Vectors ◽

Epithelial Cell Line ◽

Renal Epithelial Cell ◽

Renal Epithelial Cell Line

We used an enhancerless U3 mutant retroviral vector to deliver chimeras of the phosphoenolpyruvate carboxykinase (PEPCK) promoter region to a renal epithelial cell line capable of expressing PEPCK mRNA. Chimeras consisting of the PEPCK promoter and chloramphenicol acetyltransferase, neomycin phosphotransferase or human growth hormone genes were expressed after viral infection of the NRK52E renal epithelial cell line. Virus-delivered sequences in which the direction of PEPCK promoter transcription was antegrade to the normal direction of the long terminal repeat (LTR)-initiated transcription correctly upon stimulation with dexamethasone or 8-bromo cyclic AMP and upon lowering of the extracellular pH. Fluorescent primer extension in situ using primers specific for virus-delivered sequences of antegrade constructs indicated that a large fraction of NRK52E cells could be infected by co-cultivation with virus-producing psi-2 cells without G418 selection. Virus-delivered constructs whose orientation was opposite to that of the LTRs were expressed at very low levels, with transcripts detectable by PCR only in RNA from cyclic AMP-treated cells. Using reverse transcription/PCR, we demonstrated that the chimeric transcripts were from the internal PEPCK promoter rather than a functional or reconstituted Moloney LTR. PEPCK-reporter chimeras delivered by retroviral vectors demonstrated a level of expression more consistent with the level of expression of the native PEPCK gene than did transfected chimeras. This expression system should prove useful for studies of the physiological modulation of gene expression in renal tissues.

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