Identification of a New Member of Type I Na/Phosphate Co-Transporter in the Rat Kidney

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Abstract

Aim: To clone new phosphate transporters from the kidney. Methods: EST bank was screened for new members of Na-dependent inorganic phosphate (Pi) co-transporter. To obtain a full-length clone, a rat kidney cDNA library was screened with the EST clone as a probe. Results: We isolated a cDNA that encodes a type I Na/Pi co-transporter. Based on the sequence homology and tissue distribution, it will be the rat ortholog of human NPT4 (Solute Carrier family 17, member A3, SLC17A3) and is designated rNPT4. When expressed in Xenopus oocytes, Na/Pi co-transport activity was demonstrated. In Northern blotting analysis, rNPT4 was selectively expressed in the liver and kidney. In immunohistochemical analysis, rNPT4 was localized at the apical membrane of the proximal tubules (S2, S3) in the rat kidney. The electron microscopic studies confirmed this finding. Conclusion: The present study revealed the expression of a new type I Na/Pi co-transporter at the brush border membrane of the proximal tubules in the kidney.

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Identification of a vesicular glutamate transporter that defines a glutamatergic phenotype in neurons.

S Takamori, J Rhee, C Rosenmund … (2000)

Glutamate is the major excitatory neurotransmitter in the mammalian central nervous system. Synaptic vesicles are loaded with neurotransmitter by means of specific vesicular transporters. Here we show that expression of BNPI, a vesicle-bound transporter associated with sodium-dependent phosphate transport, results in glutamate uptake by intracellular vesicles. Substrate specificity and energy dependence are very similar to glutamate uptake by synaptic vesicles. Stimulation of exocytosis--fusion of the vesicles with the cell membrane and release of their contents--resulted in quantal release of glutamate from BNPI-expressing cells. Furthermore, we expressed BNPI in neurons containing GABA (gamma-aminobutyric acid) and maintained them as cultures of single, isolated neurons that form synapses to themselves. After stimulation of these neurons, a component of the postsynaptic current is mediated by glutamate as it is blocked by a combination of the glutamate receptor antagonists, but is insensitive to a GABA(A) receptor antagonist. We conclude that BNPI functions as vesicular glutamate transporter and that expression of BNPI suffices to define a glutamatergic phenotype in neurons.

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Targeted inactivation of Npt2 in mice leads to severe renal phosphate wasting, hypercalciuria, and skeletal abnormalities.

Jennifer Beck, H Ozawa, A Karaplis … (1998)

Npt2 encodes a renal-specific, brush-border membrane Na+-phosphate (Pi) cotransporter that is expressed in the proximal tubule where the bulk of filtered Pi is reabsorbed. Mice deficient in the Npt2 gene were generated by targeted mutagenesis to define the role of Npt2 in the overall maintenance of Pi homeostasis, determine its impact on skeletal development, and clarify its relationship to autosomal disorders of renal Pi reabsorption in humans. Homozygous mutants (Npt2(-/-)) exhibit increased urinary Pi excretion, hypophosphatemia, an appropriate elevation in the serum concentration of 1,25-dihydroxyvitamin D with attendant hypercalcemia, hypercalciuria and decreased serum parathyroid hormone levels, and increased serum alkaline phosphatase activity. These biochemical features are typical of patients with hereditary hypophosphatemic rickets with hypercalciuria (HHRH), a Mendelian disorder of renal Pi reabsorption. However, unlike HHRH patients, Npt2(-/-) mice do not have rickets or osteomalacia. At weaning, Npt2(-/-) mice have poorly developed trabecular bone and retarded secondary ossification, but, with increasing age, there is a dramatic reversal and eventual overcompensation of the skeletal phenotype. Our findings demonstrate that Npt2 is a major regulator of Pi homeostasis and necessary for normal skeletal development.

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Uptake of glutamate into synaptic vesicles by an inorganic phosphate transporter.

E Bellocchio, R J Reimer, R T Fremeau … (2000)

Previous work has identified two families of proteins that transport classical neurotransmitters into synaptic vesicles, but the protein responsible for vesicular transport of the principal excitatory transmitter glutamate has remained unknown. We demonstrate that a protein that is unrelated to any known neurotransmitter transporters and that was previously suggested to mediate the Na(+)-dependent uptake of inorganic phosphate across the plasma membrane transports glutamate into synaptic vesicles. In addition, we show that this vesicular glutamate transporter, VGLUT1, exhibits a conductance for chloride that is blocked by glutamate.

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Author and article information

Journal

Journal ID (publisher-id): NEP

Journal ID (nlm-ta): Nephron Physiol

Journal ID (doi): 10.1159/issn.1660-2137

Title: Nephron Physiology

Publisher: S. Karger AG

ISSN (Electronic): 1660-2137

Publication date Subscription-year: 2003

Publication date (Print): May 2003

Publication date (Electronic): 17 June 2003

Volume: 94

Issue: 1

Pages: p10-p18

Affiliations

^aDepartmentof Pharmacology, Jichi Medical School, Tochigi, and ^bDepartment of Anatomy, School of Medicine, Gunma University, Maebashi, Gunma, Japan

Article

Publisher ID: 71070 Other ID: Nephron Physiol 2003;94:p10–p18

DOI: 10.1159/000071070

PubMed ID: 12806205

SO-VID: cd2e557e-6df7-4078-8162-4a838b0d8b1a

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Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

History

Date received : 07 October 2002

Date accepted : 28 February 2003

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Figures: 7, References: 23, Pages: 1

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Identification of a New Member of Type I Na/Phosphate Co-Transporter in the Rat Kidney

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Abstract

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Karger: Nephrology

Most cited references 8

Identification of a vesicular glutamate transporter that defines a glutamatergic phenotype in neurons.

Targeted inactivation of Npt2 in mice leads to severe renal phosphate wasting, hypercalciuria, and skeletal abnormalities.

Uptake of glutamate into synaptic vesicles by an inorganic phosphate transporter.

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