Isolation and characterization of the brush border fraction from newborn rat renal proximal tubule cells

Goldmann, David R.; Schlesinger, Harvey; Segal, Stanton

doi:10.1016/0005-2736(76)90351-5

Cited by 52 publications

(22 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The advent of methods to prepare purified brush border membrane vesicles (12)(13)(14) has permitted studies of transport systems under defined conditions that reflect solute and membrane carrier interactions at one renal tubule reabsorptive surface. Although reports have appeared characterizing the transport of many solutes by renal brush border membranes from adult animals, little is known about the function of renal membranes from newborn and young animals despite the description of their preparation from rat kidney by Qoldmann et al a decade ago (19).…”

Section: Discussionmentioning

confidence: 99%

Developmental aspects of proline transport in rat renal brush border membranes.

Medow¹,

Roth²,

Goldmann³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Proline uptake by rat renal brush border membrane vesicles from animals 7 days of age and older has been examined to delineate developmental changes in membrane function that may underlie the physiological hyperprolinuria of young animals. Although the two proline transport systems normally present in adult membranes were found in membranes from young animals, the proline "overshoot" resulting from a sodium ion gradient is minimal and increases with age of the animal from which the membranes were isolated. This is associated with a severalfold faster entry of 22Na into vesicles of the 7-day-old animal compared to entry into membranes prepared from adult kidneys. The very rapid dissipation of the sodium gradient thus diminishing the driving force for transmembrane proline movement may explain the changes in proline overshoot observed in membranes from young animals. The altered sodium permeability is consistent with the fact that young animals have a generalized inability to reabsorb other amino acids whose transport is known to be sodium gradient stimulated.Urinary hyperexcretion of proline and indeed of most amino acids is characteristic ofneonatal rat (1), dog (2), and man (3). Subsequent postnatal development of the nephron, however, reduces urinary amounts of these acids to much lower levels (4, 5). Although the subject of numerous investigations (6-9), the nature of the developmental changes occurring in the kidney that result in decreased amino acid excretion is as yet undefined. An understanding of these changes should not only increase our knowledge of normal transport processes but also provide insight into human disorders of renal amino acid transport, such as familial iminoglycinuria, in which the affected individual continues to excrete excessive quantities of certain amino acids in the urine.Previous studies of the development of renal amino acid transport in rat have relied upon the use of the cortical slice (10, 11) and isolated renal tubule fragment (1,8). Data obtained from these models, however, are influenced by factors such as bidirectional solute movement, substrate penetration, and substrate metabolism. The use of isolated membrane vesicles obviates these problems and age-related differences in solute transport by these vesicles may be examined directly.In our recent studies, proline uptake by renal brush border membrane vesicles prepared from newborn rats by free-flow electrophoresis, unlike vesicles from adult animals, did not show a Na+-gradient-stimulated overshoot but did exhibit a Na+-gradient-enhanced rate of early proline entry (12). To better understand the events accompanying the maturation of renal solute transport we have therefore examined characteristics of proline and Na+ uptake by renal brush border membrane vesicles prepared from rats of various ages ranging from 1 week oflife to adulthood. The results indicate that, although the two proline transport systems normally present in adult membranes are also found in membranes from young animals, the proline overshoot re...

show abstract

Section: Discussionmentioning

confidence: 99%

Developmental aspects of proline transport in rat renal brush border membranes.

Medow¹,

Roth²,

Goldmann³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…G-6-P activity in these isolates was approximately 60% that reported for rat cortical homogenates [37]and was the only enzyme measured which exhibited no significant change throughout the 28 days of culture. One can only speculate whether this maintenance of activity in culture reflects the multifunctional nature of this enzyme [38]. The fact that FBP and G-6-P were measurable in primary isolates and in monolayer cultures supports the conclusion that these cells originated from the proximal portion of the nephron.…”

Section: Discussionmentioning

confidence: 77%

Isolation, Culture and Characterization of Human Renal Proximal Tubule and Collecting Duct Cells

Trifillis¹

1999

Nephron Exp Nephrol

View full text Add to dashboard Cite

The complexity and heterogeneity of the human nephron with regard to cell types make well-defined in vitro systems of renal cells valuable for studies of the pathogenetic mechanisms involved in nephrotoxicity. In our laboratory renal proximal tubule cells (PTC) and collecting duct cells (CDC) have been isolated, cultured and characterized from cadaver kidneys (postmortem time <24 h) for use in studies of renal cytotoxicity induced by therapeutics and bacteria. PTC seeded at 10⁶ cells/ml formed confluent monolayers within 7 days. Histochemical markers were used to determine the origin of the cell cultures. Cells were negative for factor VIII, positive for cytokeratin and γ-glutamyltransferase (GGT), and bound winged pea lectin. CDC, isolated from the renal papillae, formed monolayers within 14 days of seeding. CDC were negative for factor VIII and GGT, positive for cytokeratin and bound peanut lectin. PTC and CDC isolates and cultures exhibited typical epithelial cell ultrastructure: cell junctions, intermediate filaments, microvilli, and numerous mitochondria. The morphological and histochemical evidence confirms that PTC and CDC can be isolated and cultured for use in in vitro studies.

show abstract

“…2, table II). Identical postnatal development of rat kidney brush-border mem branes was found by Goldmann et al [6],…”

Section: Discussionmentioning

confidence: 80%

Postnatal Development of Rat Kidney Plasma Membrane Na-K-ATPase

Dobrović-Jenik¹,

Ozegović²,

Milković³

1984

Neonatology

View full text Add to dashboard Cite

The postnatal development of the sodium transport mechanism in the rat kidney, as judged by the activity of renal plasma membrane Na-K-ATPase, was studied. Highly purified kidney plasma membranes as verified by electron microscopic and enzyme examinations, were used. Na-K-ATPase activity (μmol Pi/mg protein/h) increases exponentially from 16.9 ± 1.94 found at birth to the mature level of 43.1 ± 2.16 reached at the age of 41 days. This finding paralleled our results of SDS-polyacrylamide gel disc electrophoresis, showing a completely differentiated plasma membrane protein structure at birth, the maturation of which proceeds only as quantitative enrichment of some protein fractions in the further postnatal period.

show abstract

Isolation and characterization of the brush border fraction from newborn rat renal proximal tubule cells

Cited by 52 publications

References 31 publications

Developmental aspects of proline transport in rat renal brush border membranes.

Developmental aspects of proline transport in rat renal brush border membranes.

Isolation, Culture and Characterization of Human Renal Proximal Tubule and Collecting Duct Cells

Postnatal Development of Rat Kidney Plasma Membrane Na-K-ATPase

Contact Info

Product

Resources

About