Na,K-ATPase isoform expression in sheep red blood cell precursors

Dhir, Rajan; Nishioka, Yasuhiko; Blostein, Rhoda

doi:10.1016/0005-2736(90)90056-t

Cited by 15 publications

(4 citation statements)

References 11 publications

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“…HK dog red cells were also shown by Western blotting to possess the ␤1 subunit, and reticulocytes from both HK and LK dogs express the ␣1 isoform that decreases on maturation in HK cells but is not discernable in LK cells (7). Messenger RNA obtained from the bone marrows of sheep displaying HK and LK red cell phenotypes were found to express the ␣1 and ␤1 isoforms, but not ␣2, ␣3, or ␤2 isoforms (11). Human bone marrow mRNA has been reported (1) to contain all of the Na pump isoforms except ␣4.…”

mentioning

confidence: 98%

Na pump isoforms in human erythroid progenitor cells and mature erythrocytes

Hoffman

Wickrema

Potapova

et al. 2002

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

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This study is aimed at identifying the Na pump isoform composition of human erythroid precursor cells and mature human erythrocytes. We used purified and synchronously growing human erythroid progenitor cells cultured for 7-14 days. RNA was extracted from the progenitor cells on different days and analyzed by RT-PCR. The results showed that only the ␣1, ␣3, ␤2, and ␤3 subunit isoforms and the ␥ modulator were present. Northern analysis of the erythroid progenitor cells again showed that ␤2 but not ␤1 or ␣2 isoforms were present. The erythroid cells display a unique ␤ subunit expression profile (called ␤-profiling) in that they contain the message for the ␤2 isoform but not ␤1, whereas leukocytes and platelets are known to have the message for the ␤1 but not for the ␤2 isoform. This finding is taken to indicate that our preparations are essentially purely erythroid and free from white cell contamination. Western analysis of these cultured progenitor cells confirmed the presence of ␣1, ␣3, (no ␣2), ␤2, ␤3, and ␥ together now with clear evidence that ␤1 protein was also present at all stages. Western analysis of the Na pump from mature human erythrocyte ghosts, purified by ouabain column chromatography, has also shown that ␣1, ␣3, ␤1, ␤2, ␤3, and ␥ are present. Thus, the Na pump isoform composition of human erythroid precursor cells and mature erythrocytes contains the ␣1 and ␣3 isoforms of the ␣ subunit, the ␤1, ␤2, and ␤3 isoforms of the ␤ subunit, and the ␥ modulator.

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mentioning

confidence: 98%

Na pump isoforms in human erythroid progenitor cells and mature erythrocytes

Hoffman

Wickrema

Potapova

et al. 2002

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

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“…It is generally assumed that ␣1 and ␤1 are the most common isoforms, and the kidney is considered to be a model tissue for ␣1 and ␤1. Northern blot analysis of sheep bone marrow also showed that only ␣1 and ␤1 isoforms could be detected (20). On the other hand, there are indications for the presence in kidney of small amounts of ␣2 and ␣3 (8), ␤2 and ␣3 (21,22), and ␤3 (6).…”

Section: Discussionmentioning

confidence: 96%

“…Because it had been concluded (20,23) on the basis of Northern blotting that the concentration of Na,K-ATPase mRNA in reticulocytes must be very low, we decided to use the PCR method. This method makes it possible to deal with the problem of the extremely low abundance of Na,K-ATPase, and to provide a way to obtain complete sequence information.…”

Section: Discussionmentioning

confidence: 99%

Na,K-ATPase subunit isoforms in human reticulocytes: Evidence from reverse transcription–PCR for the presence of α1, α3, β2, β3, and γ

Stengelin

Hoffman²

1997

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

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The objective of this study has been to determine which Na,K-ATPase isoforms are expressed in red blood cells and whether kinetic differences in the uncoupled sodium eff lux mode between the human red blood cell Na,KATPase and other preparations can be explained by differences in the underlying subunit composition. To this end, human reticulocyte RNA was isolated, reverse transcribed, amplified by PCR and appropriate primers, and sequenced. Primers from highly conserved areas as well as isoformspecific primers were used. The ␣1 and ␣3 isoforms of the ␣ subunit, and the ␤2 and ␤3 isoforms of the ␤ subunit were found. The complete coding regions of the cDNAs for the reticulocyte subunits were sequenced from overlapping PCR fragments. No difference was found between the reticulocyte isoforms and the ones already known. The fact that we found ␤2 but not ␤1 in reticulocyte single-stranded cDNA, and ␤1 but not ␤2 in a leukocyte library indicates that leukocyte contamination of our reticulocyte preparation was negligible. Analysis of a human bone marrow library showed that ␣1, ␣2, and ␣3 as well as all three ␤ isoforms were present. The extent to which the kinetic properties of uncoupled sodium eff lux might depend on different isoform combinations is not yet known.

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“…• Switches in Na + /K + -ATPase α-subunit isoform expressed on the cell membrane, or an isoform specific shift of Na + /K + -ATPase molecules from intracellular pools to the plasma membrane [20,[29][30][31]. Erythrocytes however express essentially only the α1-isoform [32].…”

Section: Introductionmentioning

confidence: 99%

Na/K-ATPase Activity and Ketone Body Metabolism in Long-term Diabetic Rats

Buxbaum

2019

Preprint

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The long-term (34 weeks) effect of streptozotocin induced diabetes was assessed in Wistar rats.Na + /K + -ATPase activity was measured by ouabain inhibitable 86 Rb + -uptake into erythrocytes. No difference in the rate of Rb + -uptake, the K m for Rb + or the K i for ouabain was detected between normal and diabetic rats. Thus, the change in Na + /K + -ATPase activity repeatedly described in short-term studies may not translate into a long term physiologically relevant change in ion flux through the sodium pump.Rats excrete ketone bodies mainly as β-hydroxybutyrate. This compound does not show up with nitroprusside sodium based test sticks, it can however be detected by coupled spectrophotometric assay with hydroxybutyrate dehydrogenase.Almost half of the diabetic animals reverted to a non-diabetic state during the experiment, followed by at least partial reversal of secondary diabetic damage.

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Na,K-ATPase isoform expression in sheep red blood cell precursors

Cited by 15 publications

References 11 publications

Na pump isoforms in human erythroid progenitor cells and mature erythrocytes

Na pump isoforms in human erythroid progenitor cells and mature erythrocytes

Na,K-ATPase subunit isoforms in human reticulocytes: Evidence from reverse transcription–PCR for the presence of α1, α3, β2, β3, and γ

Na/K-ATPase Activity and Ketone Body Metabolism in Long-term Diabetic Rats

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