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DOI: 10.1055/s-0037-1615112
Maturation of Megakaryoblastic Cells Is Accompanied by Upregulation of GSα-L Subtype and Increased cAMP Accumulation
Publication History
Received
27 August 1997
Accepted after resubmission
21 January 1998
Publication Date:
07 December 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199805/lookinside/thumbnails/10-1055-s-0037-1615112-1.jpg)
Summary
In platelets and megakaryoblasts Gs, the trimeric G-protein that stimulates adenylyl cyclase, is present in a short, 45 kDa, and a long, 52 kDa isoform termed Gsα-S and Gsα-L, respectively. To assess the relative contribution of these isoforms in the cellular synthesis of cAMP, the ratio Gsα-S/Gsα-L was changed in the megakaryoblastic cell line DAMI by inducing cell maturation with recombinant human thrombopoietin (TPO) or the phorbol ester PMA. Flow cytometric analysis confirmed that this treatment induced a moderate (TPO) and extensive (PMA) increase in nuclear ploidy and expression of the glycoproteins-IIIa and -Ib. Northern blot analysis revealed down-regulation of total GS-mRNA after treatment of DAMI-cells with TPO and PMA. Western blot analysis showed significant (P <0.05) upregulation of Gs-L with respective amounts of 27 ± 4% of total G s in untreated cells, 35 ± 1% in TPO- and 41 ± 3% in PMA-treated DAMI cells (n = 3-4). DAMI cells contained 6 ± 1 pmol cAMP/106 cells, which was not changed by treatment with TPO or PMA. In untreated cells this level increased to 70 ± 9 pmol cAMP/106 cells after 10 min stimulation with 1 μmol/l of the stable prostacyclin analog iloprost. The same stimulation with iloprost resulted in 165 ± 32 pmol cAMP/106 in TPO-treated cells and in 588 ± 100 pmol cAMP/106 in cells treated with PMA. Thus, a shift from Gsα-S to Gsα-L during megakaryoblast maturation strongly potentiates the production of cAMP. A similar shift may occur during normal megakaryocyte maturation and may explain the extreme sensitivity to prostacyclin of platelets, which contain Gsα-S and Gsα-L in approximately equal amounts.
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