Constitutive expression of a megakaryocytic functional property by murine erythroleukemia (Friend) cells: the incorporation of serotonin.
Murine Friend-derived erythroleukemia cells (MEL) are generally believed to be unipotential progenitors inducible to terminal erythroid differentiation. However, we found that MEL can constitutively incorporate significant amounts of radiolabeled serotonin ([3H]5-HT). Because this process is typical of cells belonging to the megakaryocytic lineage, we investigated the significance and mechanisms of 5-HT incorporation in the MEL system. We observed that: 1) normal murine erythroid cells and erythroid progenitors do not incorporate [3H]5-HT, as well as normal murine myeloid cells and the human myeloid cell line HL-60; on the other hand, the human erythroleukemia cell lines K562 and HEL, which have been shown to constitutively express megakaryocytic features, were able to incorporate [3H]5-HT; 2) MEL incorporated 5-HT by an active and saturable mechanism, dependent on temperature and sodium concentration in the medium; and 3) 5-HT uptake was very rapid. Moreover, because about 65% of cell-associated radioactivity was no longer displaced by the cold substrate, we assumed it to represent "true" cytoplasmic internalization. Finally, 5-HT incorporation by MEL was inhibited by clomipramine, ouabain, and reserpine, which are known inhibitors of 5-HT uptake in platelets. The commitment of MEL to terminal erythroid differentiation by hexamethylene bisacetamide or dimethyl sulfoxide greatly reduced the capacity to incorporate [3H]5-HT. These results seem to suggest that the MEL system, although mainly erythroid as regards its differentiation capability, constitutively expresses features of the megakaryocytic lineage, possibly disclosed by the ability to incorporate 5-HT. This hypothesis was further supported by the findings that 30%-40% of uninduced MEL were labeled by a polyclonal antibody raised against murine platelets that selectively recognized megakaryocytes in murine bone marrow smears.
Vannucchi
A M
AM
Division of Hematology, University of Florence, USL, Italy.
Grossi
A
A
Paoletti
F
F
Rafanelli
D
D
Bacci
P
P
Ferrini
P R
PR
D000081:Acetamides / Q000494:pharmacology; D000818:Animals; D001693:Biological Transport, Active; D002997:Clomipramine / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D015672:Erythroid Precursor Cells / Q000378:metabolism; D005622:Friend murine leukemia virus; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism*; D008533:Megakaryocytes / Q000378:metabolism*; D051379:Mice; D010042:Ouabain / Q000494:pharmacology; D012110:Reserpine / Q000494:pharmacology; D012701:Serotonin / Q000378:metabolism*; D012964:Sodium / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000081:Acetamides; D012701:Serotonin; D010042:Ouabain; D012110:Reserpine; D012964:Sodium; C014026:hexamethylene bisacetamide; D002997:Clomipramine