Schick P K, Weinstein M
J Lab Clin Med. 1981 Oct;98(4):607-15.
Serotonin accumulation was studied in guinea pig megakaryocytes and platelets. Megakaryocyte suspensions, 80% to 90% pure, and platelet-rich plasma were used. 14C-serotonin was used to assay serotonin uptake. Maximum capacity for serotonin storage in 2.5 x 10(5) megakaryocytes was 4.2 nmol and in 2.5 x 10(8) platelets was 3.1 nmol. One thousand times as many platelets as megakaryocytes were used to normalize for protein content of the two cells. Thus the megakaryocyte's capacity for accumulating serotonin is equivalent to, if not greater than, that of the platelet. Serotonin accumulation in individual megakaryocytes and other hematopoietic cells was studied by autoradiography. Whole marrow suspensions were incubated with 3H-serotonin. Cells in whole marrow suspensions, cells in the pellet of the albumin density gradient, and purified megakaryocytes were then analyzed by autoradiography. The pellet of the albumin density gradient was inspected because it is known to contain megakaryocytes that are not recovered in the purified megakaryocyte suspensions. Examination of whole marrow cell suspensions revealed that virtually all megakaryocytes had taken up serotonin, but there was no evidence of serotonin accumulation in other hematopoietic cells. Over 95% of megakaryocytes in the pellet of the albumin density gradient had retained serotonin. The megakaryocytes that had not retained serotonin or not accumulated the amine did not comprise a specific population in respect to maturity. Evaluation of purified megakaryocytes showed that megakaryocytes at various stages of maturation, as defined by diverse size and cytoplasmic/nuclear maturation, had accumulated equivalent amounts of serotonin. Thus the capacity for serotonin accumulation is established in the immature megakaryocyte. Serotonin uptake can serve as a marker for the identification of the immature megakaryocyte.
在豚鼠巨核细胞和血小板中研究了5-羟色胺的蓄积情况。使用了纯度为80%至90%的巨核细胞悬液和富含血小板的血浆。用14C-5-羟色胺来测定5-羟色胺摄取量。2.5×10(5)个巨核细胞中5-羟色胺的最大储存量为4.2 nmol,2.5×10(8)个血小板中为3.1 nmol。使用的血小板数量是巨核细胞的一千倍,以校正这两种细胞的蛋白质含量。因此,巨核细胞蓄积5-羟色胺的能力即使不大于血小板,也与之相当。通过放射自显影研究了单个巨核细胞和其他造血细胞中5-羟色胺的蓄积情况。将全骨髓悬液与3H-5-羟色胺一起孵育。然后通过放射自显影分析全骨髓悬液中的细胞、白蛋白密度梯度沉淀中的细胞以及纯化的巨核细胞。检查白蛋白密度梯度沉淀是因为已知其中含有在纯化的巨核细胞悬液中未回收的巨核细胞。对全骨髓细胞悬液的检查显示,几乎所有巨核细胞都摄取了5-羟色胺,但没有证据表明其他造血细胞中有5-羟色胺蓄积。白蛋白密度梯度沉淀中的巨核细胞超过95%保留了5-羟色胺。未保留5-羟色胺或未蓄积该胺的巨核细胞在成熟度方面并不构成一个特定群体。对纯化的巨核细胞的评估表明,根据不同大小和细胞质/细胞核成熟度定义的不同成熟阶段的巨核细胞蓄积了等量的5-羟色胺。因此,未成熟巨核细胞中确立了蓄积5-羟色胺的能力。5-羟色胺摄取可作为鉴定未成熟巨核细胞的标志物。