Distinct functional effects for dynamin 3 during megakaryocytopoiesis.
Distinct functional effects for dynamin 3 during megakaryocytopoiesis.
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动力蛋白 3 在巨核细胞生成过程中具有独特的功能作用。
DOI：
10.1089/scd.2011.0159
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发表时间：
2011
影响因子：
4
通讯作者：
Reems,Jo-Anna
中科院分区：
文献类型：
--
作者：
Wang,Wenjing;Gilligan,DianaM;Sun,Sijie;Wu,Xiaoping;Reems,Jo-Anna
动力蛋白3（DNM3）是参与许多膜重排如胞质分裂、运输囊泡的出芽、吞噬作用和细胞运动的运动蛋白家族的成员。最近，DNM3被认为在巨核细胞（MK）发育中起作用。为了进一步研究DNM3在巨核细胞生成过程中的功能作用，我们将序列特异性短发夹RNA（shRNA）引入发育中的MK中。结果表明，DNM3的敲低抑制了涉及祖细胞扩增的MK发育阶段。这通过巨核细胞集落形成单位数量、有核细胞总数以及培养物中产生的CD41+和CD61+ MK数量的显著减少而得到证实。使用苯乙烯膜染料定量终末分化的MK的分界膜系统（DMS），我们发现DNM3与DMS共定位，并且DNM3慢病毒shRNA阻止DMS的形成。敲低鼠MK中的发动蛋白3也导致形态学上大的MK的数量减少，并且相对于对照，大的MK的总体尺寸减小。MK蛋白裂解物用于重叠印迹，以显示DNM3和肌动蛋白均与非肌肉肌球蛋白IIA（MYH9）结合。与这些观察结果一致，MK和前血小板过程的免疫荧光研究显示DNM3与MYH9共定位。总之，这些研究表明，DNM3不仅参与MK祖细胞扩增，而且还参与细胞质扩大和DMS的形成。
Dynamin 3 (DNM3) is a member of a family of motor proteins that participate in a number of membrane rearrangements such as cytokinesis, budding of transport vesicles, phagocytosis, and cell motility. Recently, DNM3 was implicated as having a role in megakaryocyte (MK) development. To further investigate the functional role of DNM3 during megakaryocytopoiesis, we introduced sequence-specific short hairpin RNAs (shRNAs) into developing MKs. The results showed that knockdown of DNM3 inhibited a stage of MK development that involved progenitor amplification. This was evident by significant decreases in the number of colony forming unit-megakaryocytes, the total number of nucleated cells, and the number of CD41+and CD61+MKs produced in culture. Using a styrl membrane dye to quantify the demarcation membrane system (DMS) of terminally differentiated MKs, we found that DNM3 co-localized with the DMS and thatDNM3lentiviral shRNAs precluded the formation of the DMS. Knockdown of dynamin 3 in murine MKs also caused a decrease in the number of morphologically large MKs and the overall size of large MKs was decreased relative to controls. MK protein lysates were used in overlay blots to show that both DNM3 and actin bind to nonmuscle myosin IIA (MYH9). Consistent with these observations, immunofluorescence studies of MKs and proplatelet processes showed co-localization of DNM3 with MYH9. Overall, these studies demonstrate that DNM3 not only participates in MK progenitor amplification, but is also involved in cytoplasmic enlargement and the formation of the DMS.