Activation of the neurokinin 3 receptor promotes filopodia growth and sprouting in rat embryonic hypothalamic cells.
Activation of the neurokinin 3 receptor promotes filopodia growth and sprouting in rat embryonic hypothalamic cells.
复制标题
神经激肽 3 受体的激活促进大鼠胚胎下丘脑细胞中丝状伪足的生长和发芽。
DOI：
10.1002/dneu.22207
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发表时间：
2015
影响因子：
3
通讯作者：
Thakar,Amit
中科院分区：
文献类型：
--
作者：
Flynn,FrancisW;Kinney-Lang,Eli;Hoekstra,Chelsea;Pratt,DonaldL;Thakar,Amit
速激肽家族的成员对发育中的神经元具有营养作用。速激肽神经激肽3受体（NK 3R）出现在胚胎发育的早期;在下丘脑神经元的高峰出生期，但其参与神经发育尚未检查。为了解决其可能的作用，永生化的胚胎下丘脑神经元（CLU 209）用CellMask（一种质膜染色剂）处理，或者在用pEGFP-NK 3R表达载体转染的CLU 209细胞中对膜进行成像。然后用senktide（一种NK 3 R激动剂）或Dulbecco改良Eagle培养基（DMEM）处理未转染细胞和转染细胞，并在随后的30 min内捕获延时共聚焦图像。与DMEM相比，senktide处理导致未转染和转染的CLU 209细胞的索马体开始出现丝状伪足。这些丝状伪足的直径和长度分别约为200 nm和3 µm。用IP 3受体阻断剂2-氨基乙氧基二苯基硼酸盐（2-APB）预处理，阻止了senktide诱导的丝状伪足生长;表明NK 3R诱导的丝状伪足生长可能涉及细胞内钙的释放。将转染的CLU 209细胞暴露于senktide 24 h导致丝状伪足和延伸10-20 µm的突起进一步生长。建立了一个由线性模型和种群模型组成的数学模型，用于描述在几分钟的时间尺度内丝状伪足生长的动态。结果表明，配体诱导的NK 3R激活通过启动丝状伪足形成来影响早期发育过程，丝状伪足形成是轴突发生的先决条件。© 2014 Wiley Periodicals，Inc.开发神经生物学75：12-22，2015
Members of the tachykinin family have trophic effects on developing neurons. The tachykinin neurokinin 3 receptor (NK3R) appears early in embryonic development; during the peak birthdates of hypothalamic neurons, but its involvement in neural development has not been examined. To address its possible role, immortalized embryonic hypothalamic neurons (CLU209) were treated with CellMask, a plasma membrane stain, or the membranes were imaged in CLU209 cells that were transfected with a pEGFP‐NK3R expression vector. Nontransfected cells and transfected cells were then treated with senktide, a NK3R agonist, or Dulbecco's Modified Eagle's Medium (DMEM) and time‐lapse confocal images were captured for the following 30 min. Compared to DMEM, senktide treatment led to filopodia initiation from the soma of both nontransfected and transfected CLU209 cells. These filopodia had diameters and lengths of approximately 200 nm and 3 µm, respectively. Pretreatment with an IP3 receptor blocker, 2‐aminoethoxydiphenyl borate (2‐APB), prevented the senktide‐induced growth in filopodia; demonstrating that NK3R‐induced outgrowth of filopodia likely involves the release of intracellular calcium. Exposure of transfected CLU209 cells to senktide for 24 h led to further growth of filopodia and processes that extended 10–20 µm. A mathematical model, composed of a linear and population model was developed to account for the dynamics of filopodia growth during a timescale of minutes. The results suggest that the ligand‐induced activation of NK3R affects early developmental processes by initiating filopodia formation that are a prerequisite for neuritogenesis. © 2014 Wiley Periodicals, Inc. Develop Neurobiol 75: 12–22, 2015