Cytoskeletal rearrangement and Src and PI-3K-dependent Akt activation control GABA(B)R-mediated chemotaxis.
Cytoskeletal rearrangement and Src and PI-3K-dependent Akt activation control GABA(B)R-mediated chemotaxis.
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DOI：
10.1016/j.cellsig.2015.02.022
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发表时间：
2015-06
影响因子：
4.8
通讯作者：
Klein JB
中科院分区：
文献类型：
--
作者：
Barati MT;Scherzer J;Wu R;Rane MJ;Klein JB
γ-氨基丁酸（GABA）B型受体（GABABR）作为化学引诱物受体在人中性粒细胞中响应于GABABR激动剂而起作用。本研究的目的是确定信号机制调节GABABR介导的趋化性和细胞骨架重排。在蛋白质组学研究中，我们确定了丝氨酸/苏氨酸激酶Akt，酪氨酸激酶Src和Pyk 2，微管调节因子驱动蛋白和微管亲和力调节激酶（MARK）与GABABR共免疫沉淀。为了确定GABABR介导的趋化性中这些候选信号事件的贡献，我们使用了稳定转染人GABAB 1b和GABAB 2受体的大鼠嗜碱性白血病细胞（RBL-2 H3细胞）。GABABR激动剂巴氯芬通过与其特异性GABABR结合诱导Akt磷酸化和趋化性，因为用GABABR拮抗剂CGP 52432预处理细胞阻断了这种作用。此外，巴氯芬诱导的Akt磷酸化被证明依赖于PI-3 K和Src激酶。巴氯芬未能刺激肌动蛋白聚合悬浮RBL细胞，除非暴露于巴氯芬梯度。然而，巴氯芬刺激肌动蛋白和微管蛋白聚合在贴壁RBL-GABABR细胞。分别用细胞松弛素D或诺考达唑处理细胞，阻断肌动蛋白和微管蛋白聚合，废除了杆菌介导的趋化性。此外，巴氯芬刺激Pyk 2和STAT 3磷酸化，这两种已知的细胞迁移调节剂。总之，GABABR刺激促进RBL细胞中的趋化性，这依赖于通过PI 3-K/Akt、Src激酶的信号传导以及微管和肌动蛋白细胞骨架的重排。这些数据定义了GABABR介导的趋化性的机制，其可潜在地用于治疗性调节对损伤和疾病的细胞反应。
The γ-amino butyric acid (GABA) type B receptors (GABABR) function as chemoattractant receptors in response to GABABR agonists in human neutrophils. The goal of this study was to define signaling mechanisms regulating GABABR-mediated chemotaxis and cytoskeletal rearrangement. In a proteomic study we identified serine/threonine kinase Akt, tyrosine kinases Src and Pyk2, microtubule regulator kinesin and microtubule affinity-regulating kinase (MARK) co-immunoprecipitating with GABABR. To define the contributions of these candidate signaling events in GABABR-mediated chemotaxis, we used rat basophilic leukemic cells (RBL-2H3 cells) stably transfected with human GABAB1b and GABAB2 receptors. The GABABR agonist baclofen induced Akt phosphorylation and chemotaxis by binding to its specific GABABR since pretreatment of cells with CGP52432, a GABABR antagonist, blocked such effects. Moreover, baclofen induced Akt phosphorylation was shown to be dependent upon PI-3K and Src kinases. Baclofen failed to stimulate actin polymerization in suspended RBL cells unless exposed to a baclofen gradient. However, baclofen stimulated both actin and tubulin polymerization in adherent RBL-GABABR cells. Blockade of actin and tubulin polymerization by treatment of cells with cytochalasin D or nocodazole respectively, abolished baclofen-mediated chemotaxis. Furthermore, baclofen stimulated Pyk2 and STAT3 phosphorylation, both known regulators of cell migration. In conclusion, GABABR stimulation promotes chemotaxis in RBL cells which is dependent on signaling via PI3-K/Akt, Src kinases and on rearrangement of both microtubules and actin cytoskeleton. These data define mechanisms of GABABR-mediated chemotaxis which may potentially be used to therapeutically regulate cellular response to injury and disease.
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影响因子：
5.3
作者：
Bruzzaniti, Angela;Neff, Lynn;Baron, Roland
通讯作者：
Baron, Roland
影响因子：
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影响因子：
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