Developmental stage-specific distribution and phosphorylation of Mblk-1, a transcription factor involved in ecdysteroid-signaling in the honey bee brain
Developmental stage-specific distribution and phosphorylation of Mblk-1, a transcription factor involved in ecdysteroid-signaling in the honey bee brain
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DOI：
10.1038/s41598-020-65327-z
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发表时间：
2020-05-26
影响因子：
4.6
通讯作者：
Kubo, Takeo
中科院分区：
文献类型：
--
作者：
Kumagai, Hitomi;Kunieda, Takekazu;Kubo, Takeo
在蜜蜂中，蘑菇体（MB）是昆虫脑中的高级中心，包括称为Kenyon细胞（KC）的中间神经元。我们先前报道了Mblk-1，其编码参与蜕皮激素信号传导的转录因子，优先在蛹和成虫工蜂脑中的大型KC（lKC）中表达，并且Ras/MAPK途径的磷酸化增强了Mblk-1的体外转录活性。在本研究中，我们使用亲和纯化的抗Mblk-1和抗磷酸化Mblk-1抗体进行免疫印迹和免疫荧光研究，以分析Mblk-1在蛹和成年工蚁大脑中的分布和磷酸化。Mblk-1在蛹和成虫工蜂脑中优先表达于IKC中。与此相反，一些Mblk-1磷酸化几乎完全在蛹阶段，和磷酸化的Mblk-1优先表达在MB成神经细胞和IKCs蛹的大脑。免疫荧光研究表明，Mblk-1和磷酸化的Mblk-1都位于蛹和成虫工蜂脑中lKC胞体的细胞质和细胞核中。这些发现表明，Mblk-1在蛹和成虫阶段的IKC中起作用，并且磷酸化的Mblk-1在蜜蜂脑中的MB成神经细胞和IKC中具有蛹阶段特异性功能。
In the honey bee, the mushroom bodies (MBs), a higher-order center in insect brain, comprise interneurons termed Kenyon cells (KCs). We previously reported that Mblk-1, which encodes a transcription factor involved in ecdysteroid-signaling, is expressed preferentially in the largetype KCs (lKCs) in the pupal and adult worker brain and that phosphorylation by the Ras/MAPK pathway enhances the transcriptional activity of Mblk-1 in vitro. In the present study, we performed immunoblotting and immunofluorescence studies using affinity-purified anti-Mblk-1 and anti-phosphorylated Mblk-1 antibodies to analyze the distribution and phosphorylation of Mblk-1 in the brains of pupal and adult workers. Mblk-1 was preferentially expressed in the lKCs in both pupal and adult worker brains. In contrast, some Mblk-1 was phosphorylated almost exclusively in the pupal stages, and phosphorylated Mblk-1 was preferentially expressed in the MB neuroblasts and lKCs in pupal brains. Immunofluorescence studies revealed that both Mblk-1 and phosphorylated Mblk-1 are located in both the cytoplasm and nuclei of the lKC somata in the pupal and adult worker brains. These findings suggest that Mblk-1 plays a role in the lKCs in both pupal and adult stages and that phosphorylated Mblk-1 has pupal stage-specific functions in the MB neuroblasts and lKCs in the honey bee brain.