Analysis of the Differentiation of Kenyon Cell Subtypes Using Three Mushroom Body-Preferential Genes during Metamorphosis in the Honeybee (Apis mellifera L.).
Analysis of the Differentiation of Kenyon Cell Subtypes Using Three Mushroom Body-Preferential Genes during Metamorphosis in the Honeybee (Apis mellifera L.).
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DOI：
10.1371/journal.pone.0157841
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Kubo T
中科院分区：
文献类型：
--
作者：
Suenami S;Paul RK;Takeuchi H;Okude G;Fujiyuki T;Shirai K;Kubo T
成年蜜蜂（Apis mellifera L.）蘑菇体（MB，昆虫脑中的高级中心）包括四种亚型的内在神经元：I类大型、中型和小型凯尼恩细胞（分别为lKC、mKC和sKC）和II类KC。分析KC亚型在蜜蜂变态过程中的分化，对于深入了解KC亚型在蜜蜂行为中的作用具有重要意义。在本研究中，旨在确定KC亚型的标记基因，我们使用的cDNA微阵列，全面搜索在蜜蜂大脑中的MB优先方式表达的基因。在鉴定的18个基因中，我们进一步分析了其表达在MB中富集的三个基因：磷脂酶C β（PLCe）、突触结合蛋白14（Syt 14）和椎间盘大同源物5（dlg 5）。定量逆转录-聚合酶链反应分析显示，PLCe，Syt 14和dlg 5的表达更丰富的MBs比在其他脑区分别约31，6.8和5.6倍。原位杂交显示Syt 14和dlg 5的表达在lKCs中富集，但在mKCs和sKCs中不富集，而PLCe的表达在蜜蜂脑中的所有KC亚型（整个MB）中相似，这表明Syt 14和dlg 5以及PLCe分别可用作lKCs和所有KC亚型的标记基因。原位杂交显示，PLCe的表达已经检测到在第二类KC在幼虫五龄喂养阶段，表明PLCe的表达是一个共同的幼虫和成人MBs的特征。相反，Syt 14和dlg 5的表达在第三天蛹时变得可检测，表明Syt 14和dlg 5表达是晚期蛹和成虫MB的特征，并且在晚期蛹阶段建立了lKC特异性分子特征。
The adult honeybee (Apis mellifera L.) mushroom bodies (MBs, a higher center in the insect brain) comprise four subtypes of intrinsic neurons: the class-I large-, middle-, and small-type Kenyon cells (lKCs, mKCs, and sKCs, respectively), and class-II KCs. Analysis of the differentiation of KC subtypes during metamorphosis is important for the better understanding of the roles of KC subtypes related to the honeybee behaviors. In the present study, aiming at identifying marker genes for KC subtypes, we used a cDNA microarray to comprehensively search for genes expressed in an MB-preferential manner in the honeybee brain. Among the 18 genes identified, we further analyzed three genes whose expression was enriched in the MBs: phospholipase C epsilon (PLCe), synaptotagmin 14 (Syt14), and discs large homolog 5 (dlg5). Quantitative reverse transcription-polymerase chain reaction analysis revealed that expression of PLCe, Syt14, and dlg5 was more enriched in the MBs than in the other brain regions by approximately 31-, 6.8-, and 5.6-fold, respectively. In situ hybridization revealed that expression of both Syt14 and dlg5 was enriched in the lKCs but not in the mKCs and sKCs, whereas expression of PLCe was similar in all KC subtypes (the entire MBs) in the honeybee brain, suggesting that Syt14 and dlg5, and PLCe are available as marker genes for the lKCs, and all KC subtypes, respectively. In situ hybridization revealed that expression of PLCe is already detectable in the class-II KCs at the larval fifth instar feeding stage, indicating that PLCe expression is a characteristic common to the larval and adult MBs. In contrast, expression of both Syt14 and dlg5 became detectable at the day three pupa, indicating that Syt14 and dlg5 expressions are characteristic to the late pupal and adult MBs and the lKC specific molecular characteristics are established during the late pupal stages.