MALDI Imaging Analysis of Neuropeptides in Africanized Honeybee (Apis mellifera) Brain: Effect of Aggressiveness
MALDI Imaging Analysis of Neuropeptides in Africanized Honeybee (Apis mellifera) Brain: Effect of Aggressiveness
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DOI：
10.1021/acs.jproteome.8b00098
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发表时间：
2018-07-01
影响因子：
4.4
通讯作者：
Palma, Mario Sergio
中科院分区：
文献类型：
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作者：
Pratavieira, Marcel;da Silva Menegasso, Anally Ribeiro;Palma, Mario Sergio
蜜蜂的攻击性似乎受到多个基因的调控，在不同因素的影响下，如工蜂的聚乙二醇性、环境因素、对报警信息素的反应等，产生一系列行为反应。人们怀疑神经肽可能参与了攻击行为的调节。在攻击行为过程中，蜜蜂大脑中的Alatostatin和Tachykinin相关神经肽的作用尚不清楚，因此工蜂被刺激攻击和刺痛悬挂在蜂群前的皮革目标。收集攻击性个体，立即将其冷冻在液氮中；取出头部，在矢状面上切开。将脑片提交给MALDI光谱成像分析，本研究的结果报告了在攻击行为中，前体蛋白被加工成成熟形式的神经肽AmAST A(59-76)(AYTYVSEYKRLPVYNFGL-NH2)，AmAST A(69-76)(LPVYNFGL-NH2)，AmTRP(88-96)(APMGFQGMR-NH2)和AmTRP(254-262)(ARMGFHGMR-NH2)，它们显然在攻击行为中作用于蜜蜂大脑的不同神经髓，可能参与了这一复杂行为的不同方面的神经调制。这些结果通过对年轻的蜜蜂工蜂进行攻击性相关的行为分析得到了生物学上的验证，这些工蜂通过血肿接受了1 ng的AmAST A(69-76)或AmTRP(88-96)。这些年轻的工蜂在神经肽的存在下表现出了一系列完整的攻击性行为，证实了蜜蜂大脑中成熟的Amasts A和AmTRPs的存在与这种昆虫的攻击性有关的假说。
Aggressiveness in honeybees seems to be regulated by multiple genes, under the influence of different factors, such as polyethism of workers, environmental factors, and response to alarm pheromones, creating a series of behavioral responses. It is suspected that neuropeptides seem to be involved with the regulation of the aggressive behavior. The role of allatostatin and tachykinin-related neuropeptides in honeybee brain during the aggressive behavior is unknown, and thus worker honeybees were stimulated to attack and to sting leather targets hung in front of the colonies. The aggressive individuals were collected and immediately frozen in liquid nitrogen; the heads were removed and sliced at sagittal plan. The brain slices were submitted to MALDI spectral imaging analysis, and the results of the present study reported the processing of the precursors proteins into mature forms of the neuropeptides AmAST A (59-76) (AYTYVSEYKRLPVYNFGL-NH2), AmAST A (69-76) (LPVYNFGL-NH2), AmTRP (88-96) (APMGFQGMR-NH2), and AmTRP (254-262) (ARMGFHGMR-NH2), which apparently acted in different neuropils of the honeybee brain during the aggressive behavior, possibly taking part in the neuromodulation of different aspects of this complex behavior. These results were biologically validated by performing aggressiveness-related behavioral assays using young honeybee workers that received 1 ng of AmAST A (69-76) or AmTRP (88-96) via hemocele. The young workers that were not expected to be aggressive individuals presented a complete series of aggressive behaviors in the presence of the neuropeptides, corroborating the hypothesis that correlates the presence of mature AmASTs A and AmTRPs in the honeybee brain with the aggressiveness of this insect.