Characterization, localization and temporal expression of crustacean hyperglycemic hormone (CHH) in the behaviorally rhythmic peracarid crustaceans, Eurydice pulchra (Leach) and Talitrus saltator (Montagu)
Characterization, localization and temporal expression of crustacean hyperglycemic hormone (CHH) in the behaviorally rhythmic peracarid crustaceans, Eurydice pulchra (Leach) and Talitrus saltator (Montagu)
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DOI：
10.1016/j.ygcen.2016.07.024
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发表时间：
2016-10-01
影响因子：
2.7
通讯作者：
Wilcockson, David Charles
中科院分区：
文献类型：
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作者：
Hoelters, Laura;O'Grady, Joseph Francis;Wilcockson, David Charles
甲壳类高血糖激素（CHH）在十足甲壳类动物中被广泛研究，它具有多种作用，包括调节血糖水平。在生物钟的控制下，十足动物的高血糖似乎是暂时与活动周期相一致的。本研究采用基因克隆、原位杂交和免疫组织化学方法，对两种过壳类动物Eurydice pulchra和Talitrus saltator中CHH的特征和定位进行了研究。我们还利用这些物种强大的行为节律性来验证CHH mRNA表达与它们的周期（12.4 h）和昼夜（24 h）行为表型共振的假设。我们发现，这两个物种在大脑神经节中表达一个单一的CHH转录物，编码具有其他CHH所有预期的保守特征的肽。pulchra preproCHH是一个经修饰的73个氨基酸的肽，n端由一个短的18个氨基酸前体相关肽（CPRP）组成，而T. saltator原激素也经过修饰，但长度为72个氨基酸，残基为56个CPRP。两者的定位通过免疫组织化学映射到原大脑，轴突束通向窦腺和三头脑，与陆地等足类动物有着惊人的相似性。我们通过原位杂交证实了CHH免疫反应细胞的细胞位置。尽管这两个物种都表现出强大的活动节奏，但都没有表现出有节奏的转录活动，这表明CHH转录不太可能受时钟控制。这些数据有助于编制目前缺乏的非十足类物种的CHHs清单。(C) 2016 Elsevier Inc.版权所有。
Crustacean hyperglycemic hormone (CHH) has been extensively studied in decapod crustaceans where it is known to exert pleiotropic effects, including regulation of blood glucose levels. Hyperglycemia in decapods seems to be temporally gated to coincide with periods of activity, under circadian clock control. Here, we used gene cloning, in situ hybridization and immunohistochemistry to describe the characterization and localization of CHH in two peracarid crustaceans, Eurydice pulchra and Talitrus saltator. We also exploited the robust behavioral rhythmicity of these species to test the hypothesis that CHH mRNA expression would resonate with their circatidal (12.4 h) and circadian (24 h) behavioral phenotypes. We show that both species express a single CHH transcript in the cerebral ganglia, encoding peptides featuring all expected, conserved characteristics of other CHHs. E. pulchra preproCHH is an amidated 73 amino acid peptide N-terminally flanked by a short, 18 amino acid precursor related peptide (CPRP) whilst the T. saltator prohormone is also amidated but 72 amino acids in length and has a 56 residue CPRP. The localization of both was mapped by immunohistochemistry to the protocerebrum with axon tracts leading to the sinus gland and into the tritocerebrum, with striking similarities to terrestrial isopod species. We substantiated the cellular position of CHH immunoreactive cells by in situ hybridization. Although both species showed robust activity rhythms, neither exhibited rhythmic transcriptional activity indicating that CHH transcription is not likely to be under clock control. These data make a contribution to the inventory of CHHs that is currently lacking for non-decapod species. (C) 2016 Elsevier Inc. All rights reserved.