Expression and release patterns of neuropeptides during embryonic development and hatching of the green shore crab, Carcinus maenas
Expression and release patterns of neuropeptides during embryonic development and hatching of the green shore crab, Carcinus maenas
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DOI：
10.1242/dev.01312
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发表时间：
2004-10-01
期刊：
影响因子：
4.6
通讯作者：
Webster, SG
中科院分区：
文献类型：
--
作者：
Chung, JS;Webster, SG
甲壳类动物的蜕皮至少由三种神经肽控制：蜕皮抑制激素（MIH），抑制蜕皮类固醇的合成；甲壳动物高血糖激素（CHH），它不仅影响蜕皮类固醇的合成，而且影响换羽过程中的水吸收；和甲壳动物心脏活性肽（CLAP），它参与定型蜕皮行为。在胚胎发育过程中，蜕皮发生在卵中，但关于这一时期或孵化期间这些神经肽的发育表达的信息很少——这一事件类似于昆虫的羽化。为了解决这个问题，我们通过定量 RTPCR 以及发育肽表达研究 [共聚焦免疫细胞化学 (ICC) 和放射免疫分析 (RIA)] 确定了 MIH 和 CHH mRNA 的表达谱。与成虫蜕皮相比，在幼虫孵化期间观察到与 CHH 和 CCAP 神经肽激增相关的可能同源事件，并且 MIH 和 CHH mRNA 的细胞特异性拷贝浓度与胚胎发育后期成虫的相同。我们测量了发育过程中两种神经肽（红色素浓缩激素 RPCH）和色素分散激素（PDH）的平行 mRNA 和肽表达，因为它们具有神经调节剂和经典神经激素的作用。对于MIH和CHH，基因表达与肽表达一致，但发现了CHH表达的新位点（腹部周围神经元），其表达和释放模式可能与幼虫羽化和蛋壳破裂和孵化所需的水吸收有关。对于 RPCH 和 PDH，基因转录和肽表达不一致。母体衍生的（非翻译的）PDH mRNA 对胚胎有显着贡献，对于 RPCH，胚胎发生后期的高水平 mRNA 和肽表达与长期被忽视但可能重要的释放位点 - 神秘的连合后器官 - 相关，这是胚胎发生晚期表达 RPCH 的最突出的结构。
Crustacean ecdysis is controlled by at least three neuropeptides: moult-inhibiting hormone (MIH), which represses ecdysteroid synthesis; crustacean hyperglycaemic hormone (CHH), which not only influences ecdysteroid synthesis but also water uptake during moulting; and crustacean cardioactive peptide (CLAP), which is involved in stereotyped ecdysis behaviour. During embryonic development, moulting takes place in the egg, but there is little information regarding developmental expression of these neuropeptides during this period or during hatching - an event that is analogous to eclosion in insects. To address this problem, we determined expression profiles of MIH and CHH mRNA by quantitative RTPCR, together with developmental peptide expression studies [confocal immunocytochemistry (ICC) and radioimmunoassay (RIA)]. Likely homologous events relating to neuropeptide surges of both CHH and CCAP were seen during larval hatching, when compared to the adult moult, and cell-specific copy concentration of both MIH and CHH mRNAs was identical to that of the adult during late embryonic development. We measured parallel mRNA and peptide expression of two neuropeptides (red pigment-concentrating hormone RPCH) and pigment-dispersing hormone (PDH) during development, as these have roles as neuromodulators and as classical neurohormonal roles. For MIH and CHH, gene expression was in accordance with peptide expression, but novel sites of CHH expression were found (abdominal peripheral neurones), the expression and release patterns of which may be related to larval eclosion and water uptake necessary for eggshell rupture and hatching. For RPCH and PDH, gene transcription and peptide expression were not in accordance. A significant contribution of maternally derived (non-translated) PDH mRNA to the embryo was seen, and for RPCH, high-level mRNA and peptide expression during late embryogenesis is related to a long ignored, but potentially important release site - the enigmatic post-commissural organs - which are the most prominent structures expressing RPCH during late embryogenesis.