Acetylcholinesterase Regulates Skeletal In Ovo Development of Chicken Limbs by ACh-Dependent and -Independent Mechanisms.
Acetylcholinesterase Regulates Skeletal In Ovo Development of Chicken Limbs by ACh-Dependent and -Independent Mechanisms.
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DOI：
10.1371/journal.pone.0161675
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Layer PG
中科院分区：
文献类型：
--
作者：
Spieker J;Ackermann A;Salfelder A;Vogel-Höpker A;Layer PG
脊椎动物肢体的形成为分析非神经元胆碱能系统（NNCS）提供了一个很好的模型。在这里，我们首先分析了乙酰胆碱酯酶（AChE）的IHC和胆碱乙酰转移酶（ChAT）的ISH在发育中的胚胎鸡肢（阶段HH 17 -37）的表达。乙酰胆碱酯酶概述了骨骼的形成，在其远端最强，后来也标记了细胞死亡的区域。在发病时，AChE和ChAT升高的两个组织中心的肢体原基，顶端外胚层嵴（AER）和极化活动区（ZPA），分别。因此，ChAT表达后不久，乙酰胆碱酯酶，从而有力地支持了肢体形成的乙酰胆碱酯酶的主导作用。然后，我们进行了功能丧失的研究，通过单侧植入珠到鸡肢体原基，这是浸泡在胆碱能成分。在不同的时间后，软骨基质和矿化骨的形成分别用阿尔新蓝（AB）和茜素红（AR）染色。乙酰胆碱（ACh）和ChAT浸泡的珠子都加速了卵内骨形成。值得注意的是，通过BW 284 c51或通过单克隆抗体MAB 304抑制AChE延迟软骨形成。由于BChE的微珠抑制大多无效，因此表明在BW 284 c51和MAB 304抑制期间的ACh非依赖性作用，这可能是由于AChE的酶副活性。总之，鸡的骨骼发育受乙酰胆碱依赖性胆碱能系统的调节，但在一定程度上也由乙酰胆碱酯酶蛋白的乙酰胆碱独立方面。
Formation of the vertebrate limb presents an excellent model to analyze a non-neuronal cholinergic system (NNCS). Here, we first analyzed the expression of acetylcholinesterase (AChE) by IHC and of choline acetyltransferase (ChAT) by ISH in developing embryonic chicken limbs (stages HH17-37). AChE outlined formation of bones, being strongest at their distal tips, and later also marked areas of cell death. At onset, AChE and ChAT were elevated in two organizing centers of the limb anlage, the apical ectodermal ridge (AER) and zone of polarizing activity (ZPA), respectively. Thereby ChAT was expressed shortly after AChE, thus strongly supporting a leading role of AChE in limb formation. Then, we conducted loss-of-function studies via unilateral implantation of beads into chicken limb anlagen, which were soaked in cholinergic components. After varying periods, the formation of cartilage matrix and of mineralizing bones was followed by Alcian blue (AB) and Alizarin red (AR) stainings, respectively. Both acetylcholine (ACh)- and ChAT-soaked beads accelerated bone formation in ovo. Notably, inhibition of AChE by BW284c51, or by the monoclonal antibody MAB304 delayed cartilage formation. Since bead inhibition of BChE was mostly ineffective, an ACh-independent action during BW284c51 and MAB304 inhibition was indicated, which possibly could be due to an enzymatic side activity of AChE. In conclusion, skeletogenesis in chick is regulated by an ACh-dependent cholinergic system, but to some extent also by an ACh-independent aspect of the AChE protein.
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影响因子：
3
作者：
Farchi, Noa;Ofek, Keren;Soreq, Hermona
通讯作者：
Soreq, Hermona
影响因子：
3.6
作者：
LAYER, PG;ROMMEL, S;HENGSTENBERG, R
通讯作者：
HENGSTENBERG, R
影响因子：
4.8
作者：
Masha'our RS;Heinrich R;Garzozi HJ;Perlman I
通讯作者：
Perlman I
影响因子：
4.9
作者：
McAllen RM;Cook AD;Khiew HW;Martelli D;Hamilton JA
通讯作者：
Hamilton JA
影响因子：
6.7
作者：
Guo, Jie;Wang, Lue;Che, Xiaoxia
通讯作者：
Che, Xiaoxia