Department of Cell and Molecular Biology & Interdisciplinary Neurosciences Program, University of Rhode Island, Flagg Road, Kingston, RI, 02881, USA.
Institute of Applied Sciences and Intelligent Systems E. Caianiello, CNR, Via Campi Flegrei 34, 80078, Pozzuoli, Naples, Italy.
Cell Tissue Res. 2019 Aug;377(2):177-191. doi: 10.1007/s00441-019-03011-z. Epub 2019 Apr 11.
Previous biochemical studies have identified strychnine-sensitive glycine receptors in membrane preparations of Hydra vulgaris (Cnidaria: Hydrozoa). Electrophysiological and behavioral evidence has shown that these receptors play a role in modulating pacemaker activity and feeding behavior. Here, we present our genomic analysis that revealed hydra proteins having strong homology with the strychnine-binding region of the human receptor protein, GlyRα1. We further present immunocytochemical evidence for the specific labeling of cell and tissue preparations of hydra by a commercially available polyclonal anti-GlyRα1 antibody, selected through our genomic analysis. Tissue pieces and cell macerates from the upper and lower thirds of the body and ablated tentacles were double-labeled with this antibody and with an antibody specific for α-tubulin, to identify the glycine receptors and microtubules, respectively. Extensive receptor labeling was evident on the membranes, cell bodies and myonemes of endodermal and ectodermal epithelial cells, cell bodies and neurites of nerve cells, cnidocytes and interstitial cells. Labeling of the membranes of epithelial cells frequently corresponded to conspicuous varicosities (presumptive presynaptic sites) in the associated nerve net. Our findings support the idea that glycine receptors form an integral part of the nerve and effector systems that control hydra behavior.
先前的生化研究已经在水螅(腔肠动物门:水螅纲)的膜制剂中鉴定出士的宁敏感甘氨酸受体。电生理和行为证据表明,这些受体在调节起搏器活动和摄食行为方面发挥作用。在这里,我们展示了我们的基因组分析结果,该结果揭示了水螅蛋白与人类受体蛋白 GlyRα1 的士的宁结合区域具有很强的同源性。我们进一步通过免疫细胞化学证据证明了商业上可获得的抗 GlyRα1 抗体可以特异性标记水螅的细胞和组织制剂,该抗体是通过我们的基因组分析选择的。来自身体上下三分之一的组织片和细胞匀浆以及消融的触手用该抗体和针对α-微管蛋白的抗体进行双重标记,以分别鉴定甘氨酸受体和微管。在内胚层和外胚层上皮细胞的细胞膜、细胞体和肌动蛋白丝、神经细胞的细胞体和神经突、刺细胞和间质细胞上可见到广泛的受体标记。上皮细胞的膜标记经常与相关神经网中的明显膨体(假定的突触前部位)相对应。我们的发现支持这样一种观点,即甘氨酸受体是控制水螅行为的神经和效应系统的一个组成部分。