Amano Masafumi, Moriyama Shunsuke, Okubo Kataaki, Amiya Noriko, Takahashi Akiyoshi, Oka Yoshitaka
School of Marine Biosciences, Kitasato University, Ofunato, Iwate, Japan.
Zoolog Sci. 2010 Aug;27(8):656-61. doi: 10.2108/zsj.27.656.
We examined whether gonadotropin-releasing hormone (GnRH)-like peptides are present in the neural ganglia of the gastropod Pacific abalone (Haliotis discus hannai) by reverse-phase high performance liquid chromatography (rpHPLC) combined with time-resolved fluoroimmunoassay (TR-FIA) analysis and by immunohistochemistry. Cerebral ganglion extracts showed a similar retention time to lamprey GnRH-II (lGnRH-II) in rpHPLC combined with TR-FIA analysis. GnRH-like-immunoreactive (ir) cell bodies (which reacted with a mouse monoclonal antibody raised against the common amino acid sequence of vertebrate GnRH) were detected in the peripheral region of the cerebral ganglion, and they were observed to send fibers into the neuropil. GnRH-like-ir fibers were also detected in the neuropil of the pedal ganglion, the visceral nerve, and the nerve originating from the pedal ganglion. Chicken GnRH-II (cGnRH-II)-like-ir fibers (which reacted with a rabbit polyclonal antibody raised against cGnRH-II) were also observed in the neuropil of the cerebral ganglion. GnRH-like-ir fibers and cGnRH-II-like-ir fibers were distinguishable in the neuropil of the cerebral ganglion by double-staining immunohistochemistry. These results suggest that multiple GnRH-like peptides exist in the neural ganglia of the Pacific abalone.
我们通过反相高效液相色谱(rpHPLC)结合时间分辨荧光免疫分析(TR-FIA)以及免疫组织化学方法,研究了腹足纲动物太平洋鲍鱼(Haliotis discus hannai)神经节中是否存在促性腺激素释放激素(GnRH)样肽。在rpHPLC结合TR-FIA分析中,脑神经节提取物显示出与七鳃鳗GnRH-II(lGnRH-II)相似的保留时间。在脑神经节的外周区域检测到GnRH样免疫反应性(ir)细胞体(与针对脊椎动物GnRH共同氨基酸序列产生的小鼠单克隆抗体发生反应),并且观察到它们发出纤维进入神经纤维网。在足神经节、内脏神经以及源自足神经节的神经的神经纤维网中也检测到了GnRH样ir纤维。在脑神经节的神经纤维网中还观察到了鸡GnRH-II(cGnRH-II)样ir纤维(与针对cGnRH-II产生的兔多克隆抗体发生反应)。通过双重染色免疫组织化学,在脑神经节的神经纤维网中可以区分GnRH样ir纤维和cGnRH-II样ir纤维。这些结果表明,太平洋鲍鱼的神经节中存在多种GnRH样肽。