Amano Masafumi, Amiya Noriko, Yokoyama Takehiko
School of Marine Biosciences, Kitasato University, Sagamihara, Kanagawa 252-0373, Japan.
School of Marine Biosciences, Kitasato University, Sagamihara, Kanagawa 252-0373, Japan.
Acta Histochem. 2019 Feb;121(2):234-239. doi: 10.1016/j.acthis.2019.01.001. Epub 2019 Jan 5.
The gonadotropin-releasing hormone (GnRH) gene sequence has been identified in an annelid polychaete marine worm using continual genome sequencing. The distribution of GnRH immunoreactive (ir) cell bodies and fibers in the nerve ganglion of the clam worm Perinereis aibuhitensis (Polychaeta) was examined by immunohistochemistry using a newly produced rabbit polyclonal antibody raised against the marine worm GnRH (mwGnRH). The specificity of the antibody was confirmed by dot blot assay. The antibody cross-reacted with mwGnRH, but not with other forms of GnRH such as octopus GnRH, tunicate GnRH-I, II, owl limpet GnRH, and lamprey GnRH-II. In P. aibuhitensis, mwGnRH-ir cell bodies were detected in the nuclei 15-22, the caudal part of the cerebral ganglion. Furthermore, mwGnRH-ir fibers were mainly observed in the optic neuropil, but mwGnRH-ir fibers were also detected in the central neuropil region, the subpharyngeal ganglion, and the ventral nerve cord. These results indicate that mwGnRH is synthesized in the cerebral ganglion, is transported through the subpharyngeal ganglion and the ventral nerve cord, and functions either as a neurotransmitter or neuromodulator.
利用连续基因组测序技术,在一种环节多毛纲海洋蠕虫中鉴定出了促性腺激素释放激素(GnRH)基因序列。使用新制备的针对海洋蠕虫GnRH(mwGnRH)的兔多克隆抗体,通过免疫组织化学方法检测了沙蚕(Perinereis aibuhitensis,多毛纲)神经节中GnRH免疫反应性(ir)细胞体和纤维的分布。通过斑点印迹分析证实了该抗体的特异性。该抗体与mwGnRH发生交叉反应,但与其他形式的GnRH,如章鱼GnRH、被囊动物GnRH-I、II、帽贝GnRH和七鳃鳗GnRH-II不发生交叉反应。在沙蚕中,mwGnRH-ir细胞体在第15 - 22号细胞核中被检测到,位于脑神经节的尾部。此外,mwGnRH-ir纤维主要在视神经纤维网中观察到,但在中枢神经纤维网区域、咽下神经节和腹神经索中也检测到了mwGnRH-ir纤维。这些结果表明,mwGnRH在脑神经节中合成,通过咽下神经节和腹神经索运输,并作为神经递质或神经调节剂发挥作用。