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对十足目甲壳动物日本对虾(Marsupenaeus japonicus)中枢神经系统中一种促性腺激素释放激素(GnRH)样免疫反应性肽的生化分析和免疫组织化学检查。

Biochemical analysis and immunohistochemical examination of a GnRH-like immunoreactive peptide in the central nervous system of a decapod crustacean, the kuruma prawn (Marsupenaeus japonicus).

作者信息

Amano Masafumi, Okumura Takuji, Okubo Kataaki, Amiya Noriko, Takahashi Akiyoshi, Oka Yoshitaka

机构信息

School of Marine Bioscíences, Kitasato University, Ofunato, Iwate 022-0101, Japan.

出版信息

Zoolog Sci. 2009 Dec;26(12):840-5. doi: 10.2108/zsj.26.840.

Abstract

We examined whether a gonadotropin-releasing hormone (GnRH)-like peptide exists in the central nervous system (CNS) of the kuruma prawn, Marsupenaeus japonicus, by reverse-phase high performance liquid chromatography (rpHPLC) combined with time-resolved fluoroimmunoassay (TR-FIA) analysis and by immunohistochemistry. The displacement curve obtained for serially diluted extracts of the kuruma prawn brain paralleled the chicken GnRH-II (cGnRH-II) standard curve obtained by cGnRH-II TR-FIA using the anti-cGnRH-II antibody, which cross-reacts not only with cGnRH-II but also with lamprey GnRH-II (lGnRH-II) and octopus GnRH (octGnRH). Extracts of kuruma prawn brains and eyestalks showed a similar retention time to synthetic lGnRH-II and octGnRH in rpHPLC combined with TR-FIA analysis. Using this antibody, we detected GnRH-like-immunoreactive (ir) cell bodies in the anterior-most part of the supraesophageal ganglion (brain), the protocerebrum. Furthermore, GnRH-like-ir fibers were observed in the protocerebrum and deutocerebrum. In the eyestalk, GnRH-like-ir cell bodies were detected in the medulla interna, and GnRH-like-ir fibers were distributed in the medulla interna, medulla externa, and lamina ganglionalis. In the thoracic ganglion, GnRH-like-ir fibers, but not GnRH-like-ir cell bodies, were detected. No GnRH-like-ir cell bodies or fibers were detected in the abdominal ganglion or ovary. Thus, we have shown the existence and distribution of a GnRH-like peptide in the CNS of the kuruma prawn.

摘要

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,研究了日本对虾(Marsupenaeus japonicus)中枢神经系统(CNS)中是否存在促性腺激素释放激素(GnRH)样肽。日本对虾脑提取物系列稀释后的置换曲线与使用抗鸡GnRH-II(cGnRH-II)抗体通过cGnRH-II TR-FIA获得的鸡GnRH-II标准曲线平行,该抗体不仅与cGnRH-II交叉反应,还与七鳃鳗GnRH-II(lGnRH-II)和章鱼GnRH(octGnRH)交叉反应。在结合TR-FIA分析的rpHPLC中,日本对虾脑和眼柄提取物显示出与合成lGnRH-II和octGnRH相似的保留时间。使用该抗体,我们在食管上神经节(脑)最前端的原脑区域检测到了GnRH样免疫反应性(ir)细胞体。此外,在原脑和中脑检测到了GnRH样ir纤维。在眼柄中,在内髓部检测到GnRH样ir细胞体,GnRH样ir纤维分布在内髓部、外髓部和神经节层。在胸神经节中,检测到了GnRH样ir纤维,但未检测到GnRH样ir细胞体。在腹神经节或卵巢中未检测到GnRH样ir细胞体或纤维。因此,我们证明了日本对虾中枢神经系统中存在GnRH样肽并确定了其分布。

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