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曼氏无针乌贼(Sepiella maindroni)脑结构及促性腺激素释放激素免疫组织化学定位研究

The Brain Structure and Gonadotropin-releasing Hormone Location by Immunohistochemistry in Sepiella Maindroni

【作者】 于新秀

【导师】 吴常文;

【作者基本信息】 浙江海洋学院 , 海洋生物学, 2011, 硕士

【摘要】 本文以性成熟的曼氏无针乌贼为研究对象,围绕头足类生殖调控轴亚脑脚叶/嗅叶后方——视腺——性腺轴,做了三个方面的研究:采用组织切片法对曼氏无针乌贼的中枢神经系统的神经叶结构组成进行了显微结构观察;采用组织切片和电镜技术对曼氏无针乌贼脑中的内分泌器官视腺做了显微结构和超显微结构的观察;采用免疫组织化学的方法对脑及生殖腺内的促性腺激素释放激素进行定位检测。结果表明:1.曼氏无针乌贼脑结构:成熟乌贼的食道穿过脑,以此可分为三部分,即食道上神经团、食道下神经团以及位于脑两侧两个发达的视叶,各个神经团又可以细分为不同的神经叶。食道上神经团可以分为垂直叶复合物、前后基叶、视神经束区、前连合叶以及脑亚脚叶,其中视神经束区为其调控生殖的区域。食道下神经团按相对位置分为前区、中区和后区,各区又可以细分为不同的神经叶。2.曼氏无针乌贼脑中内分泌器官——视腺显微结构和超显微结构观察结果表明视腺的分泌细胞具有分泌蛋白类类激素的特征。视腺位于视神经束上,靠近视叶,恰好位于嗅叶和背外侧叶之间。外周有一层结缔组织包裹,内部数量最多的细胞为分泌细胞,细胞核较大,直径约10μm,仅含有很少量的细胞质。另含有支持细胞、神经细胞等,而且支持细胞的细胞核致密较小。细胞间的结缔组织丰富,神经纤维穿梭其中。透射电镜观察发现分泌细胞的细胞核中染色质基本为常染色质,仅有少量的异染色质。细胞质中含有丰富的粗面内质网,上面可以观察到高电子密度的颗粒,高尔基体很活跃,形成高尔基体复合物,分泌有膜包被的小泡。另外在分泌细胞的周围还有血管分布,这可能便于激素类物质的运输。此研究将为进一步研究曼氏无针乌贼的视腺生理结构以及发育提供了细胞形态学基础。3.曼氏无针乌贼体内促性腺激素释放激素的定位与分布:采用免疫组织化学的方法,用哺乳动物的促性腺激素释放激素mGnRH抗体对曼氏无针乌贼的中枢神经系统进行检测定位,显微镜观察结果表明GnRH主要存在于视叶、背基叶、垂直叶、嗅叶等部位,这些神经叶都位于食道上神经团,而且围绕在视腺周围。另外在雌性的缠卵腺以及雄性的精巢中也检测到了它的存在。说明曼氏无针乌贼中的GnRH可以直接作用于性腺。

【Abstract】 According to the reproductive regulation axis subpeduncle lobe/olfactory lobe—optic gland—gonads, the research on mature Sepiella maindroni consisted of three parts:firstly, the brain structure of S. maindroni was performed by paraffin section observations; secondly, the structure and ultrastructure of the optic gland, an endocrine organ, were obtained; thirdly, whether a gonadotropin-releasing hormone (GnRH)-like peptide existed in the brain and gonads of S. maindroni was examined by performing an immunohistochemistry method.1. The morphology structure of mature S. maindroni brain was observed originally. The brain was divided into three parts: supraoesophageal mass, suboesophageal mass and two advanced optic lobes laterally, referring to the esophagus. Supraoesophageal mass included vertical lobe complex, anterior and posterior basal lobes, optic tract region, precommissural lobe and subpeduncle lobe. Optic tract region was involved in regulation of reproduction. Suboesophageal mass could be devided into three regions and each region could be subdivided into different lobes.2. The microstructure and ultrastructure of the optic gland revealed that the secretoy cells produced protein hormone. The gland covered by a thin connective layer was sited on each optic tract and very close to the optic lobe. It was just between the olfactory lobe and the dorsal lateral lobe. There existed abundant secretory cells with a diameter of nucleus 10μm approximately. The ultrastructure of the secretory cells was examined using Transmission Electron Microscope (TEM). There were numerous rough endoplasmic reticula, Golgi apparatus and electron dense granules in the cells. Blood vessels around secretory cells made it possible for hormones to transport. These results will provide cytomorphology basis for the research on physiology and development of optic glands in S. maindroni.3. The localization and distribution of gonadotropin-releasing hormone(GnRH) in S. maindroni were showed through immunohistochemistry. Location of GnRH was detected by mammal GnRH as a primary antibody. Observations by microscope were indicated that the optic lobe, basal lateral lobe,vertical lobe and olfactory lobe were the sites where GnRH existed. All of the lobes were in supraoesophageal mass and surrounded the optic gland. The presence of GnRH was also detected in nidamental gland and testis. The result suggested that GnRH ccould act on the gonads in S. maindroni directly.

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