摘要
用兔抗人ER-α和ER-β多克隆抗体对文昌鱼神经系统、轮器哈氏窝和性腺进行免疫细胞化学的定位研究。结果揭示幼年和成年两性不同发育时期文昌鱼在这些部位分布ER-α和ER-β蛋白。ER-α定位在端脑、中脑、后脑和神经管中大多数神经细胞核,少数在胞质及其突起和神经纤维,ER-β则定位在细胞质或细胞膜上,少数在核内。ER-α免疫阳性物质主要分布在哈氏窝下层的上皮细胞核,少数在上层细胞质,β受体则在上层细胞核。在性腺,ER-α分布在卵巢中卵原细胞和小生长期卵母细胞胞质与核仁,生发泡(核)显免疫阴性,在大生长期卵母细胞核膜和核仁的免疫阳性显著增强,成熟期则在卵细胞生发泡表达;ER-β免疫阳性物质分布在卵原细胞和早期卵母细胞质以及成熟卵细胞的卵被膜检测到,生发泡显免疫阴性。在精巢,这两种ER亚型均定位在精原细胞、初级与次级精母细胞和足细胞质,精子细胞在胞核,精子显免疫阴性。另外,双染结果还揭示ER-α和ER-β在上述部位多数共存于同一细胞,少数在不同细胞表达,且在细胞定位有不同。首次发现这两种雌激素受体亚型在文昌鱼有广泛分布,它们介导雌激素对文昌鱼神经内分泌组织的调节作用。α和β受体在靶细胞定位的不同,提示两者在介导雌激素信号路线和基因转录机制可能有不同生理作用。
Immunocytochemical localization were investigated in the nervous system, wheel organ, Hatschek's pit and gonads of amphioxus using polyclonal antibodies against estrogen receptor-α and β. The results revealed that ER-α and β protein distributed in the above regions in larvae and adult at different developmental stages of both sexes. A major ER-α were expressed within nucleus of nerve cells, a few expressed in the cytoplasm and process as well as fiber of nerve ecll in the forebram. (?)bram, hindbrain and nerve tube, while ER-β were detected in the cytoplasm or on the cellular niembiane, a few were within nucleus. ER-α immunopositive material distributed mainly in the nucleus of epithelial cells in the second layer of Hatschek's pit, a few were in the cytoplasm of upper layer epithelial cells, while ER-β distributed in the nucleic of upper layer. In gonads, ER-α were distributed in the cytoplasm and nucleolus of oogonia an: yte of small growth stage, germinal vesivie (nucleus) showed im-munonegative reaction. In the lange growths ge strong immunopositive reaction were showed in nuclear membrane and nucleolu of ooryte, and within nucleus of mature egg cell in the mature stage. ER-βimmunopositive mat(?) distributed in the cytoplasm of oogonia and early oocytes as well as egg envelope of mature egg cell, the germinal vesicle showed immunonegative reaction. In testis, both ER subtype were localized in the eyu plasm of spermatogonia, primary and second spermatocyte as well as Sertoli cell, and within nucleus of spermatid cell, while spermatozoa showed immunonegative reaction. On the other hand, the (?) of double staining revealed that ER-α and β numerously coexisted in a same cell, and fewly expressed in different cells in above regions. It is found for the first time that both estrogen receptor subtype, which mediated the regulate role of estrogen to neuroendocrine tissues in amphioxus, distributed extensively in amphiuoxus. The different localization of ER-α and β receptor in the target cells suggests t
出处
《实验生物学报》
CSCD
北大核心
2003年第5期368-374,共7页
Acta Biologiae Experimentalis Sinica
基金
国家自然科学基金(编号:39670104)
关键词
雌激素受体
文昌鱼
神经系统
哈氏窝
性腺
细胞定位
Amphioxus. Estrogen receptor (α and β). Nervous system. Hatschek's pit. Gonads
