García-Ayala A, García-Hernández M P, Quesada J A, Agulleiro B
Department of Cell Biology, Faculty of Biology, University of Murcia, Spain.
Anat Rec. 1998 Apr;250(4):448-58. doi: 10.1002/(SICI)1097-0185(199804)250:4<448::AID-AR8>3.0.CO;2-T.
Gonadotropins GTH I and GTH II from the pituitary of Mediterranean (M.) yellowtail (Seriola dumerilii) were isolated and characterized, and antisera to the whole GTH II molecule (anti-My alpha,betaGTH II) and to its beta-subunit (anti-My betaGTH II) were obtained. At the light microscopic level, anti-My alpha,betaGTH II reacted with My betaGTH II-immunoreactive cells (GTH II cells), thyroid-stimulating hormone (TSH) cells, and a third cell population, which could have been GTH I cells. The aim of this study was the ultrastructural characterization of GTH and TSH cells in M. yellowtail using the immunogold method in order to provide a basis for future research into reproduction of this species.
Pituitaries from mature male and female specimens reared in captivity were dissected out and processed for electron microscopy. The immunogold method was carried out by using anti-My alpha,betaGTH II, anti-My alpha,betaGTH II preabsorbed with the alpha subunit of the M. yellowtail GTH (My alphaGTH-subunit), anti-My betaGTH II, anti-human (h) alpha,betaTSH, and anti-h betaTSH sera to reveal gonadotropic and thyrotropic cells.
M. yellowtail gonadotropic cells were very heterogeneous with regard to their size, shape, and ultrastructural features. Cells were found with numerous, round, variably electron-dense, secretory granules and globules; others were found with their cytoplasm occupied mostly by dilated cisternae of rough endoplasmic reticulum (RER) and scarce secretory granules; and other intermediate cell forms were found that showed varying proportions of secretory granules and dilated RER. The secretory granules and globules were immunogold labeled with anti-My alpha,betaGTH II, and the reaction was weaker in the latter. A similar immunogold-labeling pattern was found with anti-My betaGTH II and with anti-My alpha,betaGTH II preabsorbed with the My alphaGTH-subunit, although some cells that showed the same ultrastructural features described above were not immunogold labeled and could have been GTH I cells. Thyrotropic cells had small, round, secretory granules of medium or high electron density that were immunogold labeled with anti-My alpha,betaGTH II, anti-h alpha,betaTSH, and anti-h betaTSH sera, but not with anti-My betaGTH II or anti-My alpha,betaGTH II serum preabsorbed with the My alphaGTH-subunit. All of the cell forms described for gonadotropes and thyrotropes were also found in a state of involution.
Gonadotropes that are of a single morphological type but that vary in ultrastructure are present in the pituitary of captive M. yellowtail. GTH II- and putative GTH I-producing cells were distinguishable from one another and from TSH cells by their different reactions to anti-My alpha,betaGTH II, anti-My betaGTH II, and anti-My alpha,betaGTH II preabsorbed with the My alphaGTH-subunit.
从地中海黄尾鰤(Seriola dumerilii)垂体中分离并鉴定了促性腺激素GTH I和GTH II,并获得了针对整个GTH II分子(抗Myα,βGTH II)及其β亚基(抗MyβGTH II)的抗血清。在光镜水平上,抗Myα,βGTH II与MyβGTH II免疫反应性细胞(GTH II细胞)、促甲状腺激素(TSH)细胞以及第三种细胞群体发生反应,第三种细胞群体可能是GTH I细胞。本研究的目的是利用免疫金法对地中海黄尾鰤的GTH和TSH细胞进行超微结构表征,为该物种繁殖的未来研究提供基础。
解剖圈养的成熟雄性和雌性标本的垂体并进行电子显微镜处理。使用抗Myα,βGTH II、用地中海黄尾鰤GTH的α亚基(MyαGTH亚基)预吸收的抗Myα,βGTH II、抗MyβGTH II、抗人(h)α,βTSH和抗hβTSH血清进行免疫金法,以揭示促性腺细胞和促甲状腺细胞。
地中海黄尾鰤的促性腺细胞在大小、形状和超微结构特征方面非常异质。发现一些细胞含有大量圆形、电子密度可变的分泌颗粒和球状体;另一些细胞的细胞质主要被粗面内质网(RER)的扩张池占据,分泌颗粒稀少;还发现了其他中间细胞形式,其分泌颗粒和扩张的RER比例各不相同。分泌颗粒和球状体用抗Myα,βGTH II进行免疫金标记,后者的反应较弱。用抗MyβGTH II和用MyαGTH亚基预吸收的抗Myα,βGTH II也发现了类似的免疫金标记模式,尽管一些具有上述相同超微结构特征的细胞没有被免疫金标记,可能是GTH I细胞。促甲状腺细胞具有小的、圆形的、中等或高电子密度的分泌颗粒,用抗Myα,βGTH II、抗hα,βTSH和抗hβTSH血清进行免疫金标记,但用抗MyβGTH II或用MyαGTH亚基预吸收的抗Myα,βGTH II血清则没有标记。所描述的促性腺细胞和促甲状腺细胞的所有细胞形式也处于退化状态。
圈养的地中海黄尾鰤垂体中存在单一形态类型但超微结构不同的促性腺细胞。产生GTH II和假定的GTH I的细胞通过它们对抗Myα,βGTH II、抗MyβGTH II和用MyαGTH亚基预吸收的抗Myα,βGTH II的不同反应,彼此之间以及与TSH细胞区分开来。