Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
J Mol Histol. 2010 Oct;41(4-5):215-24. doi: 10.1007/s10735-010-9281-7. Epub 2010 Aug 4.
Extracellular matrix in the ovarian follicle has been characterised for several mammalian species but there are no reports that describe the immunolocalisation of the extracellular matrix elements, matrix metalloproteinases, and its relation to plasma 17β estradiol levels and follicular apoptosis during the teleost's reproductive cycle. The present study used immunohistochemistry to characterise the distribution of laminin β2, collagen type IV, fibronectin and matrix metalloproteinases-9 (MMP-9). The TUNEL in situ technique was used to quantify apoptosis and indirect immunofluorimetric to determine plasma 17β estradiol levels. The TUNEL-positive reaction associated with morphological features exhibited follicular apoptosis. During postovulatory follicle involution, the drop in plasma 17β estradiol levels after spawning contributed to the intense apoptosis observed. By immunohistochemical analysis, laminin β2 and collagen type IV were identified as the major constituents of the basement membrane. The loss of integrity of the basement membrane occurred due to lyses of the major constituents, and coincides with increased follicular apoptosis. The integrity of the basement membrane is important for the survival of follicular cells. Furthermore, the MMP-9 results suggest that this enzyme is involved in final oocyte maturation and regression of postovulatory follicles. Fibronectin was observed on the surface of follicular cells of the postovulatory follicle in P. argenteus, this being important for maintaining normal cell adhesion to extracellular matrix. In conclusion, our results suggest that the structure and composition of the extracellular matrix, and plasma 17β estradiol levels related to apoptosis, play an important role during the follicular development and post-spawning involution in teleost fishes.
在几种哺乳动物物种中,已经对卵泡中的细胞外基质进行了特征描述,但没有报道描述细胞外基质成分、基质金属蛋白酶的免疫定位,以及它们与鱼类生殖周期中血浆 17β 雌二醇水平和卵泡凋亡的关系。本研究使用免疫组织化学技术来描述层粘连蛋白β2、IV 型胶原、纤维连接蛋白和基质金属蛋白酶-9(MMP-9)的分布。TUNEL 原位技术用于定量凋亡,间接免疫荧光法用于测定血浆 17β 雌二醇水平。与形态特征相关的 TUNEL 阳性反应表明卵泡发生凋亡。在排卵后卵泡退化期间,产卵后血浆 17β 雌二醇水平下降导致观察到强烈的凋亡。通过免疫组织化学分析,鉴定出层粘连蛋白β2 和 IV 型胶原作为基底膜的主要成分。基底膜的完整性丧失是由于主要成分的溶解引起的,并且与卵泡凋亡增加同时发生。基底膜的完整性对于卵泡细胞的存活很重要。此外,MMP-9 的结果表明,该酶参与了最终的卵母细胞成熟和排卵后卵泡的退化。在 P. argenteus 的排卵后卵泡中观察到纤维连接蛋白位于卵泡细胞表面,这对于维持正常细胞与细胞外基质的黏附很重要。总之,我们的结果表明,细胞外基质的结构和组成以及与凋亡相关的血浆 17β 雌二醇水平在鱼类卵泡发育和产卵后退化过程中起着重要作用。