Fujimori K, Rodgers K E, Nakamura R M, Katt E, Yanagihara D L, diZerega G S
Department of Obstetrics and Gynecology, University of Southern California School of Medicine, Los Angeles 90033.
J Histochem Cytochem. 1988 Jun;36(6):589-95. doi: 10.1177/36.6.3367045.
The granulosa cell secretes a protein (follicle regulatory protein: FRP) that affects the responsiveness of other follicles to gonadotropin stimulation. This protein was purified, partially characterized, and rabbit antisera as well as monoclonal antibodies were prepared against FRP. Fixed sections of porcine ovaries were prepared on slides and then incubated with the monoclonal antibody or polyclonal antisera and then incubated with either biotinylated mouse IgM or rabbit IgG antisera, respectively. These sections were then incubated with avidin conjugated to horseradish peroxidase, followed by substrate. Staining with both the monoclonal antibody and the antisera was present in the cytoplasm of granulosa cells of small- or medium-sized antral follicles. Staining distribution was localized preferentially to cells near the basal lamina; the antral granulosa cells of viable follicles did not stain. Neither primordial follicles nor pre-antral follicles (less than 300 microns in diameter) showed any positive staining. Thecal cells were not stained in follicles less than 5 mm in diameter, whereas some large follicles (greater than 5 mm) contained staining in the theca. In the latter, specific granulosa staining was only weakly positive with the polyclonal antibody and negative with the monoclonal antibody. Atretic follicles contained significant staining of all epithelial cells adjacent to the basal lamina by both the monoclonal and polyclonal antibody preparations. Staining of the luteal ovary by the monoclonal antibody was limited to the large luteal cells. These findings suggest that FRP is produced by the granulosa cells of porcine follicles at the stage of maturation corresponding to 0.5 mm in diameter. As the viable follicle increases in size, production of FRP in the granulosa is reduced below the detectable level when the follicle exceeds 5 mm in diameter. The main source of FRP during the luteal phase is the large cell of the corpus luteum.
颗粒细胞分泌一种蛋白质（卵泡调节蛋白：FRP），该蛋白会影响其他卵泡对促性腺激素刺激的反应性。对这种蛋白质进行了纯化、部分特性鉴定，并制备了针对FRP的兔抗血清以及单克隆抗体。将猪卵巢的固定切片制备在载玻片上，然后分别与单克隆抗体或多克隆抗血清孵育，接着再分别与生物素化的小鼠IgM或兔IgG抗血清孵育。随后将这些切片与与辣根过氧化物酶偶联的抗生物素蛋白孵育，再加入底物。使用单克隆抗体和抗血清染色均出现在中小卵泡腔颗粒细胞的细胞质中。染色分布优先定位于靠近基膜的细胞；存活卵泡的腔颗粒细胞未染色。原始卵泡和直径小于300微米的窦前卵泡均未显示任何阳性染色。直径小于5毫米的卵泡中，膜细胞未染色，而一些直径大于5毫米的大卵泡的膜中有染色。在后者中，用多克隆抗体检测时，颗粒细胞的特异性染色仅呈弱阳性，而用单克隆抗体检测则为阴性。闭锁卵泡中，单克隆和多克隆抗体制备物对所有与基膜相邻的上皮细胞均有明显染色。单克隆抗体对黄体卵巢的染色仅限于大黄体细胞。这些发现表明，FRP是由猪卵泡直径达0.5毫米时的颗粒细胞产生的。随着存活卵泡体积增大，当卵泡直径超过5毫米时，颗粒细胞中FRP的产生会降至可检测水平以下。黄体期FRP的主要来源是黄体的大细胞。
