Schulz R
Cell Tissue Res. 1986;245(3):629-33. doi: 10.1007/BF00218565.
Antisera (AS) raised in rabbits against 17 beta-estradiol (E) and testosterone (T) were tested for their suitability to localize E and T on deparaffinized, rehydrated sections of preovulatory trout ovaries, using the unlabeled antibody technique. Conventional control experiments demonstrated the specificity of the staining reactions. Furthermore, no staining was observed after the removal of T-specific antibodies by affinity chromatography, or following gonadectomy when non-gonadal tissue sections of male trout were incubated with T-AS. Antiserum, raised against 11-oxotestosterone and devoid of antibodies cross-reacting with T, did not stain ovarian sections. The loci at which E and T are detected in the somatic compartment are consistent with the two-cell concept of estrogen synthesis, where aromatizable androgens are produced in the thecal/interstitial layer and serve as substrates for estrogen synthesis in granulosa cells. Both steroids were detected in yolk vesicles from the stage of endogenous vitellogenesis. T-AS showed affinity for nuclei of vitellogenic oocytes. Nucleoli were not stained.
使用未标记抗体技术,检测了在兔体内产生的针对17β-雌二醇(E)和睾酮(T)的抗血清,以确定其在排卵前鳟鱼卵巢脱蜡、复水切片上定位E和T的适用性。常规对照实验证明了染色反应的特异性。此外,通过亲和层析去除T特异性抗体后,或在雄性鳟鱼的非性腺组织切片与T抗血清孵育时进行性腺切除后,均未观察到染色。针对11-氧睾酮产生的抗血清,且不含与T交叉反应的抗体,未对卵巢切片进行染色。在体细胞区检测到E和T的位点与雌激素合成的双细胞概念一致,即在卵泡膜/间质层产生可芳香化的雄激素,并作为颗粒细胞中雌激素合成的底物。在内源性卵黄发生阶段的卵黄囊泡中检测到了这两种类固醇。T抗血清对卵黄生成卵母细胞的细胞核具有亲和力。核仁未被染色。