Mendis-Handagama S M, Aten R F, Watkins P A, Scallen T J, Berhman H R
Department of Animal Science, College of Veterinary Medicine, University of Tennessee, Knoxville 37996, USA.
Tissue Cell. 1995 Oct;27(5):483-90. doi: 10.1016/s0040-8166(05)80056-4.
In the present investigation, we have studied peroxisomes and sterol carrier protein-2 (SCP2) in control and luteinizing hormone stimulated rat luteal cells. Superovulated immature rats in mid-luteal phase (8 days after ovulation) were divided into two groups (n = 4/group) and treated with vehicle (0.2 ml saline), or luteinizing hormone (LH, 20 micrograms/rat). In this animal model, LH acutely stimulates steroidogenesis. Thirty minutes later, corpora lutea were fixed by whole body perfusion and processed for (1) electron microscopic immunocytochemistry to localize SCP2 via the protein A gold immunolabeling technique, and for (2) electron microscopic histochemistry to stain peroxisomal catalase via the alkaline 3,3'-diaminobenzidine tetrahydrochloride method. In the steroidogenic, mid-phase luteal cells of vehicle treated rats (controls), SCP2 was highly concentrated in peroxisomes and sparsely scattered on mitochondria, but no labeling was observed in lipid droplets. In the luteal cells of rats acutely stimulated with LH, peroxisomes immunolabeled for SCP2 were observed within the luteal cell lipid droplets and mitochondria, and in union with lipid droplets and mitochondria. Moreover, in contrast to control luteal cells, significant immunolabeling for SCP2 was detected within the lipid droplets and mitochondria in luteal cells of LH-treated rats. As SCP2 binds cholesterol to 1:1 molar ratio and is known to be involved in the intracellular movement of cholesterol, these findings suggest that peroxisomes and SCP2 may possibly be involved in delivering cholesterol from lipid droplets to the mitochondria when luteal cell steroidogenesis is acutely stimulated by LH.
在本研究中,我们研究了对照及促黄体生成素刺激的大鼠黄体细胞中的过氧化物酶体和固醇载体蛋白-2(SCP2)。处于黄体中期(排卵后8天)的超排卵未成熟大鼠被分为两组(每组n = 4),分别用溶剂(0.2 ml生理盐水)或促黄体生成素(LH,20微克/大鼠)处理。在这个动物模型中,LH急性刺激类固醇生成。30分钟后,通过全身灌注固定黄体,并进行如下处理:(1)通过蛋白A金免疫标记技术进行电子显微镜免疫细胞化学,以定位SCP2;(2)通过碱性3,3'-二氨基联苯胺四盐酸盐法进行电子显微镜组织化学,以对过氧化物酶体过氧化氢酶进行染色。在溶剂处理大鼠(对照组)的类固醇生成中期黄体细胞中,SCP2高度集中于过氧化物酶体,稀疏分布于线粒体,但在脂滴中未观察到标记。在LH急性刺激的大鼠黄体细胞中,在黄体细胞脂滴和线粒体内以及与脂滴和线粒体结合处观察到了SCP2免疫标记的过氧化物酶体。此外,与对照黄体细胞相比,在LH处理大鼠的黄体细胞脂滴和线粒体中检测到了显著的SCP2免疫标记。由于SCP2与胆固醇以1:1摩尔比结合,且已知参与胆固醇的细胞内转运,这些发现表明,当LH急性刺激黄体细胞类固醇生成时,过氧化物酶体和SCP2可能参与将胆固醇从脂滴转运至线粒体。