UCP induction on the inner cell and mitochondrial membrances of Xenopus oocyte injected with gene transcripts from brown adipocytes of cold acclimated rat

用酸性异硫氰酸胍酚氯仿(AGPC)法从冷习服和温习服大鼠棕色脂肪细胞中提纯RNA转录本。注入冷习服大鼠RNA转录本的卵母细胞，全细胞钳记录显示电压依赖性大电导，而注射温习服大鼠转录本的卵母细胞则无电导记录。光镜和电子显微镜观察显示，将冷驯化的大鼠棕色脂肪细胞的RNA转录产物注入卵母细胞后，在卵母细胞表面附近出现了新合成的内膜。用抗UCP1多克隆抗体进行免疫金电镜观察发现，在注射冷习服大鼠转录本的卵母细胞中，免疫金颗粒定位于新合成的线粒体膜的内膜上。

The RNA transcripts were purified from the brown adipocytes of cold and warm acclimated rats by the acid guanidium isothiocyanate phenol chloroform (AGPC) method. Whole cell clamp recordings of the oocyte which were injected with the RNA transcript purified from cold acclimated rats, revealed a voltage dependent large conductance, while no conductance was recorded on the oocyte injected with the transcript from warm acclimated rat. Light and electron microscopic observations showed the appearance of newly synthesized inner membrane near the surface of oocytes which were injected with RNA transcripts purified from cold acclimated rat brown adipocytes. Immuno-gold electron microscopy using anti UCP1 polyclonal antibody revealed the localization of immuno-gold particles on the inner membrane of mitochondria like structure of the newly synthesized membrane in oocytes injected with the cold acclimated rat transcript.