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

采用异硫氰酸-苯酚-氯仿（AGPC）法从冷热驯化大鼠棕色脂肪细胞中纯化RNA转录本。转染冷驯化大鼠RNA转录物的卵母细胞在全细胞钳夹下显示出电压依赖性的大电导，而转染热驯化大鼠RNA转录物的卵母细胞则无电导记录。用冷驯化大鼠棕色脂肪细胞纯化的RNA转录本注射卵母细胞后，在光镜和电镜下观察到卵母细胞表面新合成的内膜。使用抗UCP1多克隆抗体进行免疫金电镜观察，发现在冷驯化大鼠转录本注入卵母细胞后，新合成的膜的线粒体样结构的内膜上定位了免疫金颗粒。

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.