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、热适应大鼠棕色脂肪细胞中提取RNA转录物。将冷适应大鼠的RNA转录物注射到卵母细胞中，用全细胞钳记录显示了电压依赖性的大电导，而将热适应大鼠的RNA转录物注射到卵母细胞中则没有电导记录。光镜和电镜观察显示，新合成的内膜表面附近的卵母细胞注射从冷驯化大鼠棕色脂肪细胞纯化的RNA转录本的外观。免疫金电子显微镜使用抗UCP1多克隆抗体显示定位的免疫金颗粒的线粒体样结构的新合成的膜在卵母细胞注射冷驯化大鼠转录的内膜。

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.