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松鼠猴脑干核团中运动诱导的2-脱氧葡萄糖摄取变化:放射自显影和液体闪烁研究

Motion-induced alterations in 2-deoxyglucose uptake in brainstem nuclei of squirrel monkeys: autoradiographic and liquid scintillation studies.

作者信息

Brizzee K R, Dunlap W P

出版信息

Brain Behav Evol. 1983;23(1-2):14-25. doi: 10.1159/000121483.

Abstract

Each of 8 young adult female squirrel monkeys were injected via a femoral vein cannula with 167 microCi/100 g body weight of 2-(1,2-3H)-deoxy-D-glucose (2-DG) (New England Nuclear, 37.3 Ci/mmol) in 0.5 ml sterile saline. 4 additional female squirrel monkeys were injected in the same manner with 100 microCi/100 g body weight of the (3H)-2-DG. 2 h after this initial injection the original 8 animals were injected with 16.7 microCi/100 g body weight of 2-(1-14C)-deoxy-D-glucose (51.3 mCi/mmol) in 0.5 ml sterile saline. The 4 additional animals were injected with 25 microCi/100 g body weight of the (14C)-2-DG. Half of the animals each dose level were restrained in the upright position with Velcro straps and a nontraumatic moulded plastic head holder on a modified animal restraint board [Withrow and Devine, 1972] with the head in the sagittal plane but tilted forward about 20 degrees. They were then subjected to horizontal rotary motion at 25 rpm together with a vertical movement of 6 inches at 0.5 Hz for 1 h in a lighted room. The other half of the animals at each dose were restrained in the same manner and maintained in a quiescent state. At the end of this period each animal was anesthetized with ketamine, and the brain was quickly dissected out and frozen in isopentane cooled to -60 degrees C with dry ice. Transverse cryostat sections (-15 degrees C) of the brainstem were cut alternately at 200 and 20 microns from the nucleus gracilis caudally through the superior vestibular nucleus rostrally. Micropunch samples of the individual vestibular nuclei, and other brainstem nuclei and areas were obtained from the 200-microns sections with a modification of the method of Eik-Nes and Brizzee with a small stainless steel punch measuring 850 microns in diameter. The frozen punch samples were prepared for liquid scintillation counting (Beckman LS7500 system). Differential (3H) and (14C) counts (cpm) were made employing external standards. The 20-microns sections were prepared for 14C autoradiography by standard methods employing Kodak SB-5 X-ray film. The (14C) autoradiographs from the motion-stimulated animals revealed a selectively high uptake of 2-DG in all of the vestibular nuclei, the lateral cuneate nucleus and the lateral reticular nucleus as compared with the corresponding structures in the control animals.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

8只成年雌性松鼠猴通过股静脉插管,每100克体重注射167微居里的2 -(1,2 - 3H)-脱氧 - D -葡萄糖(2 - DG)(新英格兰核公司,37.3居里/毫摩尔),溶于0.5毫升无菌盐水中。另外4只雌性松鼠猴以同样方式每100克体重注射100微居里的(3H)-2 - DG。初次注射后2小时,最初的8只动物每100克体重注射16.7微居里的2 -(1 - 14C)-脱氧 - D -葡萄糖(51.3毫居里/毫摩尔),溶于0.5毫升无菌盐水中。另外4只动物每100克体重注射25微居里的(14C)-2 - DG。每个剂量水平的一半动物用维可牢尼龙搭扣带和改良动物约束板上的无创伤模制塑料头固定器固定在直立位置[威瑟罗和迪瓦恩,1972],头部位于矢状面但向前倾斜约20度。然后在有光照的房间里,它们以25转/分钟的速度进行水平旋转运动,并以0.5赫兹的频率进行6英寸的垂直运动,持续1小时。每个剂量的另一半动物以同样方式固定并保持静止状态。在此期间结束时,每只动物用氯胺酮麻醉,然后迅速取出大脑并在用干冰冷却至 - 60摄氏度的异戊烷中冷冻。脑干的横向低温恒温器切片(-15摄氏度)从尾侧的薄束核到吻侧的上前庭核,交替切成200微米和20微米厚。通过对艾克 - 内斯和布里兹方法的改进,用直径850微米的小不锈钢打孔器从200微米厚的切片中获取各个前庭核以及其他脑干核和区域的微打孔样本。冷冻的打孔样本准备用于液体闪烁计数(贝克曼LS7500系统)。采用外标进行(3H)和(14C)的差异计数(每分钟计数)。20微米厚的切片通过使用柯达SB - 5 X射线胶片的标准方法制备用于14C放射自显影。与对照动物的相应结构相比,运动刺激动物的(14C)放射自显影片显示所有前庭核、外侧楔束核和外侧网状核中2 - DG的摄取选择性地高。(摘要截断于400字)

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