Pulford B E, Mihajlov A R, Nornes H O, Whalen L R
Department of Anatomy and Neurobiology, Colorado State University, Ft. Collins 80523.
J Neural Transplant Plast. 1994 Apr-Jun;5(2):89-102. doi: 10.1155/NP.1994.89.
The effects of implantation of cultured adrenal medullary cells on the recovery of neurotransmitter specific reflex activity were studied in the rat spinal cord using electrophysiological testing methods. Cell suspensions of cultured neonatal adrenal medullary chromaffin (AM) cells (which produce catecholamines), or Schwann (Sc) cells (controls) were implanted into the lumbar region of the spinal cord 2 weeks after catecholamine (CA) denervation by intracisternal injection of 6-hydroxydopamine (6-OHDA). All cells were taken from 7 day neonates and cultured for 10 days in the presence of nerve growth factor (NGF). Three months after implantation, the extent of implant-associated recovery of reflex activity was determined by measuring electromyogram (EMG) activity and force associated with the long latency component of the hindlimb withdrawal reflex (which is CA modulated). After the electrophysiological testing, rats were anesthetized, and the spinal cords were rapidly removed and frozen. Spinal cords were sectioned longitudinally, and implanted cells were visualized using glyoxylic acid techniques. Labelled sections were examined to determine cell survival. Results indicate that 1) chromaffin cells survive for 3 months in the segments of the cord into which they have been implanted and 2) rats implanted with AM cells have significantly more forceful withdrawal reflexes than those that received Sc cells or received no implant after lesioning.
采用电生理测试方法,在大鼠脊髓中研究了植入培养的肾上腺髓质细胞对神经递质特异性反射活动恢复的影响。通过脑池内注射6-羟基多巴胺(6-OHDA)使儿茶酚胺(CA)去神经支配2周后,将培养的新生肾上腺髓质嗜铬(AM)细胞(产生儿茶酚胺)或雪旺氏(Sc)细胞(对照)的细胞悬液植入脊髓腰段。所有细胞均取自7日龄新生儿,并在神经生长因子(NGF)存在的情况下培养10天。植入后三个月,通过测量与后肢退缩反射的长潜伏期成分相关的肌电图(EMG)活动和力量(该反射受CA调节)来确定植入相关的反射活动恢复程度。电生理测试后,将大鼠麻醉,迅速取出脊髓并冷冻。将脊髓纵向切片,使用乙醛酸技术观察植入的细胞。检查标记的切片以确定细胞存活情况。结果表明:1)嗜铬细胞在其植入的脊髓节段中存活3个月;2)植入AM细胞的大鼠比接受Sc细胞或损伤后未植入的大鼠具有更有力的退缩反射。