EFFECTS OF CELL CULTURE TRANSPLANTS ON CNS REGENERATION

最近的研究引起了相当大的兴趣，这些研究已经 利用移植促进损伤后轴突的再生 成年哺乳动物中枢神经系统(CNS)。然而，目前几乎没有 已知正向轴突生长促进的细胞来源 似乎存在于周围神经或胚胎中枢神经系统的因子 组织。因此，拟议研究的长期目标是 鉴定和表征促进轴突再生的因素 成年哺乳动物中枢神经系统损伤的神经元。实验范式以 拟用于研究的是移植的各种 细胞和无细胞底物进入双侧头腔 所有的轴突连接都被切断了隔和 海马体。将使用四种实验性移植准备：1) 组织培养制备特定的非神经元三叉神经节或中枢神经系统细胞，2) 对照无细胞底物，3)完整的CNS或PNS神经，4) 细胞外基质分子。这些实验的具体目的是 重点确定移植环境对：1) 轴突再生速度，2)产生轴突的神经元类型，3) 促进轴突再生的特定细胞和/或分子，以及4) 再生轴突与细胞的形态关系 移植的地形。 几个光镜神经解剖学程序将被用于 识别和追踪移植和宿主之间的轴突连接 中枢神经系统。这将包括逆行追踪程序(荧光染料 和辣根过氧化物酶)、顺行追踪技术 (放射自显影、辣根过氧化物酶、神经纤维和髓鞘 染色)，用于识别特定轴突类型的组织化学方案 (儿茶酚胺组织荧光和乙酰胆碱酯酶组织化学)， 以及用于识别选定细胞的免疫组织化学染色技术 成分(层粘连蛋白、纤维连接蛋白、胶质纤维酸性蛋白、髓鞘 碱性蛋白质、5-羟色胺和C-4)。另外，那些实验 分析移植体内和移植宿主处的细胞事件 INTERFACE将使用透射电子显微镜。 拟议中的研究结果将为我们提供重要的见解 促进轴突再生的细胞和细胞外因子 在成年哺乳动物中枢神经系统中。这些轴突生长的特征 促进因素应被证明有利于开发治疗 治疗男性创伤性脑和脊髓损伤。

Considerable interest has been generated by recent studies which have utilized transplants to facilitate the regeneration of axons in the injured adult mammalian central nervous system (CNS). However, little is currently known about the cellular sources of the positive axonal growth promoting factors that appear to be present in peripheral nerve or embryonic CNS tissue. Thus, the long-term objectives of the proposed research are to identify and characterize factors which promote axonal regeneration from injured neurons in the adult mammalian CNS. The experimental paradigm to be used for the proposed studies is the transplantation of a variety of cellular and acellular substrates into bilateral intracephalic cavities in which all axonal connections are severed between the septum and hippocampus. Four experimental transplant preparations will be used: 1) specific non-neuronal PNS or CNS cells prepared in tissue culture, 2) control acellular substrates, 3) intact CNS or PNS nerves, and 4) extracellular matrix molecules. The specific aims of these experiments are focused to determine the effects of the transplant environments on: 1) the rate of axonal regrowth, 2) the types of neurons generating axons, 3) specific cells and/or molecules which promote axonal regeneration, and 4) the morphological relationship between regenerating axons and the cellular terrain of the transplant. Several light microscopic neuroanatomical procedures will be utilized for identifying and tracing axonal connections between the transplant and host CNS. These will include retrograde tracing procedures (fluorescent dyes and horseradish peroxidase), anterograde tracing techniques (autoradiography, horseradish peroxidase, neurofibrillar and myelin stains), histochemical protocols for identifying specific axon types (catecholamine histofluorescence and acetylcholinesterase histochemistry), and immunohistochemical staining techniques for identifying selected cell components (laminin, fibronectin, glial fibrillary acidic protein, myelin basic protein, serotonin and C-4). In addition, those experiments which analyze cellular events in the transplant and at the transplant-host interface will utilize transmission electron microscopy. The results from the proposed research will provide important insight into the cellular and extracellular factors that can promote axonal regeneration in the adult mammalian CNS. The characterization of these axonal growth promoting factors should prove beneficial for developing therapies for treating traumatic brain and spinal cord injuries in man.