Lucidi-Phillipi C A, Gage F H, Shults C W, Jones K R, Reichardt L F, Kang U J
Department of Neurosciences, University of California at San Diego, La Jolla 92093-0627, USA.
J Comp Neurol. 1995 Apr 10;354(3):361-76. doi: 10.1002/cne.903540306.
Local delivery of brain-derived neurotrophic factor (BDNF) by genetically modified cells provides the unique opportunity to examine the effects of BDNF on adult dopaminergic and cholinergic neurons in vivo. Primary rat fibroblasts were genetically engineered to produce BDNF. Conditioned media from BDNF-transduced fibroblasts supported embryonic chick dorsal root ganglion neurons as well as rat fetal mesencephalic neurons. BDNF-transduced fibroblasts grafted to the rat brain survived and showed continued mRNA production for at least 2 weeks. The effects of BDNF-transduced fibroblast grafts on the dopaminergic and cholinergic systems were then assessed. BDNF-transduced fibroblasts grafted into the normal intact substantia nigra induced sprouting of tyrosine hydroxylase- and neurofilament-immunoreactive fibers into the graft. Fibroblast grafts implanted into the normal intact striatum and midbrain as well as the 6-hydroxydopamine-lesioned brain did not induce sprouting of dopaminergic fibers; neither did they affect drug-induced rotational behavior. BDNF-transduced fibroblasts did, however, significantly increase the homovanillic acid/dopamine ratio when grafted into the normal midbrain. Following transection of the fimbriafornix, BDNF-transduced fibroblasts grafted into the septum were unable to rescue the septal cholinergic population, as did nerve growth factor-producing fibroblast grafts. Genetically modified fibroblast grafts may provide an effective, localized method of BDNF delivery in vivo to test biological effects of this factor on the central nervous system.
通过基因修饰细胞局部递送脑源性神经营养因子（BDNF）为在体内研究BDNF对成年多巴胺能和胆碱能神经元的作用提供了独特的机会。对原代大鼠成纤维细胞进行基因工程改造以产生BDNF。BDNF转导的成纤维细胞的条件培养基支持胚胎鸡背根神经节神经元以及大鼠胎儿中脑神经元。移植到大鼠脑中的BDNF转导的成纤维细胞存活下来，并显示出持续至少2周的mRNA产生。然后评估BDNF转导的成纤维细胞移植对多巴胺能和胆碱能系统的影响。移植到正常完整黑质中的BDNF转导的成纤维细胞诱导酪氨酸羟化酶和神经丝免疫反应性纤维向移植物中发芽。植入正常完整纹状体、中脑以及6-羟基多巴胺损伤脑内的成纤维细胞移植物未诱导多巴胺能纤维发芽；它们也不影响药物诱导的旋转行为。然而，当移植到正常中脑时，BDNF转导的成纤维细胞确实显著提高了高香草酸/多巴胺比率。在穹窿海马伞横断后，移植到隔区的BDNF转导的成纤维细胞无法像产生神经生长因子的成纤维细胞移植物那样挽救隔区胆碱能神经元群体。基因修饰的成纤维细胞移植物可能提供一种在体内有效、局部递送BDNF的方法，以测试该因子对中枢神经系统的生物学作用。
