Otto D, Unsicker K
Department of Anatomy and Cell Biology, University of Heidelberg, Germany.
J Neurosci Res. 1993 Mar 1;34(4):382-93. doi: 10.1002/jnr.490340403.
The protective role of basic fibroblast growth factor (FGF-2) for 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)- and methylpyridiniumion (MPP+)-lesioned dopaminergic (DAergic) nigrostriatal neurons was studied, using dissociated cell cultures of embryonic day (E) 14 rat mesencephalon. Cells were grown in different culture media and received FGF-2 (5 ng/ml) and/or the toxins (5 microM) at various schedules, but were consistently allowed to differentiate for 3 days prior to becoming exposed to the toxin. Survival of tyrosine hydroxylase (TH)-immunoreactive cells at 7 days was only markedly impaired by MPTP, if horse serum (HS) or bovine serum albumin (BSA) were omitted from the culture medium. FGF-2 increased the number of TH-immunoreactive cells, and this increase was not diminished by MPTP under any culture condition. Uptake of 3H-DA was significantly reduced by MPTP in HS- and BSA-containing, but not in protein-less cultures. A protective effect by FGF-2 was only seen in the presence of BSA. MPP+ caused a more pronounced reduction in 3H-DA uptake than MPTP, and this effect was partially reversed by the addition of FGF-2, unless cultures contained HS. Neurofilament protein (NF), and indirect measure for the total number of neurons present in the cultures, was not significantly reduced by MPTP or MPP+ corroborating the specificity of the toxin for DAergic neurons, which constitute only a minor fraction in these cultures. In line with the wide spectrum of target neurons of FGF-2, this factor significantly increased NF contents under any culture condition. Quantification of the amounts of glial fibrillary acidic protein (GFAP) revealed stimulatory effects of FGF-2 (2.5- to 4-fold) and at least 10-fold higher levels in the presence as compared to the absence of HS. These data show that FGF-2 can protect DAergic neurons against MPTP- and MPP(+)-mediated damage. However, the effects of the toxins as well as of FGF-2 are partially dependent on culture conditions. Variations in the effectiveness of toxins and FGF-2 are not overtly related to the total numbers of neurons or astroglial cells, but may reflect culture type-dependent alterations of neuronal and glial metabolism.
利用胚胎第14天大鼠中脑的解离细胞培养物，研究了碱性成纤维细胞生长因子（FGF-2）对1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）和甲基吡啶离子（MPP+）损伤的多巴胺能（DAergic）黑质纹状体神经元的保护作用。细胞在不同的培养基中生长，并在不同的时间接受FGF-2（5 ng/ml）和/或毒素（5 μM），但在暴露于毒素之前，始终允许其分化3天。如果从培养基中省略马血清（HS）或牛血清白蛋白（BSA），则在7天时酪氨酸羟化酶（TH）免疫反应性细胞的存活仅受到MPTP的显著损害。FGF-2增加了TH免疫反应性细胞的数量，并且在任何培养条件下，MPTP都不会减弱这种增加。在含有HS和BSA的培养基中，MPTP显著降低了3H-DA的摄取，但在无蛋白培养基中则没有。仅在存在BSA的情况下，FGF-2才具有保护作用。MPP+比MPTP导致3H-DA摄取的降低更明显，并且除非培养物含有HS，否则添加FGF-2可部分逆转这种作用。神经丝蛋白（NF）是培养物中存在的神经元总数的间接测量指标，MPTP或MPP+并未使其显著降低，这证实了毒素对多巴胺能神经元的特异性，而多巴胺能神经元在这些培养物中仅占一小部分。与FGF-2广泛的靶神经元谱一致，该因子在任何培养条件下均显著增加NF含量。对胶质纤维酸性蛋白（GFAP）含量的定量分析显示，FGF-2具有刺激作用（2.5至4倍），并且与不存在HS相比，存在HS时其水平至少高出10倍。这些数据表明，FGF-2可以保护多巴胺能神经元免受MPTP和MPP+介导的损伤。然而，毒素以及FGF-2的作用部分取决于培养条件。毒素和FGF-2有效性的变化与神经元或星形胶质细胞的总数没有明显关系，但可能反映了培养类型依赖性的神经元和胶质细胞代谢改变。
