Van der Neut R, Bär P R, Sodaar P, Gispen W H
Research Laboratory Neurology, University of Utrecht, The Netherlands.
Peptides. 1988 Sep-Oct;9(5):1015-20. doi: 10.1016/0196-9781(88)90082-4.
Slices of foetal spinal cords in culture were used to establish possible trophic effects of alpha-melanocyte stimulating hormone (alpha-MSH) and a fragment of the adrenocorticotropic hormone (ACTH4-10) on the outgrowth of neurites from spinal neurons. The spinal cord slices were treated with peptides over a wide concentration range. Using monoclonal antibodies against (subunits of) neurofilament followed by immunofluorescence, we could show that the extension consisted mainly of axons. After 5 and 7 days, outgrowth was quantified with 2 different techniques, namely by visual scoring under phase contrast and by means of an ELISA for neurofilament protein. Both methods yielded the same dose-response profile. Both alpha-MSH and ACTH4-10 stimulated the formation of neurites in a dose-dependent manner, with a maximal stimulatory effect at 0.001-0.01 nM (ACTH4-10) or 0.1-1.0 nM (alpha-MSH). The maximal effect of the peptides was 30-40% compared to controls. We conclude that alpha-MSH and ACTH4-10 stimulate axonal outgrowth from foetal spinal cord slices in vitro in a dose-dependent way.
利用培养的胎儿脊髓切片来确定α-黑素细胞刺激素（α-MSH）和促肾上腺皮质激素片段（ACTH4-10）对脊髓神经元神经突生长可能产生的营养作用。在很宽的浓度范围内用这些肽处理脊髓切片。使用抗神经丝（亚基）的单克隆抗体，随后进行免疫荧光检测，我们能够证明这种延伸主要由轴突组成。在5天和7天后，用两种不同技术对生长情况进行定量，即相差显微镜下的视觉评分和神经丝蛋白的酶联免疫吸附测定法（ELISA）。两种方法得出相同的剂量反应曲线。α-MSH和ACTH4-10均以剂量依赖方式刺激神经突的形成，在0.001 - 0.01 nM（ACTH4-10）或0.1 - 1.0 nM（α-MSH）时具有最大刺激作用。与对照组相比，这些肽的最大作用为30% - 40%。我们得出结论，α-MSH和ACTH4-10在体外以剂量依赖方式刺激胎儿脊髓切片的轴突生长。
