Nishi R, Berg D K
J Neurosci. 1981 May;1(5):505-13. doi: 10.1523/JNEUROSCI.01-05-00505.1981.
Survival and development of chick ciliary ganglion neurons in vivo appear to depend on information from the embryonic eye structure that contains the postsynaptic targets of the neurons. We have tested embryonic eye extracts on ciliary ganglion neurons in dissociated cell culture for stimulation of growth and development. Control conditions were chosen that permitted the long term maintenance of the neurons in the absence of tissue extracts of conditioned medium. The conditions included coating the culture substratum with fibroblast material and increasing the K+ concentration in the culture medium to 25 mM. Neurons survived for at least 3 weeks in control conditions. Two major components were resolved in eye extracts that stimulated growth and development of the neurons above the basal levels obtained with control conditions. One component, with an apparent molecular weight of about 2 X 10(4) by gel filtration analysis, stimulated neuronal growth without increasing the levels of choline acetyltransferase activity per neuron. The second component, with an apparent molecular weight of about 5 X 10(4), increased development of choline acetyltransferase levels per neuron but had no effect on neuronal growth. Both components were effective in normal K+ as well as 25 mM K+. These components may represent mechanisms by which the postsynaptic target tissue acts in vivo to direct the growth and development of ciliary ganglion neurons.
鸡睫状神经节神经元在体内的存活和发育似乎依赖于来自胚胎眼结构的信息,该结构包含神经元的突触后靶标。我们在解离细胞培养物中的睫状神经节神经元上测试了胚胎眼提取物对生长和发育的刺激作用。选择了对照条件,使得神经元在没有条件培养基的组织提取物的情况下能够长期维持。这些条件包括用成纤维细胞材料包被培养底物,并将培养基中的钾离子浓度提高到25 mM。在对照条件下,神经元存活至少3周。在眼提取物中分离出两种主要成分,它们刺激神经元的生长和发育,使其高于对照条件下获得的基础水平。一种成分通过凝胶过滤分析,其表观分子量约为2×10⁴,刺激神经元生长但不增加每个神经元的胆碱乙酰转移酶活性水平。第二种成分,表观分子量约为5×10⁴,增加每个神经元的胆碱乙酰转移酶水平的发育,但对神经元生长没有影响。两种成分在正常钾离子浓度以及25 mM钾离子浓度下均有效。这些成分可能代表了突触后靶组织在体内指导睫状神经节神经元生长和发育的机制。