NGF and anti-NGF: evidence against effects on fiber growth in locus coeruleus from cultures of perinatal CNS tissues.
NGF and anti-NGF: evidence against effects on fiber growth in locus coeruleus from cultures of perinatal CNS tissues.
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NGF 和抗 NGF：来自围产期 CNS 组织培养物对蓝斑纤维生长影响的证据。
DOI：
10.1159/000112451
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发表时间：
1979
影响因子：
2.9
通讯作者：
A. Seiger
中科院分区：
文献类型：
--
作者：
L. Olson;T. Ebendal;A. Seiger
本研究使用胶原凝胶组织培养技术检查蓝斑正在发育的去甲肾上腺素能神经元是否依赖于内源性神经生长因子（NGF）来产生神经纤维，以及外源性 NGF 是否刺激该核中的纤维生长。取自围产期大鼠脑的蓝藻用于三种培养实验：(1)从新生大鼠中制备蓝藻、顶叶大脑皮层和颈上神经节，并在同一培养皿的不同部分进行培养； (2)以相同方式培养的17日龄大鼠胎儿的蓝斑和顶叶大脑皮层，以及(3)与8日龄鸡胚的脊髓、交感神经节和睫状神经节共培养的17日龄大鼠胎儿的蓝斑。实验1和2在添加和不添加NGF和抗NGF的情况下进行，实验3在添加和不添加抗NGF的情况下进行。在 4 天内，每天通过暗视野和相差显微镜评估所有培养组织中的总纤维产量。然后通过 Falck-Hillarp 荧光组织化学研究了大鼠相同蓝斑和颈上神经节中肾上腺素能神经纤维的产生。胎鼠的蓝斑和大脑皮层在培养物中产生致密的纤维晕。新生大鼠的蓝斑产生的纤维相当少，新生大鼠的皮质仅产生很少的纤维。来自同一新生动物的颈上神经节不产生或几乎不产生纤维。正如在活体培养物和荧光显微镜中观察到的，添加 NGF 不能刺激蓝斑和大脑皮层的纤维生长。同样，添加抗 NGF 并不影响中枢神经系统区域的纤维产生。 NGF 对新生儿蓝斑和大脑皮层的阴性结果与在相同培养皿中培养的相同动物的颈上神经节所表现出的强烈、高度显着的纤维生长反应形成鲜明对比。第三个实验测试了组织培养物中的蓝斑是否含有或产生神经生长因子或三种鸡胚神经节中的任何一种。在这三个神经节中没有观察到任何反应。结论是，正在发育的蓝斑区不含有也不产生NGF，不依赖NGF产生纤维，并且不受外源性NGF刺激。
The present study examines whether the developing noradrenergic neurons of locus coeruleus depend on endogenous nerve growth factor (NGF) for nerve fiber production and if exogenous NGF stimulates fiber growth in this nucleus, using a collagen gel tissue culture technique. Lucus coeruleus from perinatal rat brain was used in three culture experiments: (1) lucus coeruleus, parietal cerebral cortex, and the superior cervical ganglion, prepared from newborn rats and cultured in different sectors of the same dishes; (2) locus coeruleus and parietal cerebral cortex from 17-day-old rat fetuses cultured in the same manner, and (3) locus coeruleus from 17-day-old rat fetuses co-cultured with spinal, sympathetic and ciliary ganglia from 8-day chick embryos. Experiments 1 and 2 were run with and without addition of NGF and anti-NGF, experiment 3 with and without anti-NGF. Total fiber production in all cultured tissues was evaluated daily by dark field and phase contrast microscopy during 4 days. Adrenergic nerve fiber production was then studied in the same locus coeruleus and superior cervical ganglia from the rats by Falck-Hillarp fluorescence histochemistry. Locus coeruleus and cortex cerebri from fetal rats produced dense fiber halos in culture. Locus coeruleus from newborn rats produced considerably less fibers, newborn cortex only few fibers. Superior cervical ganglia from the same newborn animals produced no or almost no fibers. Addition of NGF was not able to stimulate fiber growth in locus coeruleus nor in cortex cerebri as observed both in the living cultures and by fluorescence microscopy. Likewise, addition of anti-NGF did not affect fiber production in the CNS areas. The negative results with NGF on newborn locus coeruleus and cortex cerebri were in sharp contrast to the strong, highly significant fiber growth response demonstrated by the superior cervical ganglion from the same animals cultured in the same dishes. The third experiment tested whether locus coeruleus in tissue culture contained or produced nerve growth factor or any one of the three chick embryo ganglia. No response whatsoever in these three ganglia was observed. It is concluded that the developing locus coeruleus area does not contain or produce NGF, does not depend on NGF for fiber production, and is not stimulated by exogenous NGF.