Xue Z G, Smith J
Institut d'Embryologie du CNRS, Nogent-sur-Marne, France.
J Neurosci. 1988 Mar;8(3):806-13. doi: 10.1523/JNEUROSCI.08-03-00806.1988.
During embryonic life, avian sensory ganglia contain cells with the potential to express, under appropriate experimental conditions, a number of properties characteristic of autonomic sympathetic neurons. Thus, cells capable of synthesizing noradrenaline (NA) from tyrosine differentiate when dorsal root ganglia (DRG) from 10-15 d embryonic quail are grown in culture (Xue et al., 1985a, b). In the present study, we show that cultures of DRG from 10 d embryos can take up 3H-NA by a high-affinity (Km = 1.0 microM), temperature-dependent process that can be inhibited by desmethylimipramine. By means of combined immunocytochemistry and autoradiography, it was demonstrated that the majority (70-80%) of the tyrosine hydroxylase (TH)-immunoreactive cells that developed in the cultures possessed a transport system for NA. Catecholamine (CA) uptake also occurred in a small, but relatively constant, number of TH-negative cells, but was absent from substance P-containing neurons. In contrast to TH, which appears only after 3-4 d in vitro, cells capable of taking up NA with high affinity were found in DRG cultures after only a few hours, and a small number (less than 0.5% of the total cell population) was detected in freshly removed, uncultured ganglia. Such cells did not react with antibodies directed against substance P or neurofilament proteins. We conclude that autonomic precursors are identifiable in a subset of non-neuronal DRG cells, prior to full expression of a noradrenergic phenotype, by their possession of a high-affinity uptake system for CA.
在胚胎期,禽类感觉神经节包含一些细胞,在适当的实验条件下,这些细胞有潜力表达自主交感神经元的一些特性。因此,当将10 - 15日龄胚胎鹌鹑的背根神经节(DRG)进行培养时,能够从酪氨酸合成去甲肾上腺素(NA)的细胞会发生分化(薛等人,1985a,b)。在本研究中,我们发现10日龄胚胎的DRG培养物能够通过一种高亲和力(Km = 1.0 microM)、依赖温度的过程摄取3H - NA,该过程可被去甲丙咪嗪抑制。通过免疫细胞化学和放射自显影相结合的方法,证明在培养物中发育的大多数(70 - 80%)酪氨酸羟化酶(TH)免疫反应阳性细胞拥有NA转运系统。儿茶酚胺(CA)摄取也发生在少量但相对恒定数量的TH阴性细胞中,但含P物质的神经元中不存在这种摄取。与仅在体外培养3 - 4天后才出现的TH不同,在DRG培养物中,仅培养数小时后就能发现具有高亲和力摄取NA能力的细胞,并且在刚取出的未培养神经节中检测到少量(占总细胞群体不到0.5%)这样的细胞。这些细胞不与针对P物质或神经丝蛋白的抗体发生反应。我们得出结论,在非神经元DRG细胞的一个亚群中,在去甲肾上腺素能表型完全表达之前,通过其拥有的高亲和力CA摄取系统可以识别出自主神经前体。