Carr V M, Simpson S B
J Comp Neurol. 1978 Dec 15;182(4):741-55. doi: 10.1002/cne.901820411.
Responses of chick embryo dorsal root ganglia to early wing bud amputation were examined histologically using tritiated thymidine (3H-TdR) and autoradiography to analyze proliferation and the Feulgen procedure to visualize degenerating cells. Right wing buds were amputated at stage 15 or 16. At 4.5 to 9.5 days of incubation embryos were given a 1-hour exposure to 3H-TdR and fixed. Feulgen-stained autoradiographs were examined for percentage of cells labelled (labelling index) or degenerating (degeneration index) in lateroventral (LV) and mediodorsal (MD) regions of brachial (G14-16) and nonbrachial (G12, 13, 17) ganglia. The earliest response to amputation was a highly significant increase in degeneration indices of LV and MD regions of ipsilateral brachial ganglia at 5.5 days. Significant brachial LV responses were observed throughout the remainder of the experimental period. Two peaks occur in this response: at 5.5 days, corresponding to the peak seen in normal nonbrachial ganglia, and at 8.5 days, having no counterpart in normal development. In brachial MD regions significant degenerative responses occur at most times examined. Significant responses also occur at 7.5 and 8.5 days in MD regions of nonbrachial ganglia. The presence of MD responses in our material indicates that maturation of at least some MD neurons occurs earlier than previously thought. Significant labelling responses occur in brachial LV regions from 7.5 days on. Because other studies (Carr and Simpson, '78a) show that this time is after the end of large-scale neuronal production, this labelling response must be nonneuronal in nature. We conclude that this response is a secondary response to amputation, consequent to the greatly increased cellular degeneration. Results of experiments involving addition of limb buds at the brachial level are also presented.
利用氚标记胸腺嘧啶核苷(³H-TdR)和放射自显影技术对鸡胚背根神经节对早期翅芽截肢的反应进行了组织学检查,以分析细胞增殖情况,并采用福尔根染色法观察退化细胞。在第15或16阶段切除右侧翅芽。在孵化4.5至9.5天时,给胚胎1小时的³H-TdR暴露时间,然后固定。对福尔根染色的放射自显影片进行检查,以确定臂神经节(G14 - 16)和非臂神经节(G12、13、17)的后腹侧(LV)和中背侧(MD)区域中标记细胞(标记指数)或退化细胞(退化指数)的百分比。截肢后的最早反应是在5.5天时,同侧臂神经节LV和MD区域的退化指数显著升高。在整个实验期的剩余时间里,均观察到明显的臂神经节LV反应。该反应出现两个峰值:一个在5.5天,与正常非臂神经节中出现的峰值相对应;另一个在8.5天,在正常发育中没有对应情况。在大多数检查时间点,臂神经节MD区域都出现明显的退化反应。非臂神经节MD区域在7.5天和8.5天也出现明显反应。我们的材料中存在MD反应表明,至少一些MD神经元的成熟比之前认为的要早。从7.5天开始,臂神经节LV区域出现明显的标记反应。由于其他研究(Carr和Simpson,1978a)表明这个时间是在大规模神经元生成结束之后,所以这种标记反应本质上一定是非神经元的。我们得出结论,这种反应是截肢后的次级反应,是细胞退化大幅增加的结果。还展示了在臂水平添加肢芽的实验结果。